HADHA, the alpha subunit of the mitochondrial trifunctional protein, is involved in long-chain fatty -induced autophagy in intestinal epithelial cells.

Genome-wide association studies have identified autophagy-related susceptibility genes for inflammatory bowel disease (); however, whether autophagy regulators can be utilized as therapeutic targets remains unclear. To identify novel microtubule-associated protein 1 light chain 3 (LC3)-interacting proteins in intestinal epithelial cells (IECs), we isolated primary IECs from green fluorescent protein (GFP)-LC3 mice. We performed immunoprecipitation with a GFP antibody and then analyzed co-immunoprecipitates by mass spectrometry. HADHA was identified as an LC3-interacting protein from primary IECs. The HADHA gene encodes the alpha subunit of the mitochondrial trifunctional protein. Given that HADHA catalyzes the last three steps of mitochondrial beta-oxidation of long-chain fatty acids, we investigated whether long-chain fatty acids induce autophagy in IECs. We found that induced autophagy in DLD-1, HT29, and HCT116\xa0cells. HADHA was expressed in not only the mitochondria but also the cytosol. LC3 puncta co-localized with HADHA, which were enhanced by stimulation. However, LC3 puncta did not co-localize with Tom20, suggesting that HADHA was induced to associate with LC3 puncta at sites other than the mitochondria. Thus, HADHA may have extra-mitochondrial functions. Furthermore, we found that induced cell death in IECs, which was accelerated by bafilomycin A and chloroquine. These findings suggested that -induced autophagy supports the survival of IECs. Taken together, these results suggested that HADHA is involved in long-chain fatty -induced autophagy in IECs, thus providing new insights into the pathology of and revealing novel therapeutic targets of .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: IBD

Lactobacillus johnsonii glycolipids, their structure and immunoreactivity with sera from inflammatory bowel disease patients.

Structural studies of the major glycolipids produced by two Lactobacillus johnsonii (LJ) strains, LJ 151 isolated from intestinal tract of healthy mice and LJ 142 isolated from mice with experimentally induced inflammatory bowel disease (), were performed. Two major glycolipids, GL1 and GL2, were present in lipid extracts from L.\xa0johnsonii 142 and 151 strains. Glycolipid GL1 has been identified as β-D-Glcp-(1→6)-α-D-Galp-(1→2)-α-D-Glcp-diglyceride and GL2 as α-D-Galp-(1→2)-α-D-Glcp-diglyceride. The main fatty residues identified by gas-liquid chromatography-mass spectrometry were , stearic and lactobacillic acids. Besides structural elucidation of the major glycolipids, the aim of this study was to determine the immunochemical properties of these glycolipids and to compare their immunoreactivity to that of polysaccharides obtained from the same strains. Sera from rabbits immunized with bacterial cells possessed much higher serological reactivity with polysaccharides than with glycolipids. Inversely, reactivity of the glycolipids with human sera from patients with was much higher than that determined for the polysaccharides, while reactivity of glycolipids with human sera from healthy individuals was much lower than one measured for the polysaccharides. Results indicate that glycoconjugates from Lactobacillus cell wall act as antigens and may represent new diagnostic biomarkers.© 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Keyword: IBD

Evidence for a new type of outer membrane lipid in oral spirochete Treponema denticola. Functioning permeation barrier without lipopolysaccharides.

A new class of outer membrane lipid (OML) was isolated from the oral spirochete Treponema denticola strain ATCC 33521 using a phenol/chloroform/light petroleum procedure normally applied for lipopolysaccharide extraction. In addition to chemical analysis, Fourier transform infrared (FTIR) spectroscopy was applied to compare the biophysical properties of OML with lipopolysaccharides (LPS) and lipoteichoic acids (LTA). Isolated OML fractions represent 1.4% of the total dry cell weight, are about 4 kDa in size, and contain 6% amino sugars, 8% neutral sugars, 14% phosphate, 35% carbazol-positive compounds, and 11% fatty acids (containing iso- and anteiso-fatty acyl chains). Rare for outer membrane lipids, OML contains no significant amount of 3-deoxy-D-manno-octulosonic acids, heptoses, and beta-hydroxy fatty acids. The fatty acyl chain composition, being similar to that of the cytoplasmic membrane, is quite heterogeneous with anteiso-pentadecanoic (12%), (51%), and iso- (19%) as the predominant fatty acids present. Findings of a glycerol-hexose unit and two glycerol-hexadecanoic fragments indicate a glycolipid membrane anchor typically found in LTA. There was also no evidence for the presence of a sphingosine-based lipid structure. The results of FTIR measurements strongly suggest that the reconstituted lipid forms normal bilayer structures (vesicles) expressing a high membrane state of order with a distinct phase transition as typical for isolated LPS. However, in contrast to LPS, OML of T. denticola has a lower Tm near 22 degreesC and a lower cooperativity of the phase transition. The results suggest a different kind of permeation barrier that is built up by this particular OML of T. denticola, which is quite different from LPS normally essential for Gram-negative bacteria.

Keyword: IBD

Docosahexaenoyl serotonin emerges as most potent inhibitor of IL-17 and CCL-20 released by blood mononuclear cells from a series of N-acyl serotonins identified in human intestinal tissue.

Fatty amides (FAAs), conjugates of fatty acids with ethanolamine, mono-amine neurotransmitters or amino acids are a class of molecules that display diverse functional roles in different cells and tissues. Recently we reported that one of the serotonin-fatty conjugates, docosahexaenoyl serotonin (DHA-5-HT), previously found in gut tissue of mouse and pig, attenuates the IL-23-IL-17 signaling axis in LPS-stimulated mice macrophages. However, its presence and effects in humans remained to be elucidated. Here, we report for the first time its identification in human intestinal (colon) tissue, along with a series of related N-acyl serotonins. Furthermore, we tested these fatty conjugates for their ability to inhibit the release of IL-17 and CCL-20 by stimulated human peripheral blood mononuclear cells (PBMCs). Serotonin conjugates with (PA-5-HT), stearic (SA-5-HT) and oleic (OA-5-HT) were detected in higher levels than arachidonoyl serotonin (AA-5-HT) and DHA-5-HT, while eicosapentaenoyl serotonin (EPA-5-HT) could not be quantified. Among these, DHA-5-HT was the most potent in inhibiting IL-17 and CCL-20, typical Th17 pro-inflammatory mediators, by Concanavalin A (ConA)-stimulated human PBMCs. These results underline the idea that DHA-5-HT is a gut-specific endogenously produced mediator with the capacity to modulate the IL-17/Th17 signaling response. Our findings may be of relevance in relation to intestinal inflammatory diseases like Crohn's disease and Ulcerative colitis.Copyright © 2017. Published by Elsevier B.V.

Keyword: IBD

Enhancing growth and lipid production of marine microalgae for biodiesel production via the use of different LED wavelengths.

Wavelength of light is a crucial factor which renders microalgae as the potential biodiesel. In this study, Tetraselmis sp. and Nannochloropsis sp. as famous targets were selected. The effect of different light wavelengths on growth rate and lipid production was studied. Microalgae were cultivated for 14 days as under blue, red, red-blue LED and white fluorescent light. The growth rate of microalgae was analyzed by spectrophotometer and cell counting while oil production under improved Nile red method. Optical density result showed the microalgae exhibited better growth curve under blue wavelength. Besides, Tetraselmis sp. and Nannochloropsis sp. under blue wavelength showed the higher growth rate (1.47 and 1.64 day(-1)) and oil production (102.954 and 702.366 a.u.). Gas chromatography analysis also showed that and stearic which were compulsory components for biodiesel contribute around 49-51% of total FAME from Nannochloropsis sp. and 81-83% of total FAME from Tetraselmis sp.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: IBD

Comprehensive genetic study of fatty acids helps explain the role of noncoding inflammatory bowel disease associated SNPs and fatty metabolism in disease pathogenesis.

Fatty acids and their derivatives play an important role in inflammation. Diet and genetics influence fatty profiles. Abnormalities of fatty profiles have been observed in inflammatory bowel diseases (), a group of complex diseases defined by chronic gastrointestinal inflammation. associated fatty profile abnormalities were observed independently of nutritional status or disease activity, suggesting a common genetic background. However, no study so far has attempted to look for overlap between loci and fatty associated loci or investigate the genetics of fatty profiles in . To this end, we conducted a comprehensive genetic study of fatty profiles in using iCHIP, a custom microarray platform designed for deep sequencing of immune-mediated disease associated loci. This study identifies 10 loci associated with fatty profiles in . The most significant associations were a locus near CBS (p\u202f=\u202f7.62\u202f×\u202f10) and a locus in LRRK2 (p\u202f=\u202f1.4\u202f×\u202f10). Of note, this study replicates the FADS gene cluster locus, previously associated with both fatty profiles and pathogenesis. Furthermore, we identify 18 carbon chain trans-fatty acids (p\u202f=\u202f1.12\u202f×\u202f10), total trans-fatty acids (p\u202f=\u202f4.49\u202f×\u202f10), (p\u202f=\u202f5.85\u202f×\u202f10) and arachidonic (p\u202f=\u202f8.58\u202f×\u202f10) as significantly associated with pathogenesis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: IBD

N-Acylethanolamine-hydrolyzing amidase inhibition increases colon N-palmitoylethanolamine levels and counteracts murine colitis.

N-Palmitoylethanolamine or palmitoylethanolamide (PEA) is an anti-inflammatory compound that was recently shown to exert peroxisome proliferator-activated receptor-α-dependent beneficial effects on colon inflammation. The actions of PEA are terminated following hydrolysis by 2 enzymes: fatty amide hydrolase (FAAH), and the less-studied N-acylethanolamine-hydrolyzing amidase (NAAA). This study aims to investigate the effects of inhibiting the enzymes responsible for PEA hydrolysis in colon inflammation in order to propose a potential therapeutic target for inflammatory bowel diseases (IBDs). Two murine models of were used to assess the effects of NAAA inhibition, FAAH inhibition, and PEA on macroscopic signs of colon inflammation, macrophage/neutrophil infiltration, and the expression of proinflammatory mediators in the colon, as well as on the colitis-related systemic inflammation. NAAA inhibition increases PEA levels in the colon and reduces colon inflammation and systemic inflammation, similarly to PEA. FAAH inhibition, however, does not increase PEA levels in the colon and does not affect the macroscopic signs of colon inflammation or immune cell infiltration. This is the first report of an anti-inflammatory effect of a systemically administered NAAA inhibitor. Because NAAA is the enzyme responsible for the control of PEA levels in the colon, we put forth this enzyme as a potential therapeutic target in chronic inflammation in general and in particular.© FASEB.

Keyword: IBD

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[Protective effects of total saponins of Panax japonicas on HepG2 cell apoptosis induced with ].

This study aimed to investigate the molecular mechanism and protective effect of total saponins of Panax japonicas (TSPJ) on HepG2 cells apoptosis induced by (PA).The HepG2 cells were cultured , and divided into five groups: the control group, the model group, the high-dose group (50 mg·L⁻¹), the middle-dose group (25 mg·L⁻¹) and the low-dose group (12.5 mg·L⁻¹).The cells of the five groups were cultured continuously for 24 hours. The cell viability was measured with MTT. HepG2 cells apoptosis was detected by Hoechest staining and Annexin V-FITC/PI staining. The protein expressions of BCL-2, CHOP and TLR4 were measured with western blotting and flow cytometry analysis. The mRNA expressions of TNF-α, IL-1β, BCL-2, CHOP and GAPDH were measured with RT-PCR. The results suggested that compared with the control group, the number of HepG2 cells of the model group were reduced significantly (<0.01), while the number of apoptotic HepG2 cells were increased. Compared with the model group, the number of HepG2 cells of the high-dose group and the middle-dose group were increased significantly (<0.01), whereas the number of apoptotic HepG2 cells were reduced. Compared with the control group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the model group were significantly up-regulated (<0.01), while BCL-2 protein and mRNA expressions in the model group were significantly decreased (<0.01). Compared with the model group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the high-dose group were significantly decreased (<0.01), while BCL-2 protein and mRNA expressions in the high-dose group were significantly up-regulated (<0.01).In conclusion, TSPJ can reduce inflammation and apoptosis induced by , with a certain protective effect on liver cells.Copyright© by the Chinese Pharmaceutical Association.

Keyword: NASH

Inhibitory Effect of Gardenoside on Free Fatty -Induced Steatosis in HepG2 Hepatocytes.

Gardenoside is one of the most important effective extractions of a herb for its hepatoprotective properties. The aim of this study was to address the mechanism of Gardenoside on HepG2 cellular steatosis induced by free fatty acids (FFAs). The model of HepG2 steatosis was duplicated by oleic and at the proportion of 2:1 (FFAs mixture) for 24 h, then lipid toxicity was induced. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were used to detect cell viability and Oil Red O staining method was used to judge the lipid accumulation respectively. Inflammatory cytokines TNF-α, IL-1β, IL-6 and intracellular NFκB were measured after 24 h. The steatosis was significantly decreased after Gardenoside treatment without cytotoxicity. TNF-α, IL-1β, IL-6 were modulated to HepG2 cells by treatment of Gardenoside. In the meantime, the activation of NFκB was inhibited by Gardenoside. Gardenoside has a protective effect on FFA-induced cellular steatosis in HepG2 cells which indicates that Gardenoside might be a potential therapeutic herb against by suppressed supernatant inflammatory cytokine production and intracellular NFkB activity.

Keyword: NASH

Dual roles of ULK1 (unc-51 like autophagy activating kinase 1) in cytoprotection against lipotoxicity.

Saturated fatty (SFA)-induced lipotoxicity is caused by the accumulation of reactive oxygen species (ROS), which is associated with damaged mitochondria. Moreover, lipotoxicity is crucial for the progression of nonalcoholic steatohepatitis (). Autophagy is required for the clearance of protein aggregates or damaged mitochondria to maintain cellular metabolic homeostasis. The NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2)-KEAP1 (kelch like ECH associated protein 1) pathway is essential for the elimination of ROS. ULK1 (unc-51 like autophagy activating kinase 1; yeast Atg1) is involved in the initiation of autophagy; however, its role in lipotoxicity-induced cell death in hepatocytes and mouse liver has not been elucidated. We now show that ULK1 potentiates the interaction between KEAP1 and the autophagy adaptor protein SQSTM1/p62, thereby mediating NFE2L2 activation in a manner requiring SQSTM1-dependent autophagic KEAP1 degradation. Furthermore, ULK1 is required for the autophagic removal of damaged mitochondria and to enhance binding between SQSTM1 and PINK1 (PTEN induced kinase 1). This study demonstrates the molecular mechanisms underlying the cytoprotective role of ULK1 against lipotoxicity. Thus, ULK1 could represent a potential therapeutic target for the treatment of . Abbreviations: ACTB: actin beta; CM-HDCFDA:5-(and-6)-chloromethyl-2\',7\'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; LPS: lipopolysaccharides; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAPK8/JNK: mitogen-activated protein kinase 8; MEF: mouse embryonic fibroblast; MFN1: mitofusin 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; : nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: ; PARP: poly (ADP-ribose) polymerase 1; PINK1: PTEN induced kinase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PRKC/PKC: protein kinase C; RBX1: ring-box 1; ROS: reactive oxygen species; SFA: saturated fatty ; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; TUBA: tubulin alpha; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase 1.

Keyword: NASH

A novel palmitoyl acyl transferase controls surface protein palmitoylation and cytotoxicity in Giardia lamblia.

The intestinal protozoan parasite Giardia lamblia undergoes surface antigenic variation whereby one of a family of structurally related variant-specific surface proteins (VSPs) is replaced in a regulated process by another antigenically distinct VSP. All VSPs are type I membrane proteins that have a conserved hydrophobic sequence terminated by the invariant hydrophilic amino acids, CRGKA. Using transfected Giardia constitutively expressing HA-tagged VSPH7 and incubated with radioactive [3H]palmitate, we demonstrate that the palmitate is attached to the Cys in the conserved CRGKA tail. Surface location of mutant VSPs lacking either the CRGKA tail or its Cys is identical to that of wild-type VSPH7 but non-palmitoylated mutants fail to undergo complement-independent antibody specific cytotoxicity. In addition, membrane localization of non-palmitoylated mutant VSPH7 changes from a pattern similar to rafts to non-rafts. Palmitoyl transferases (PAT), responsible for protein palmitoylation in other organisms, often possess a cysteine-rich domain containing a conserved DHHC motif (DHHC-CRD). An open reading frame corresponding to a putative 50 kDa Giardia PAT (gPAT) containing a DHHC-CRD motif was found in the Giardia genome database. Expression of epitope-tagged gPAT using a tetracycline inducible vector localized gPAT to the plasma membrane, a pattern similar to that of VSPs. Transfection with gPAT antisense producing vectors inhibits gPAT expression and palmitoylation of VSPs in vitro confirming the function of gPAT. These results show that VSPs are palmitoylated at the cysteine within the conserved tail by gPAT and indicate an essential function of palmitoylation in control of VSP-mediated signalling and processing.

Keyword: NASH

Exosomes derived from -treated hepatocytes induce fibrotic activation of hepatic stellate cells.

Non-alcoholic fatty liver disease (NAFLD) is a dominant cause of chronic liver disease, but the exact mechanism of progression from simple steatosis to nonalcoholic steatohepatitis () remains unknown. Here, we investigated the role of exosomes in NAFLD progression. Exosomes were isolated from a human hepatoma cell line treated with (PA) and their miRNA profiles examined by microarray. The human hepatic stellate cell (HSC) line (LX-2) was then treated with exosome isolated from hepatocytes. Compared with controls, PA-treated hepatocytes displayed significantly increased CD36 and exosome production. The microarray analysis showed there to be distinctive miRNA expression patterns between exosomes from vehicle- and PA-treated hepatocytes. When LX-2 cells were cultured with exosomes from PA-treated hepatocytes, the expression of genes related to the development of fibrosis were significantly amplified compared to those treated with exosomes from vehicle-treated hepatocytes. In conclusion, PA treatment enhanced the production of exosomes in these hepatocytes and changed their exosomal miRNA profile. Moreover, exosomes derived from PA-treated hepatocytes caused an increase in the expression levels of fibrotic genes in HSCs. Therefore, exosomes may have important roles in the crosstalk between hepatocytes and HSCs in the progression from simple steatosis to .

Keyword: NASH

Intracellular and extracellular miRNome deregulation in cellular models of NAFLD or : Clinical implications.

Nonalcoholic fatty liver disease (NAFLD) represents the most common chronic liver disease in industrialized countries. NAFLD has the potential to progress through the inflammatory phase of nonalcoholic steatohepatitis () to fibrosis, cirrhosis, and hepatocellular carcinoma. Identifying patients at risk for this transition is a relevant clinical challenge. The complexity of these phenotypes in\xa0vivo made necessary the development of in\xa0vitro models in order to dissect the molecular signalling affected in NAFLD and , but also to identify potential circulating biomarkers.We profiled the expression of 754 cellular and medium-secreted human miRNAs in HepG2 cells after lipotoxic (Palmitate, model of ) or not-lipotoxic stimuli (Oleate-Palmitate, model of NAFLD). Results were validated through Single TaqMan assays. We performed computational analysis of miRNA targets and pathways. Oleate-palmitate treatment induced a variation of 2.8% and 10% of total miRNAs in cells and medium, respectively; palmitate treatment caused 10% and 19% intracellular and extracellular miRNA deregulation, respectively. We validated miR-126, miR-150, miR-223, miR-483-3p, miR-1226*, and miR-1290 deregulation. Through computational analysis, we observed that targets of both intracellular and extracellular DE miRNAs were involved in processes associated with the onset and progression of NAFLD and , such as fatty metabolism, apoptosis and inflammation.These data would be useful to elucidate the role of miRNAs in the pathogenesis and progression of the NAFLD spectrum, but they also allow the identification of novel potential biomarkers for differential diagnosis to be tested in\xa0vivo.Copyright © 2016 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

Keyword: NASH

PKC{delta} is activated in a dietary model of steatohepatitis and regulates endoplasmic reticulum stress and cell death.

Hepatic steatosis can progress to the clinical condition of non-alcoholic steatohepatitis (), which is a precursor of more serious liver diseases. The novel PKC isoforms δ and ε are activated by lipid metabolites and have been implicated in lipid-induced hepatic disease. Using a methionine- and choline-deficient (MCD) dietary model of , we addressed the question of whether hepatic PKCδ and PKCε are activated. With progression from steatosis to steatohepatitis, there was activation and increased PKCδ protein content coincident with hepatic endoplasmic reticulum (ER) stress parameters. To examine whether similar changes could be induced in vitro, McA-RH 7777 (McA) hepatoma cells were used. We observed that McA cells stored triglyceride and released alanine aminotransferase (ALT) when treated with MCD medium in the presence of fatty acids. Further, MCD medium with , but not oleic or linoleic acids, maximally activated PKCδ and stimulated ER stress. In PKCδ knockdown McA cells, MCD/fatty medium-induced ALT release and ER stress induction were completely blocked, but triglyceride storage was not. In addition, a reduction in the uptake of propidium iodide and the number of apoptotic nuclei and a significant increase in cell viability and DNA content were observed in PKCδ knockdown McA cells incubated in MCD medium with . Our studies show that PKCδ activation and protein levels are elevated in an animal model of steatohepatitis, which was recapitulated in a cell model, supporting the conclusion that PKCδ plays a role in ALT release, the ER stress signal, and cell death.

Keyword: NASH

Hepatic TLR4 signaling in obese NAFLD.

Nonalcoholic fatty liver disease occurs frequently in the setting of metabolic syndrome, but the factors leading to nonalcoholic steatohepatitis () are not fully understood. This study investigated Toll-like receptor 4 (TLR4) signaling in human liver with the goal of delineating whether activation of this pathway segregates those with nonalcoholic fatty liver from those with . Experiments were performed using liver biopsy tissue obtained from class III obese subjects undergoing bariatric surgery, and extended to an immortalized human hepatocyte HepaRG cell line and primary human hepatocytes. The bacterial endotoxin lipopolysaccharide (LPS) and total free fatty levels were significantly increased in plasma of patients. TLR4 mRNA levels were significantly increased in subjects with compared with NAFL as was interferon regulatory factor (IRF) 3 in the myeloid differentiation factor 88-independent signaling pathway. In HepaRG cells, nuclear factor-κB (NF-κB) nuclear translocation and functional activity increased following treatment with the fatty , palmitate, and following exposure to LPS compared with hepatocytes stimulated with a lipogenic treatment that induced de novo lipogenesis. Palmitate and LPS induction of NF-κB activity was partially attenuated by chemical- or small-interfering RNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was upregulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-κB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition. In conclusion, TLR4 expression is upregulated in a large cohort of patients, compared with those with NAFL, and this occurs within the setting of increased LPS and fatty acids.Copyright © 2015 the American Physiological Society.

Keyword: NASH

The NOX1 isoform of NADPH oxidase is involved in dysfunction of liver sinusoids in nonalcoholic fatty liver disease.

The increased production of reactive oxygen species (ROS) has been postulated to play a key role in the progression of nonalcoholic fatty liver disease (NAFLD). However, the source of ROS and mechanisms underlying the development of NAFLD have yet to be established. We observed a significant up-regulation of a minor isoform of NADPH oxidase, NOX1, in the liver of nonalcoholic steatohepatitis () patients as well as of mice fed a high-fat and high-cholesterol (HFC) diet for 8 weeks. In mice deficient in Nox1 (Nox1KO), increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 demonstrated in HFC diet-fed wild-type mice (WT) were significantly attenuated. Concomitantly, increased protein nitrotyrosine adducts, a marker of peroxynitrite-induced injury detected in hepatic sinusoids of WT, were significantly suppressed in Nox1KO. The expression of NOX1 mRNA was much higher in the fractions of enriched liver sinusoidal endothelial cells (LSECs) than in those of hepatocytes. In primary cultured LSECs, (PA) up-regulated the mRNA level of NOX1, but not of NOX2 or NOX4. The production of nitric oxide by LSECs was significantly attenuated by PA-treatment in WT but not in Nox1KO. When the in vitro relaxation of TWNT1, a cell line that originated from hepatic stellate cells, was assessed by the gel contraction assay, the relaxation of stellate cells induced by LSECs was attenuated by PA treatment. In contrast, the relaxation effect of LSECs was preserved in cells isolated from Nox1KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impaired hepatic microcirculation through the reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the progression of NAFLD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: NASH

Interleukin-17 exacerbates hepatic steatosis and inflammation in non-alcoholic fatty liver disease.

Mechanisms associated with the progression of simple steatosis to non-alcoholic fatty liver disease (NAFLD) remain undefined. Regulatory T cells (T(regs)) play a critical role in regulating inflammatory processes in non-alcoholic steatohepatitis () and because T helper type 17 (Th17) functionally oppose T(reg)-mediated responses, this study focused on characterizing the role of Th17 cells using a NAFLD mouse model. C57BL/6 mice were fed either a normal diet (ND) or high fat (HF) diet for 8 weeks. Mice in the HF group had a significantly higher frequency of liver Th17 cells compared to ND-fed mice. Neutralization of interleukin (IL)-17 in HF mice ameliorated lipopolysaccharide (LPS)-induced liver injury reflected by decreased serum alanine aminotransferase (ALT) levels and reduced inflammatory cell infiltrates in the liver. In vitro, HepG2 cells cultured in the presence of free fatty acids (FFA; oleic and ) for 24 h and IL-17 developed steatosis via insulin-signalling pathway interference. IL-17 and FFAs synergized to induce IL-6 production by HepG2 cells and murine primary hepatocytes which, in combination with transforming growth factor (TGF-β), expanded Th17 cells. It is likely that a similar process occurs in patients, as there were significant levels of IL-17(+) cell infiltrates in patient livers. The hepatic expression of Th17 cell-related genes [retinoid-related orphan receptor gamma (ROR)γt, IL-17, IL-21 and IL-23] was also increased significantly in patients compared to healthy controls. Th17 cells and IL-17 were associated with hepatic steatosis and proinflammatory response in NAFLD and facilitated the transition from simple steatosis to steatohepatitis. Strategies designed to alter the balance between Th17 cells and T(regs) should be explored as a means of preventing progression to and advanced liver diseases in NAFLD patients.© 2011 The Authors. Clinical and Experimental Immunology © 2011 British Society for Immunology.

Keyword: NASH

Macrophage Stimulating Protein Enhances Hepatic Inflammation in a Model.

Non-alcoholic steatohepatitis () is a common liver disease characterized by hepatic lipid accumulation (steatosis) and inflammation. Currently, therapeutic options are poor and the long-term burden to society is constantly increasing. Previously, macrophage stimulating protein (MSP)-a serum protein mainly secreted by liver-was shown to inhibit oxidized low-density lipoprotein (OxLDL)/lipopolysaccharides (LPS)-induced inflammation in mouse macrophages. Additionally, MSP could reduce (PA)-induced lipid accumulation and lipogenesis in the HepG2 cell line. Altogether, these data suggest MSP as a suppressor for metabolic inflammation. However, so far the potential of MSP to be used as a treatment for was not investigated. We hypothesized that MSP reduces lipid accumulation and hepatic inflammation. To investigate the effects of MSP in the early stage of , low-density lipoprotein receptor (Ldlr-/-) mice were fed either a regular chow or a high fat, high cholesterol (HFC) diet for 7 days. Recombinant MSP or saline (control) was administrated to the mice by utilizing subcutaneously-implanted osmotic mini-pumps for the last 4 days. As expected, mice fed an HFC diet showed increased plasma and hepatic lipid accumulation, as well as enhanced hepatic inflammation, compared with chow-fed controls. Upon MSP administration, the rise in cholesterol and triglyceride levels after an HFC diet remained unaltered. Surprisingly, while hepatic macrophage and neutrophil infiltration was similar between the groups, MSP-treated mice showed increased gene expression of pro-inflammatory and pro-apoptotic mediators in the liver, compared with saline-treated controls. Contrary to our expectations, MSP did not ameliorate . Observed changes in inflammatory gene expression suggest that further research is needed to clarify the long-term effects of MSP.

Keyword: NASH

ASMase regulates autophagy and lysosomal membrane permeabilization and its inhibition prevents early stage non-alcoholic steatohepatitis.

sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (). However, the contribution of ASMase to is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced .Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase(-/-) mice fed a HFD. The impact of pharmacological ASMase inhibition on was analyzed in wild type mice fed a HFD.ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase(-/-) mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase(-/-) mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase(-/-) mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase(+/+) mice but not in ASMase(-/-) mice. ASMase(-/-) PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methyl-serine dodecylamide hydrochloride or , effects that were reversed by decreasing cholesterol levels by oxysterol 25-hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage .These findings underscore a critical role for ASMase in diet-induced and suggest the potential of amitriptyline as a treatment for patients with .Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: NASH

Fatty activates NLRP3 inflammasomes in mouse Kupffer cells through mitochondrial DNA release.

Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease in many developed and developing countries worldwide. It has been well established that the chronic sterile inflammation caused by the NLRP3 inflammasome is closely related to NAFLD development. Kupffer cells (KCs) are involved in the pathogenesis of various liver diseases. We used methionine choline-deficient diets to establish a mouse nonalcoholic steatohepatitis () model. The expression and formation of the NLRP3 inflammasome in the KCs from the mouse and cell models were determined by Western blotting and co-immunoprecipitation. Evidence of mitochondrial DNA (mtDNA) release was determined by live cell labeling and imaging. KCs and the NLRP3 inflammasome exerted proinflammatory effects on the development and progression of through secretion of the proinflammatory cytokine IL-1β. NLRP3, ASC and Caspase-1 protein expression levels in KCs from mouse livers were significantly higher than those in KCs from NLRP3 mice, and the number of NLRP3 inflammasome protein complexes was significantly higher in KCs from mouse livers, whereas these protein complexes could not be formed in NLRP3 mice. In in vitro experiments, (PA) decreased the mitochondrial membrane potential and subsequently induced mtDNA release from the mitochondria to the cytoplasm. NLRP3 inflammasome expression was substantially increased, and mtDNA-NLRP3 inflammasome complexes formed upon PA stimulation. Our data suggest that mtDNA released from mitochondria during PA stimulation causes NLRP3 inflammasome activation, providing a missing link between NLRP3 inflammasome activation and development, via binding of cytosolic mtDNA to the NLRP3 inflammasome.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: NASH

Impaired autophagic flux is associated with increased endoplasmic reticulum stress during the development of NAFLD.

The pathogenic mechanisms underlying the progression of non-alcoholic fatty liver disease (NAFLD) are not fully understood. In this study, we aimed to assess the relationship between endoplasmic reticulum (ER) stress and autophagy in human and mouse hepatocytes during NAFLD. ER stress and autophagy markers were analyzed in livers from patients with biopsy-proven non-alcoholic steatosis (NAS) or non-alcoholic steatohepatitis () compared with livers from subjects with histologically normal liver, in livers from mice fed with chow diet (CHD) compared with mice fed with high fat diet (HFD) or methionine-choline-deficient (MCD) diet and in primary and Huh7 human hepatocytes loaded with (PA). In patients, significant increases in hepatic messenger RNA levels of markers of ER stress (activating transcription factor 4 (ATF4), glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP)) and autophagy (BCN1) were found compared with NAS patients. Likewise, protein levels of GRP78, CHOP and p62/SQSTM1 (p62) autophagic substrate were significantly elevated in compared with NAS patients. In livers from mice fed with HFD or MCD, ER stress-mediated signaling was parallel to the blockade of the autophagic flux assessed by increases in p62, microtubule-associated protein 2 light chain 3 (LC3-II)/LC3-I ratio and accumulation of autophagosomes compared with CHD fed mice. In Huh7 hepatic cells, treatment with PA for 8 h triggered activation of both unfolding protein response and the autophagic flux. Conversely, prolonged treatment with PA (24 h) induced ER stress and cell death together with a blockade of the autophagic flux. Under these conditions, cotreatment with rapamycin or CHOP silencing ameliorated these effects and decreased apoptosis. Our results demonstrated that the autophagic flux is impaired in the liver from both NAFLD patients and murine models of NAFLD, as well as in lipid-overloaded human hepatocytes, and it could be due to elevated ER stress leading to apoptosis. Consequently, therapies aimed to restore the autophagic flux might attenuate or prevent the progression of NAFLD.

Keyword: NASH

Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis.

Mitochondrial dysfunction plays a crucial role in the development of non-alcoholic steatohepatitis (). However, the regulator of mitochondrial dysfunction in the pathogenesis of is still largely unclear. CXCR3 is an essential pro-inflammatory factor in chronic liver diseases. We explored the significance of CXCR3 in regulating mitochondrial function during development in animal models and cultured hepatocytes.The effects of CXCR3 on mitochondrial function were evaluated by genetic knockout or pharmacological inhibition in mouse models and . The ultrastructural changes of mitochondria were assessed by transmission electron microscopy (TEM). Hepatic levels of mitochondrial reactive oxygen species (ROS), DNA damage, membrane potential and ATP were examined.CXCR3 ablation by genetic knockout or pharmacological inhibition in mice protected against development by influencing mitochondrial function. Similarly, depletion of CXCR3 reduced steatohepatitis injury in cultured hepatocytes. TEM analysis revealed that liver mitochondrial integrity was much improved in CXCR3 knockout (CXCR3) compared to wildtype (WT) mice. In agreement with this, impaired mitochondrial function was pronounced in WT mice compared to CXCR3 mice, evidenced by increased protein expression of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and decreased protein expression of mitofusin-1 (MFN1). Mitochondrial dysfunction was induced in AML-12 hepatocytes by methionine and choline deficient medium and in HepG2 cells by . The impaired mitochondrial function in both cell lines was evidenced by reduced membrane potential and ATP content, and by increased mitochondrial ROS accumulation and DNA damage. However, CXCR3 knockdown by siCXCR3 significantly diminished the mitochondrial dysfunction in both AML-12 and HepG2 hepatocytes. In addition, inhibition of CXCR3 by CXCR3 specific antagonists SCH546738 and AMG487 restored mitochondrial function and inhibited mitochondrial-dependent apoptosis in the liver of WT mice fed with methionine and choline deficient diet.CXCR3 induces mitochondrial dysfunction, which contributes to the pathogenesis of steatohepatitis. Pharmacologic blockade of CXCR3 prevents mitochondrial dysfunction and restores the severity of steatohepatitis, indicating a potential clinical impact for controlling the disease.

Keyword: NASH

Oxidation of hepatic carnitine palmitoyl transferase-I (CPT-I) impairs fatty beta-oxidation in rats fed a methionine-choline deficient diet.

There is growing evidence that mitochondrial dysfunction, and more specifically fatty β-oxidation impairment, is involved in the pathophysiology of non-alcoholic steatohepatitis (). The goal of the present study was to achieve more understanding on the modification/s of carnitinepalmitoyltransferase-I (CPT-I), the rate-limiting enzyme of the mitochondrial fatty β-oxidation, during steatohepatitis. A high fat/methionine-choline deficient (MCD) diet, administered for 4 weeks, was used to induce in rats.We demonstrated that CPT-I activity decreased, to the same extent, both in isolated liver mitochondria and in digitonin-permeabilized hepatocytes from MCD-diet fed rats.At the same time, the rate of total fatty oxidation to CO(2) and ketone bodies, measured in isolated hepatocytes, was significantly lowered in treated animals when compared to controls. Finally, an increase in CPT-I mRNA abundance and protein content, together with a high level of CPT-I protein oxidation was observed in treated rats. A posttranslational modification of rat CPT-I during steatohepatitis has been here discussed.

Keyword: NASH

Anti-lipoapoptotic effect of Artemisia capillaris extract on free fatty acids-induced HepG2 cells.

Artemisia capillaris (AC) has been recognized as one of the promising candidates for hepatoprotective, hypoglycemic, hypolipidemic, antiobesitic and anti-inflammatory therapeutic effectiveness. This study evaluated the inherent mechanism and anti-apoptotic activity of 30% ethanol extract of AC (AC extract) 100 μg/ml on free fatty acids (FFAs)-induced HepG2 cellular steatosis and lipoapoptosis.Hepatic steatosis was induced by culturing HepG2 cells with a FFAs mixture (oleic and at the proportion of 2:1) for 24 h, thus ultimately giving rise to lipoapoptosis. Cell viability and lipid accumulation were detected by MTT assay and Oil Red O staining method respectively and Caspase-3, -9, Bax, Bcl-2, p-JNK and PUMA were measured for lipoapoptosis after 24 hours.AC extract significantly improved the FFAs-induced steatosis without cytotoxicity and Caspase-3, -9, Bax and Bcl-2 were modulated profitably to HepG2 cells after AC treatment. In addition, AC extract inhibited the activation of c-Jun NH2 terminal kinase (JNK) and PUMA, which mechanism is related to non-alcoholic steatohepatitis ().Combined together, AC extract exerted an obvious hypolipidemic and anti-apoptotic effect, indicating that AC extract might have potential therapeutic herb against .

Keyword: NASH

The phytochemical polydatin ameliorates non-alcoholic steatohepatitis by restoring lysosomal function and autophagic flux.

Impaired autophagic degradation of intracellular lipids is causally linked to the development of non-alcoholic steatohepatitis (). Pharmacological agents that can restore hepatic autophagic flux could therefore have therapeutic potentials for this increasingly prevalent disease. Herein, we investigated the effects of polydatin, a natural precursor of resveratrol, in a murine nutritional model of and a cell line model of steatosis. Results showed that oral administration of polydatin protected against hepatic lipid accumulation and alleviated inflammation and hepatocyte damage in db/db mice fed methionine-choline deficient diet. Polydatin also alleviated -induced lipid accumulation in cultured hepatocytes. In both models, polydatin restored lysosomal function and autophagic flux that were impaired by or steatosis. Mechanistically, polydatin inhibited mTOR signalling and up-regulated the expression and activity of TFEB, a known master regulator of lysosomal function. In conclusion, polydatin ameliorated through restoring autophagic flux. The polydatin-regulated autophagy was associated with inhibition of mTOR pathway and restoration of lysosomal function by TFEB. Our study provided affirmative preclinical evidence to inform future clinical trials for examining the potential anti- effect of polydatin in humans.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: NASH

Effects of on TNF-α-induced cytotoxicity in SK-Hep-1 cells.

Non-alcoholic steatohepatitis () is an increasingly common cause of chronic liver disease; however, no specific pharmacologic therapy has been shown to be effective in its treatment. The present study was designed to develop an experimental cell culture model of using four kinds of fatty acids - (PA), stearic (SA), linoleic (LA), and oleic (OA) - and TNF-α, according to the "two-hit" hypothesis. The saturated fatty acids PA and SA are more cytotoxic than the unsaturated fatty acids OA and LA. Cellular lipid accumulation without cytotoxicity was more easily induced with the unsaturated fatty acids than with the saturated fatty acids. PA augmented TNF-α-induced cytotoxicity, while the unsaturated fatty acids attenuated TNF-α-induced cytotoxicity. In a mechanistic study, PA enhanced TNF-α-mediated apoptosis in the absence of oxidative stress, as determined by measuring the cellular glutathione and malondialdehyde levels. Moreover, PA inhibited the TNF-α-induced phosphorylation of AKT, but not c-Jun N-terminal kinase, indicating that inhibition of survival signaling pathways activated by TNF-α may explain the effects of PA on TNF-α-induced cytotoxicity. The in vitro model established in this study may be used to screen drug candidates for suitability for the treatment of .Copyright © 2012 Elsevier Ltd. All rights reserved.

Keyword: NASH

Toll-like receptor 2 and cooperatively contribute to the development of nonalcoholic steatohepatitis through inflammasome activation in mice.

Innate immune signaling associated with Toll-like receptors (TLRs) is a key pathway involved in the progression of nonalcoholic steatohepatitis (). Here we show that both TLR2 and are required for activation of the inflammasome, interleukin (IL)-1α, and IL-1β, resulting in the progression of . Wild-type (WT) and TLR2(-/-) mice were fed a choline-deficient amino -defined (CDAA) diet for 22 weeks to induce . Bone marrow-transplanted TLR2 chimeric mice were generated after the recipient mice were lethally irradiated. Kupffer cells and hepatic stellate cells (HSCs) were isolated from WT mice and stimulated with TLR2 ligand and/or . WT mice on the CDAA diet developed profound steatohepatitis and liver fibrosis. In contrast, TLR2(-/-) mice had suppressed progression of . Although both Kupffer cells and HSCs respond to TLR2 ligand, TLR2 bone marrow chimeric mice demonstrated that Kupffer cells were relatively more important than HSCs in TLR2-mediated progression of . In vitro, alone did not increase TLR2 signaling-target genes, including cytokines and inflammasome components in Kupffer cells and HSCs. The TLR2 ligand increased Nod-like receptor protein 3, an inflammasome component, in Kupffer cells but not in HSCs. In the presence of TLR2 ligand, did induce caspase-1 activation and release of IL-1α and IL-1β in Kupffer cells; however, these effects were not observed in HSCs. In vivo, WT mice on the CDAA diet showed increased caspase-1 activation in the liver and elevated serum levels of IL-1α and IL-1β levels, which were suppressed in TLR2(-/-) mice.TLR2 and cooperatively activate the inflammasome in Kupffer cells and/or macrophages in the development of .Copyright © 2012 American Association for the Study of Liver Diseases.

Keyword: NASH

Reduction of oxidative stress attenuates lipoapoptosis exacerbated by hypoxia in human hepatocytes.

Chronic intermittent hypoxia, a characteristic of obstructive sleep apnea (OSA), is associated with the progression of simple hepatic steatosis to necroinflammatory hepatitis. We determined whether inhibition of a hypoxia-induced signaling pathway could attenuate hypoxia-exacerbated lipoapoptosis in human hepatocytes. The human hepatocellular carcinoma cell line (HepG2) was used in this study. (PA)-treated groups were used for two environmental conditions: Hypoxia (1% O2) and normoxia (20% O2). Following the treatment, the cell viability was determined by the 3,4-(5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium salt (MTS) assay, and the mechanism of lipoapoptosis was evaluated by Western blotting. Hypoxia exacerbated the suppression of hepatocyte growth induced by via activation of mitochondrial apoptotic pathways as a result of endoplasmic reticulum (ER) and oxidative stresses. Ammonium pyrrolidine dithiocarbamate, a scavenger of reactive oxygen species, attenuated the hypoxia-exacerbated lipoapoptosis in hepatocytes, whereas glycerol, which reduces ER stress, did not. This may have been because inhibition of oxidative stress decreases the expression of pro-apoptotic proteins, such as caspase 9 and cytochrome c. These results suggested that modulation of apoptotic signaling pathways activated by oxidative stress can aid in identifying novel therapeutic strategies for the treatment of nonalcoholic steatohepatitis () with OSA. Further in vivo studies are necessary to understand the pathophysiologic mechanism of with OSA and to prove the therapeutic effect of the modulation of the signaling pathways.

Keyword: NASH

Liraglutide ameliorates non-alcoholic steatohepatitis by inhibiting NLRP3 inflammasome and pyroptosis activation via mitophagy.

Non-alcoholic steatohepatitis () is a key step in the progression of non-alcoholic fatty liver disease (NAFLD), which causes serious health problems worldwide. The nucleotide-binding oligomerization domain, leucine-rich repeat-containing receptor-containing pyrin domain 3 (NLRP3) inflammasome and pyroptosis play crucial roles in the progression of . Our team has provided clinical evidence of the effects of glucagon-like peptide-1 (GLP-1) on the improvement in liver function and histological resolution of NAFLD. Preliminary work has demonstrated that GLP-1 inhibited NLRP3 inflammasome activation in a mouse model of NAFLD. We further explored the potential molecular mechanisms underlying the anti-inflammatory effect of liraglutide, a long-acting GLP-1 analog, in the treatment of . We established a HepG2 cell model of using double stimulation with and lipopolysaccharide to assess NLRP3 inflammasome and pyroptotic cell activity and to evaluate mitochondrial function and mitophagy. Liraglutide reduced lipid accumulation, inhibited NLRP3 inflammasome and pyroptosis activation, attenuated mitochondrial dysfunction and reactive oxygen species generation, augmented mitophagy in hepatocytes. Mitophagy inhibition with 3-methyladenine/PINK1-directed siRNA weakened the liraglutide-mediated suppression of inflammatory injury. We propose that liraglutide suppresses NLRP3 inflammasome-induced hepatocyte pyroptosis via mitophagy to slow the progression of .Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: NASH

Cangju Qinggan Jiangzhi Decoction Reduces the Development of NonAlcoholic Steatohepatitis and Activation of Kupffer Cells.

Nonalcoholic steatohepatitis () is defined as lipid accumulation with hepatic injury, inflammation and early to moderate fibrosis. Kupffer cells play a crucial role in promoting hepatic inflammation, which further facilitates the development of . Here we investigated the effects of Cangju Qinggan Jiangzhi decoction (CQJD) on high fat diet (HFD) and methionine-choline deficient (MCD) induced mouse pathogenesis.Mouse models were developed by HFD and MCD diet. The treated mice were divided into three groups: the control group (n = 10), the low-dose CQJD treatment group (n = 10) and the high-dose CQJD treatment group (n = 10). The hepatic injury, inflammation, and apoptotic molecules were evaluated by H&E staining, immunohistochemistry and real-time PCR. Kupffer cells were isolated from control mice and CQJD-treated mice after stimulation by lipopolysaccharide (LPS) and/or . The level of the inflammatory cytokines TNFα, IL1β, and CCL2 was measured by ELISA.The HFD-fed mice displayed significant metabolic, inflammatory, and oxidative stress-related alterations due to hepatic lipid accumulation. CQJD treatment largely normalized the hepatic injury, lowered the ALT/AST level, and reduced the severity of liver inflammation, as revealed by the decreased inflammatory cytokines levels. In vitro, CQJD blocked the activation of LPS- or -primed Kupffer cells in a dose-dependent manner. In the MCD diet-induced mice, similar therapeutic effects of CQJD were also observed.CQJD ameliorates mouse nonalcoholic steatohepatitis. The reduction in liver injury and inflammation induced by CQJD is associated with reduced activation of Kupffer cells. Our results suggest that CQJD is a promising therapeutic strategy in clinical steatohepatitis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: NASH

MSP is a negative regulator of inflammation and lipogenesis in ex vivo models of non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (), a metabolic disorder consisting of steatosis and inflammation, is considered the hepatic equivalent of metabolic syndrome and can result in irreversible liver damage. Macrophage-stimulating protein (MSP) is a hepatokine that potentially has a beneficial role in hepatic lipid and glucose metabolism via the activation of AMP-activated protein kinase (AMPK). In the current study, we investigated the regulatory role of MSP in the development of inflammation and lipid metabolism in various models, both in vitro and ex vivo. We observed that MSP treatment activated the AMPK signaling pathway and inhibited lipopolysaccharide (LPS)- and (PA)-induced gene expression of pro-inflammatory cytokines in primary mouse hepatocytes. In addition, MSP treatment resulted in a significant reduction in PA-induced lipid accumulation and inhibited the gene expression of key lipogenic enzymes in HepG2 cells. Upon short hairpin RNA-induced knockdown of RON (the membrane-bound receptor for MSP), the anti-inflammatory and anti-lipogenic effects of MSP were markedly ablated. Finally, to mimic ex vivo, we challenged bone marrow-derived macrophages with oxidized low-density lipoprotein (oxLDL) in combination with LPS. OxLDL+LPS exposure led to a marked inhibition of AMPK activity and a robust increase in inflammation. MSP treatment significantly reversed these effects by restoring AMPK activity and by suppressing pro-inflammatory cytokine gene expression and secretion under this condition. Taken together, these data suggest that MSP is an effective inhibitor of inflammation and lipid accumulation in the stressed liver, thereby indicating that MSP has a key regulatory role in .

Keyword: NASH

Hepatocellular carcinoma is accelerated by involving M2 macrophage polarization mediated by hif-1induced IL-10.

Obesity-related inflammation promotes cancer development. Tissue resident macrophages affect tumor progression and the tumor micro-environment favors polarization into alternatively activated macrophages (M2) that facilitate tumor invasiveness. Here, we dissected the role of western diet-induced in inducing macrophage polarization in a carcinogen initiated model of hepatocellular carcinoma (HCC). Adult C57BL/6 male mice received diethyl nitrosamine (DEN) followed by 24\xa0weeks of high fat-high cholesterol-high sugar diet (HF-HC-HSD). We assessed liver MRI and histology, serum ALT, AFP, liver triglycerides, and cytokines. Macrophage polarization was determined by IL-12/TNFα (M1) and CD163/CD206 (M2) expression using flow cytometry. Role of hif-1α-induced IL-10 was dissected in hepatocyte specific hif-1αKO and hif-1αdPA (over-expression) mice. The western diet-induced features of and accelerated HCC development after carcinogen exposure. Liver fibrosis and serum AFP were significantly increased in DEN + HF-HC-HSD mice compared to controls. Western diet resulted in macrophage (F4/80CD11b) infiltration to liver and DEN + HF-HC-HSD mice showed preferential increase in M2 macrophages. Isolated hepatocytes from western diet fed mice showed significant upregulation of the hypoxia-inducible transcription factor, hif-1α, and livers from hif-1α over-expressing mice had increased proportion of M2 macrophages. Primary hepatocytes from wild-type mice treated with DEN and showed activation of hif-1α and induction of IL-10, a M2 polarizing cytokine. IL-10 neutralization in hepatocyte-derived culture supernatant prevented M2 macrophage polarization and silencing hif-1α in macrophages blocked their M2 polarization. Therefore, our data demonstrate that accelerates HCC progression via upregulation of hif-1α mediated IL-10 polarizing M2 macrophages.

Keyword: NASH

Dual PPARα/γ agonist saroglitazar improves liver histopathology and biochemistry in experimental models.

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis () are common clinico-pathological conditions that affect millions of patients worldwide. In this study, the efficacy of saroglitazar, a novel PPARα/γ agonist, was assessed in models of NAFLD/.HepG2 cells treated with (PA;0.75 mM) showed decreased expression of various antioxidant biomarkers (SOD1, SOD2, glutathione peroxidase and catalase) and increased expression of inflammatory markers (TNFα, IL1β and IL6). These effects were blocked by saroglitazar, pioglitazone and fenofibrate (all tested at 10μM concentration). Furthermore, these agents reversed PA-mediated changes in mitochondrial dysfunction, ATP production, NFkB phosphorylation and stellate cell activation in HepG2 and HepG2-LX2 Coculture studies. In mice with choline-deficient high-fat diet-induced , saroglitazar reduced hepatic steatosis, inflammation, ballooning and prevented development of fibrosis. It also reduced serum alanine aminotransferase, aspartate aminotransferase and expression of inflammatory and fibrosis biomarkers. In this model, the reduction in the overall NAFLD activity score by saroglitazar (3\xa0mg/kg) was significantly more prominent than pioglitazone (25\xa0mg/kg) and fenofibrate (100\xa0mg/kg). Pioglitazone and fenofibrate did not show any improvement in steatosis, but partially improved inflammation and liver function. Antifibrotic effect of saroglitazar (4\xa0mg/kg) was also observed in carbon tetrachloride-induced fibrosis model.Saroglitazar, a dual PPARα/γ agonist with predominant PPARα activity, shows an overall improvement in . The effects of saroglitazar appear better than pure PPARα agonist, fenofibrate and PPARγ agonist pioglitazone.© 2017 Cadila Healthcare Ltd., Liver International Published by John Wiley & Sons Ltd.

Keyword: NASH

Development and validation of an in vitro 3D model of with severe fibrotic phenotype.

Nonalcoholic steatohepatitis represents a significant and rapidly growing unmet medical need. The development of novel therapies has been hindered in part, by the limitations of existing preclinical models. There is a strong need for physiologically relevant and liver fibrosis models that are characterized by better translational predictability. In this study, we used the InSphero 3D InSight three-dimensional (3D) human liver microtissue (3D-hLMT) system prepared by co-culturing primary human hepatocytes with hepatic stellate cells, Kupffer cells and endothelial cells to develop a model of with a severe fibrotic phenotype. In our model, (PA) induced a robust proinflammatory and profibrogenic phenotype in the 3D-hLMT. PA significantly increased several markers of the inflammatory and profibrotic process including gene expression of collagens, , tissue inhibitor of matrix metalloprotease 1 () and the stellate cell activation marker as well as secreted CXCL8 (IL8) levels. We also observed TGFβ pathway activation, increase in active collagen synthesis and significant overall increase in tissue damage in the 3D-hLMTs. Immunohistochemistry analysis demonstrated the upregulation of collagen, cleaved caspase 3 as well as of the PDGFRβ protein. We further validated the model using a phase 3 clinical compound, GS-4997, an apoptosis signal-regulating kinase 1 (ASK-1) inhibitor and showed that GS-4997 significantly decreased PA induced profibrotic and proinflammatory response in the 3D-hLMTs with decreases in apoptosis and stellate cell activation in the microtissues. Taken together we have established and validated an 3D-hLMT model with severe fibrotic phenotype that can be a powerful tool to investigate experimental compounds for the treatment of .

Keyword: NASH

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis () independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt . They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of .Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: NASH

Stimulated hepatic stellate cell promotes progression of hepatocellular carcinoma due to protein kinase R activation.

Hepatic stellate cells (HSCs) were reported to promote the progression of hepatocellular carcinoma (HCC), however its mechanism is uncertain. We previously reported that protein kinase R (PKR) in hepatocytes regulated HCC proliferation. In this study, we focused on the role of PKR in HSCs, and clarified the mechanism of its association with HCC progression. We confirmed the activation of PKR in a human HSC cell line (LX-2 cell). IL-1β is produced from HSCs stimulated by lipopolysaccharide (LPS) or which are likely activators of PKR in non-alcoholic steatohepatitis (). Production was assessed by real-time PCR and ELISA. C16 and small interfering RNA (siRNA) were used to inhibit PKR in HSCs. The HCC cell line (HepG2 cell) was cultured with HSC conditioning medium to assess HCC progression, which was evaluated by proliferation and scratch assays. Expression of PKR was increased and activated in stimulated HSCs, and IL-1β production was also increased molecular. Key molecules of the mitogen-activated protein kinase pathway were also upregulated and activated by LPS. Otherwise, PKR inhibition by C16 and PKR siRNA decreased IL-1β production. HCC progression was promoted by HSC-stimulated conditioning medium although it was reduced by the conditioning medium from PKR-inhibited HSCs. Moreover, also upregulated IL-1β expression in HSCs, and conditioning medium from -stimulated HSCs promoted HCC proliferation. Stimulated HSCs by activators of PKR in could play a role in promoting HCC progression through the production of IL-1β, via a mechanism that seems to be dependent on PKR activation.

Keyword: NASH

Increased expression of zinc finger protein 267 in non-alcoholic fatty liver disease.

Hepatocellular lipid accumulation is a hallmark of non-alcoholicfatty liver disease (NAFLD), which encompasses a spectrum ranging from simple steatosis to non-alcoholic steatohepatitis () and ultimately cirrhosis. Zinc finger protein 267 (ZNF267) belongs to the family of Kruppel-like transcription factors, which regulate diverse biological processes that include development, proliferation, and differentiation. We have previously demonstrated that ZNF267 expression is up-regulated in liver cirrhosis and is further increased in hepatocellular carcinoma (HCC). Here, we analyzed the expression of ZNF267 in tissue specimens of NAFLD patients and found a significant up-regulation compared to normal liver tissue. Noteworthy, ZNF267 mRNA was already significantly increased in steatotic liver tissue without inflammation. In line with this, incubation of primary human hepatocytes with induced a dose-dependent lipid accumulation and corresponding dose-dependent ZNF267 induction in vitro. Furthermore, hepatocellular lipid accumulation induced formation of reactive oxygen species (ROS), and also chemically induced ROS formation increased ZNF267 mRNA expression. In summary with previous findings, which revealed ZNF267 as pro-fibrogenic and pro-cancerogenic factor in chronic liver disease, the present study further suggests ZNF267 as promising therapeutic target particularly for NAFLD patients. In addition, it further indicates that hepatic steatosis per se has pathophysiological relevance and should not be considered as benign.

Keyword: NASH

The effect of enterolactone on liver lipid precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on lipid fractions fatty acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free fatty acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their fatty acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of fatty acids were evaluated by Western Blot.Enterolactone modified fatty acids composition in FFA, DAG and TAG fractions. In conjunction with lipid overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated fatty acids in each of the investigated lipid fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as .Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: NASH

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: NASH

Lipid-Induced Signaling Causes Release of Inflammatory Extracellular Vesicles From Hepatocytes.

Hepatocyte cellular dysfunction and death induced by lipids and macrophage-associated inflammation are characteristics of nonalcoholic steatohepatitis (). The fatty palmitate can activate death receptor 5 (DR5) on hepatocytes, leading to their death, but little is known about how this process contributes to macrophage-associated inflammation. We investigated whether lipid-induced DR5 signaling results in the release of extracellular vesicles (EVs) from hepatocytes, and whether these can induce an inflammatory macrophage phenotype.Primary mouse and human hepatocytes and Huh7 cells were incubated with palmitate, its metabolite lysophosphatidylcholine, or diluent (control). The released EV were isolated, characterized, quantified, and applied to macrophages. C57BL/6 mice were placed on chow or a diet high in fat, fructose, and cholesterol to induce . Some mice also were given the ROCK1 inhibitor fasudil; 2 weeks later, serum EVs were isolated and characterized by immunoblot and nanoparticle-tracking analyses. Livers were collected and analyzed by histology, immunohistochemistry, and quantitative polymerase chain reaction.Incubation of primary hepatocytes and Huh7 cells with palmitate or lysophosphatidylcholine increased their release of EVs, compared with control cells. This release was reduced by inactivating mediators of the DR5 signaling pathway or rho-associated, coiled-coil-containing protein kinase 1 (ROCK1) inhibition. Hepatocyte-derived EVs contained tumor necrosis factor-related apoptosis-inducing ligand and induced expression of interleukin 1β and interleukin 6 messenger RNAs in mouse bone marrow-derived macrophages. Activation of macrophages required DR5 and receptor-interacting protein kinase 1. Administration of the ROCK1 inhibitor fasudil to mice with reduced serum levels of EVs; this reduction was associated with decreased liver injury, inflammation, and fibrosis.Lipids, which stimulate DR5, induce release of hepatocyte EVs, which activate an inflammatory phenotype in macrophages. Strategies to inhibit ROCK1-dependent release of EVs by hepatocytes might be developed for the treatment of patients with .Copyright © 2016 AGA Institute. Published by Elsevier Inc. All rights reserved.

Keyword: NASH

Elafibranor restricts lipogenic and inflammatory responses in a human skin stem cell-derived model of .

Non-alcoholic steatohepatitis () is characterized by hepatocellular steatosis with concomitant hepatic inflammation. Despite its pandemic proportions, no anti- drugs have been approved yet. This is partially because drug development is decelerated due to the lack of adequate tools to assess the efficacy of potential new drug candidates. The present study describes the development and application of a new preclinical model for using hepatic cells generated from human skin-derived precursors. Exposure of these cells to lipogenic (insulin, glucose, fatty acids) and pro-inflammatory factors (IL-1β, TNF-α, TGF-β) resulted in a characteristic response, as indicated by intracellular lipid accumulation, modulation of -specific gene expression, increased caspase-3/7 activity and the expression and/or secretion of inflammatory markers, including CCL2, CCL5, CCL7, CCL8, CXCL5, CXCL8, IL1a, IL6 and IL11. The human relevance of the proposed model was verified by transcriptomics analyses that revealed commonly modulated genes and the identification of the same gene classes between the in vitro system and patients suffering from . The application potential of this in vitro model was demonstrated by testing elafibranor, a promising anti- compound currently under clinical phase III trial evaluation. Elafibranor attenuated in vitro key features of , and dramatically lowered lipid load as well as the expression and secretion of inflammatory chemokines, which in vivo are responsible for the recruitment of immune cells. This reduction in inflammatory response was NFκB-mediated. In summary, this human-relevant, in vitro system proved to be a sensitive testing tool for the investigation of novel anti- compounds.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: NASH

Sab (Sh3bp5) dependence of JNK mediated inhibition of mitochondrial respiration in induced hepatocyte lipotoxicity.

Sustained c-Jun N-terminal kinase (JNK) activation by saturated fatty acids plays a role in lipotoxicity and the pathogenesis of non-alcoholic steatohepatitis (). We have reported that the interaction of JNK with mitochondrial Sab leads to inhibition of respiration, increased reactive oxygen species (ROS), cell death and hepatotoxicity. We tested whether this pathway underlies (PA)-induced lipotoxicity in hepatocytes.Primary mouse hepatocytes (PMH) from adeno-shlacZ or adeno-shSab treated mice and HuH7 cells were used.In PMH, PA dose-dependently up to 1mM stimulated oxygen consumption rate (OCR) due to mitochondrial β-oxidation. At ⩾1.5mM, PA gradually reduced OCR, followed by cell death. Inhibition of JNK, caspases or treatment with antioxidant butylated hydroxyanisole (BHA) protected PMH against cell death. Sab knockdown or a membrane permeable Sab blocking peptide prevented PA-induced mitochondrial impairment, but inhibited only the late phase of both JNK activation (beyond 4h) and cell death. In PMH, PA increased p-PERK and its downstream target CHOP, but failed to activate the IRE-1α arm of the UPR. However, Sab silencing did not affect PA-induced PERK activation. Conversely, specific inhibition of PERK prevented JNK activation and cell death, indicating a major role upstream of JNK activation.The effect of p-JNK on mitochondria plays a key role in PA-mediated lipotoxicity. The interplay of p-JNK with mitochondrial Sab leads to impaired respiration, ROS production, sustained JNK activation, and apoptosis.Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: NASH

Loss of ULK1 increases RPS6KB1-NCOR1 repression of NR1H/LXR-mediated Scd1 transcription and augments lipotoxicity in hepatic cells.

Lipotoxicity caused by saturated fatty acids (SFAs) induces tissue damage and inflammation in metabolic disorders. SCD1 (stearoyl-coenzyme A desaturase 1) converts SFAs to mono-unsaturated fatty acids (MUFAs) that are incorporated into triglycerides and stored in lipid droplets. SCD1 thus helps protect hepatocytes from lipotoxicity and its reduced expression is associated with increased lipotoxic injury in cultured hepatic cells and mouse models. To further understand the role of SCD1 in lipotoxicity, we examined the regulation of Scd1 in hepatic cells treated with palmitate, and found that NR1H/LXR (nuclear receptor subfamily 1 group H) ligand, GW3965, induced Scd1 expression and lipid droplet formation to improve cell survival. Surprisingly, ULK1/ATG1 (unc-51 like kinase) played a critical role in protecting hepatic cells from SFA-induced lipotoxicity via a novel mechanism that did not involve macroautophagy/autophagy. Specific loss of Ulk1 blocked the induction of Scd1 gene transcription by GW3965, decreased lipid droplet formation, and increased apoptosis in hepatic cells exposed to palmitate. Knockdown of ULK1 increased RPS6KB1 (ribosomal protein S6 kinase, polypeptide 1) signaling that, in turn, induced NCOR1 (nuclear receptor co-repressor 1) nuclear uptake, interaction with NR1H/LXR, and recruitment to the Scd1 promoter. These events abrogated the stimulation of Scd1 gene expression by GW3965, and increased lipotoxicity in hepatic cells. In summary, we have identified a novel autophagy-independent role of ULK1 that regulates NR1H/LXR signaling, Scd1 expression, and intracellular lipid homeostasis in hepatic cells exposed to a lipotoxic environment.

Keyword: NASH

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis () induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver-body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of , both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: NASH

Fluvastatin attenuates hepatic steatosis-induced fibrogenesis in rats through inhibiting paracrine effect of hepatocyte on hepatic stellate cells.

Non-alcoholic steatohepatitis () is associated with hepatic fibrogenesis. Despite well-known cholesterol-lowering action of statins, their mechanisms against -mediated fibrogenesis remain unclear. This study aimed at investigating the in vitro and in vivo anti-fibrotic properties of fluvastatin (Flu).Palmitate (PA)-induced changes in intracellular hydrogen peroxide levels in primary rat hepatocytes (PRHs) and human hepatoma cell line (HepG2) were quantified by dichlorofluorescein diacetate (DCF-DA) dye assay, whereas changes in expressions of NADPH oxidase gp91 (phox) subunit, α-smooth muscle actin (α-SMA), and NFκB p65 nuclear translocation were quantified with Western blotting. Quantitative real-time polymerase chain reaction (q-PCR) was used to investigate mRNA expressions of pro-inflammatory genes (ICAM-1, IL-6, TNF-α). Conditioned medium (CM) from PA-treated PRHs was applied to cultured rat hepatic stellate cell line, HSC-T6, with or without Flu-pretreatment for 2 h. Pro-fibrogenic gene expressions (COL1, TIMP-1, TGF-β1, α-SMA) and protein expression of α-SMA were analyzed. In vivo study using choline-deficient L-amino defined (CDAA) diet-induced rat model was performed by randomly assigning Wistar rats (n = 28) to normal controls (n = 4), CDAA diet with vehicles, and CDAA diet with Flu (5 mg/kg or 10 mg/kg) (n = 8 each) through gavage for 4 or 8 weeks. Livers were harvested for histological, Western blot (α-SMA), and q-PCR analyses for expressions of pro-inflammatory (IL-6, iNOS, ICAM-1) and pro-fibrogenic (Col1, α-SMA, TIMP-1) genes.In vitro, Flu (1-20 μM) inhibited PA-induced free-radical production, gp91 (phox) expression, and NFκB p65 translocation in HepG2 and PRHs, while CM-induced α-SMA protein expression and pro-fibrogenic gene expressions in HSC-T6 were suppressed in Flu-pretreated cells compared to those without pretreatment. Moreover, α-SMA protein expression was significantly decreased in HSC-T6 cultured with CM from PA-Flu-treated PRHs compared to those cultured with CM from PA-treated PRHs. Flu also reduced steatosis and fibrosis scores, α-SMA protein expression, mRNA expression of pro-inflammatory and pro-fibrogenic genes in livers of CDAA rats.We demonstrated PA-induced HSC activation through paracrine effect of hepatocyte in vitro that was significantly suppressed by pre-treating HSC with Flu. In vivo, Flu alleviated steatosis-induced HSC activation and hepatic fibrogenesis through mitigating inflammation and oxidative stress, suggesting possible therapeutic role of Flu against .

Keyword: NASH

Activation of the GP130-STAT3 axis and its potential implications in nonalcoholic fatty liver disease.

The status of the GP130-STAT3 signaling pathway in humans with nonalcoholic fatty liver disease (NAFLD) and its relevance to disease pathogenesis are unknown. The expression of the gp130-STAT3 axis and gp130 cytokine receptors were studied in subjects with varying phenotypes of NAFLD including nonalcoholic steatohepatitis () and compared with lean and weight-matched controls without NAFLD. Gp130 and its downstream signaling element (Tyk2 and STAT3) expression were inhibited in obese controls whereas they were increased in NAFLD. IL-6 levels were increased in and correlated with gp130 expression (P < 0.01). Palmitate inhibited gp130-STAT3 expression and signaling. IL-6 and palmitate inhibited hepatic insulin signaling via STAT3-dependent and independent mechanisms, respectively. STAT3 overexpression reversed palmitate-induced lipotoxicity by increasing autophagy (ATG7) and decreasing endoplasmic reticulum stress. These data demonstrate that the STAT3 pathway is activated in NAFLD and can worsen insulin resistance while protecting against other lipotoxic mechanisms of disease pathogenesis.Copyright © 2015 the American Physiological Society.

Keyword: NASH

Thioacetamide potentiates high cholesterol and high fat diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing .

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis () and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and liver anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: NASH

[Effect of free fatty on NALP3 inflammasome signaling pathway in THP-1 macrophages].

To investigate the potential effect of NALP3 inflammasome on the occurrence and development of nonalcoholic steatohepatitis ().THP-1 macrophages were cultured for 24 h by at various concentrations. The THP-1 macrophages were pretreated with N-acetyl-cysteine at different doses for 24 h before the cultivation. ROS production was determined by flow cytometry. The expression of IL- 1β was detected by ELISA; the expressions of NALP3 protein and caspase-1 protein were detected by immunofluorescence; NALP3, ASC, and caspase-1 mRNA were measured by real-time PCR.Compared with the THP-1 macrophages without , the level of ROS, NALP3 protein and caspase-1 protein, and the expression of IL-1β were increased after treatment in a dose dependent manner (P<0.05). Compared with the THP-1 macrophages with (400 μmol/L), the level of NALP3 mRNA (P<0.05), the level of NALP3 protein and caspase-1 protein (P<0.05), the expression of IL-1β (P<0.05) were decreased after preadministration of N-acetyl-cysteine in a dose dependent manner.ROS induced by free fatty can regulate the activation of NALP3 inflammasome signaling pathway leading to the release of inflammatory cytokines. This pathway may be the possible mechanism of .

Keyword: NASH

Augmented hepatic Toll-like receptors by fatty acids trigger the pro-inflammatory state of non-alcoholic fatty liver disease in mice.

There is considerable evidence that intestinal microbiota are involved in the development of metabolic syndromes and, consequently, with the development of non-alcoholic fatty liver disease (NAFLD). Toll-like receptors (TLRs) are essential for the recognition of microbiota. However, the induction mechanism of TLR signals through the gut-liver axis for triggering the development of non-alcoholic steatohepatitis () or NAFLD remains unclear. In this study, we investigated the role of (PA) in triggering the development of a pro-inflammatory state of NAFLD.Non-alcoholic fatty liver disease was induced in mice fed a high fat diet (HFD). The mice were killed and the expression of TLRs, tumor necrosis factor (TNF), interleukin (IL)-1β, and phospho-interleukin-1 receptor-associated kinase 1 in the liver and small intestine were assessed. In addition, primary hepatocytes and Kupffer cells were treated with PA, and the direct effects of PA on TLRs induction by these cells were evaluated.The expression of inflammatory cytokines such as TNF, IL-1β, and TLR-2, -4, -5, and -9 was increased in the liver, but decreased in the small intestine of HFD-fed mice in vivo. In addition, the expression of TLRs in primary hepatocytes and Kupffer cells was increased by treatment with PA.In the development of the pro-inflammatory state of NAFLD, PA triggers the expression of TLRs, which contribute to the induction of inflammatory cytokines through TLR signals by intestinal microbiota.© 2013 The Japan Society of Hepatology.

Keyword: NASH

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis () and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the liver of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in liver cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the liver steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit by its action on both inflammatory response and liver lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: NASH

Oxysterols induce mitochondrial impairment and hepatocellular toxicity in non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a chronic hepatic disorder affecting up to 25% of the general population. Several intracellular events leading to NAFLD and progression to non-alcoholic steatohepatitis () have been identified, including lipid accumulation, mitochondrial dysfunction and oxidative stress. Emerging evidence links both hepatic free fatty acids (FFAs) and cholesterol (FC) accumulation in NAFLD development; in particular oxysterols, the oxidative products of cholesterol, may contribute to liver injury. We performed a targeted lipidomic analysis of oxysterols in the liver of male Wistar rats fed a high-fat (HF), high-cholesterol (HC) or high-fat/high-cholesterol (HF/HC) diet. Both HF and HC diets caused liver steatosis, but the HF/HC diet resulted in steatohepatitis with associated mitochondrial dysfunction. Above all, the oxysterol cholestane-3beta,5alpha,6beta-triol (triol) was particularly increased in the liver of rats fed diets rich in cholesterol. To verify the molecular mechanism involved in mitochondrial dysfunction and hepatocellular toxicity, Huh7 and primary rat hepatocytes were exposed to (PA) and/or oleic (OA), with or without triol. This compound induced apoptosis in cells co-exposed to both PA and OA, and this was associated with impaired mitochondrial respiration as well as down-regulation of PGC1-alpha, mTFA and NRF1.In conclusion, our data show that hepatic free fatty or oxysterols accumulation per se induce low hepatocellular toxicity. On the contrary, hepatic accumulation of both fatty acids and toxic oxysterols such as triol are determinant in the impairment of mitochondrial function and biogenesis, contributing to liver pathology in NAFLD.Copyright © 2014. Published by Elsevier Inc.

Keyword: NASH

induces production of proinflammatory cytokine interleukin-8 from hepatocytes.

Obesity and the metabolic syndrome are closely correlated with hepatic steatosis. Simple hepatic steatosis in nonalcoholic fatty liver disease can progress to nonalcoholic steatohepatitis (), which can be a precursor to more serious liver diseases, such as cirrhosis and hepatocellular carcinoma. The pathogenic mechanisms underlying progression of steatosis to remain unclear; however, inflammation, proinflammatory cytokines, and oxidative stress have been postulated to play key roles. We previously reported that patients with have elevated serum levels of proinflammatory cytokines, such as interleukin-8 (IL-8), which are likely to contribute to hepatic injury. This study specifically examines the effect of hepatic steatosis on IL-8 production. We induced lipid accumulation in hepatocytes (HepG2, rat primary hepatocytes, and human primary hepatocytes) by exposing them to pathophysiologically relevant concentrations of to simulate the excessive influx of fatty acids into hepatocytes. Significant fat accumulation was documented morphologically by Oil Red O staining in cells exposed to , and it was accompanied by an increase in intracellular triglyceride levels. Importantly, was found to induce significantly elevated levels of biologically active neutrophil chemoattractant, IL-8, from steatotic hepatocytes. Incubation of the cells with palmitate led to increased IL-8 gene expression and secretion (both mRNA and protein) through mechanisms involving activation of nuclear factor kappaB (NF-kappaB) and c-Jun N-terminal kinase/activator protein-1.These data demonstrate for the first time that lipid accumulation in hepatocytes can stimulate IL-8 production, thereby potentially contributing to hepatic inflammation and consequent liver injury.

Keyword: NASH

Upregulated absorption of dietary acids with changes in intestinal transporters in non-alcoholic steatohepatitis ().

is an important risk factor for the pathogenesis of non-alcoholic steatohepatitis (), but changes in intestinal absorption in are unclear. The aim of this study was to clarify changes in intestinal absorption and their association with the pathogenesis of .A total of 106 participants were recruited to the study, of whom 33 were control subjects (control group), 32 were patients with Brunt stage 1-2 [early (e-)], and 41 were patients with Brunt stage 3-4 [advanced (a-)]. C-labeled palmitate was administered directly into the duodenum of all participants by gastrointestinal endoscopy. Breath CO levels were measured to quantify absorption, and serum Apolipoprotein B-48 (ApoB-48) concentrations were measured after a test meal to quantify absorbed chylomicrons. Expressions of fatty (FA) transporters were also examined. The associations of breath CO levels with hepatic steatosis, fibrosis and insulin resistance was evaluated using laboratory data, elastography results and liver histology findings.Overall, CO excretion was significantly higher in e- patients than in the control subjects and a- patients (P\xa0<\xa00.01). e- patients had higher serum ApoB-48 levels, indicating increased transport via chylomicrons in these patients. Jejunal mRNA and protein expressions of microsomal triglyceride transfer protein and cluster of differentiation 36 were also increased in both patient groups. The CO excretion of e- patients was significantly correlated with the degree of hepatic steatosis, fibrosis and insulin resistance (P\xa0=\xa00.005, P\xa0<\xa00.001, P\xa0=\xa00.019, respectively).Significantly upregulated absorption by activation of its transporters was evident in patients with , and clinical progression of was related to absorption. These dietary changes are associated with the onset and progression of .

Keyword: NASH

Hepatitis C Virus Infection Increases c-Jun N-Terminal Kinase (JNK) Phosphorylation and Accentuates Hepatocyte Lipoapoptosis.

BACKGROUND Hepatitis C virus (HCV) infection and metabolic diseases including nonalcoholic steatohepatitis () exhibit a complex interplay. Although free fatty -mediated apoptosis is a prominent feature of , the impact of HCV infection on hepatocyte lipotoxicity has remained largely unexplored. The study aimed at identifying whether infection by HCV affected the apoptotic pathway in hepatocytes during fatty assault. MATERIAL AND METHODS OR6 cells, which are derived from human hepatocellular carcinoma Huh-7 cells and harbor a full-length HCV RNA genome replication system, were treated with palmitate. Apoptosis was examined by 4\',6-diamidino-2-phenylindole staining. Activation and expression of JNK, Bim, cIAP-1, and Mcl-1 were examined by immunoblotting. mRNA expression of CHOP, a major player in endoplasmic reticulum stress-mediated apoptosis, was assessed by real-time PCR. RESULTS Palmitate-induced hepatocyte apoptosis was significantly enhanced in OR6 cells compared to cured cells, in which the HCV genome had been eradicated by treatment with interferon-α. Although basal expression of CHOP mRNA was enhanced in OR6 cells compared to cured cells, it was similarly upregulated in both cell lines following palmitate treatment. Notably, palmitate-induced JNK phosphorylation was accentuated in OR6 cells compared to cured cells. Inhibition of JNK with SP600125 attenuated palmitate-induced apoptosis. Palmitate-mediated upregulation of BH3-only protein Bim, which acts downstream of JNK, was also enhanced in OR6 cells compared to cured cells. In contrast, Mcl-1 and cIAP-1 were equally reduced in OR6 cells and cured cells following palmitate treatment. CONCLUSIONS These findings suggest that during lipoapoptosis, HCV infection may enhance hepatocyte toxicity by increasing JNK phosphorylation.

Keyword: NASH

Ferroptosis affects the progression of non-alcoholic steatohepatitis via the modulation of lipid peroxidation-mediated cell death in mice.

Oxidative stress and its-associated lipid peroxidation play a key role in non-alcoholic steatohepatitis (). Ferroptosis is a recently recognized type of cell death characterized by an iron dependent and lipid peroxidation-mediated non-apoptotic cell death. We demonstrate the impact of ferroptosis on the progression of induced by methionine/choline-deficient diet (MCD) feeding for 10 days. RSL-3 (a ferroptosis inducer) treatment showed decreased hepatic expression of GPX4, and conversely increased 12/15-lipoxygenase, and apoptosis inducing factor, indicating that ferroptosis plays a key role in -related lipid peroxidation and its-associated cell death. Consistently, levels of serum biochemical, hepatic steatosis, inflammation, and apoptosis in MCD-fed mice were exacerbated with RSL-3 treatment. However, MCD-fed mice treated with sodium selenite (a GPX4 activator) showed increase of hepatic GPX4, accompanied by reduced severity. To chelate iron, deferoxamine mesylate salt was used. Administration of deferoxamine mesylate salt significantly reduced severity and abolished the harmful effects of RSL-3 in MCD-fed mice. Finally, treatment with Liproxstatin-1 (a ferroptosis inhibitor) repressed hepatic lipid peroxidation, and its associated cell death, resulting in decreased severity. Consistent with the in vivo findings, modulation of ferroptosis /GPX4 affected hepatocellular death in -induced in vitro milieu. We conclude that GPX4 and its-related ferroptosis might play a major role in the development of .Copyright © 2019. Published by Elsevier Inc.

Keyword: NASH

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/.

Keyword: NASH

Bee\'s honey attenuates non-alcoholic steatohepatitis-induced hepatic injury through the regulation of thioredoxin-interacting protein-NLRP3 inflammasome pathway.

We aim to examine whether honey ameliorates hepatic injury in non-alcoholic steatohepatitis () animal and cell line steatosis models. was induced in female Sprague-Dawley rat by 8-week feeding with a high-fat diet. During the experiment, 5\xa0g/kg honey was intragastrically fed daily. Rat normal hepatocyte BRL-3A cell was treated with sodium palmitate (SP) to induce steatosis in the absence or presence of honey pre-treatment or specific siRNA/overexpress plasmid of thioredoxin-interacting protein (TXNIP) or antagonist/agonist of Nod-like receptor protein 3 (NLRP3).Honey significantly improved the high-fat-diet-induced hepatic injury, steatosis, fibrosis, oxidative stress, and inflammation in rats. Honey also inhibited the overexpression of TXNIP and the activation of NLRP3 inflammasome. These effects were replicated in BRL-3A cell line which showed that the down-regulation of TXNIP or inhibition of NLRP3 contributed to the suppression of NLRP3 inflammasome activation, inflammation, and re-balanced lipid metabolism. In contrast, overexpression of TXNIP or agonism of NLRP3 exacerbated the cellular damage induced by SP.Suppression of the TXNIP-NLRP3 inflammasome pathway may partly contribute to the amelioration of hepatic injury during the progression of by honey. Targeting hepatic TXNIP-NLRP3 inflammasome pathway is a potential therapeutic way for the prevention and treatment of .

Keyword: NASH

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis () is characterized by lipid accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce and liver fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in inflammation and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated lipid accumulation, inflammation and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents and fibrosis via inhibition of oxidative stress and inflammation, highlighting polydatin as a potential therapeutic agent for prevention and treatment of .

Keyword: NASH

elicits hepatic stellate cell activation through inflammasomes and hedgehog signaling.

Activation of hepatic stellate cells (HSCs) plays a pivotal role at the center of the fibrogenic progression in nonalcoholic steatohepatitis (). However, it is poorly understood that how various molecules interact within HSCs during the progression of to fibrosis. The aim of the present study is to delineate how inflammasome molecules, hedgehog signaling and autophagy provoke HSC activation using (PA) as a major insult.Inflammasome activation, hedgehog signaling activity and autophagy in PA-exposed HSCs were determined to investigate their role in activation of human and rodent HSC lines or primary HSCs.PA treatment elicited HSC activation reflected by increased mRNA levels of transforming growth factor-β1, connective tissue growth factor, tissue inhibitor of metalloproteinase-1 and procollagen type I (α1). In addition, expression levels of NOD-like receptor protein 3 (NLRP3) and hedgehog signaling transcription factor Gli-1 were increased in PA-exposed HSCs. It\'s evident that PA treatment resulted in increased production of light chain 3-II and autophagosomes, as well as enhanced autophagy flux reflected by transduction of an adeno-associated viral vector. Whereas, reduced autophagy, which is often seen in the late stage of , provoked inflammasome activation. Moreover, suppressing the Hh signaling pathway by LDE225 blocked production of light chain 3-II and autophagy flux.Saturated fatty acids, such as PA, stimulate HSC activation through inflammasomes and hedgehog signaling. Meanwhile, compromised autophagy may facilitate HSC activation, implicating valuable candidates for pharmacologic intervention against the progression of fibrogenesis in .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: NASH

Adenosine A2a receptor stimulation blocks development of nonalcoholic steatohepatitis in mice by multilevel inhibition of signals that cause immunolipotoxicity.

Lipotoxicity and immunoinflammation are associated with the evolution of steatosis toward nonalcoholic steatohepatitis (). This study reports the ability of adenosine A2a receptor (A2aR) activation to inhibit development by modulating the responses of CD4 T-helper (Th) cells to avoid an immuno-mediated potentiation of lipotoxicity. The effect of the A2aR agonist CGS21680 on immunoinflammatory signals, CD4Th cell infiltration and immunolipotoxicity was analyzed in steatotic C57BL/6 mice fed with a methionine-choline-deficient (MCD) diet and in mouse hepatocytes exposed to (PA). CGS21680 inhibited development in steatotic mice and decreased cytokines and chemokines involved in Th cell recruitment or polarization (namely CXCL10, CCL2, tumor necrosis factor alfa [TNFα], tumor growth factor [TGFβ], and IL-12). CGS21680 also reduced the expansion of Th17, Th22, and Th1 cells and increased the immunosuppressive activity of T regulatory cells. In PA-treated mice hepatocytes, CGS21680 inhibited the production of CXCL10, TNFα, TGFβ, IL-12, and CCL2; CGS21680 also prevented JNK-dependent lipotoxicity and its intensification by IL-17 or IL-17 plus IL-22 through Akt/PI3-kinase stimulation and inhibition of the negative regulator of PI3-kinase, (phosphatase and tensin homologue deleted from chromosome 10 (PTEN), which is upregulated by IL-17. In MCD livers, CGS21680 reduced JNK activation and PTEN expression and increased Akt phosphorylation. In conclusion, A2aR stimulation inhibited development by reducing Th17\xa0cell expansion and inhibiting the exacerbation of the IL-17-induced JNK-dependent lipotoxicity. These data promote the implementation of further studies to evaluate the potential clinical application of A2aR agonists that, by being able to function as both cytoprotective and immunomodulatory agents, could efficiently antagonize\xa0the multi-faced pathogenesis of .Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: NASH

Oleic protects saturated fatty mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis.

Aim This study aims to demonstrate the protective effects of monounsaturated oleic (OA) against saturated (PA) induced cellular lipotoxicity in hepatocytes and rats with non-alcoholic steatohepatitis ().Human hepatoma cell line HepG2 cells and neonatal rat primary hepatocytes were treated with PA or/and OA for 24\u202fh. SD rats were fed with high fat diet (HFD) to induce . From the 16th w, the HFD was full or half replaced by olive oil to observe the protective effects.In vitro, OA substantially alleviated PA induced cellular apoptosis, oxidative stress, ER stress, mitochondrial dysfunction, as well as inflammation in hepatocytes. In vivo, only olive oil supplementation had no detrimental effects, while HFD developed in normal rats. Full replacement of HFD with olive oil had profoundly reversed . Noteworthily, half replacement of HFD with olive oil (a mixed diet) has ameliorated injury as well. It strikingly changed the hepatic histology from macrovesicular-steatosis into entire microvesicular-steatosis, and significantly reduced inflammation, ballooning and fibrosis.Our study has demonstrated in both hepatocytes and rats that oleic acids had great potential to combat the saturated fatty acids induced hepatic lipotoxicity. Only half replacement of HFD by monounsaturated fatty acids rich diet still had significant therapeutic outcome in rats. Redirecting the toxic saturated fatty acids into triglyceride storage and reduction of cholesterol accumulation might be the possible explanation of OA driven protection in this scenario.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: NASH

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis () is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of . Although silibinin is used for the treatment of in clinical, its effect on CFLAR-JNK pathway in remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: NASH

Characteristics of hepatic fatty compositions in patients with nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is closely related to insulin resistance and lipid metabolism. Recent studies have suggested that the quality of fat accumulated in the liver is associated with the development of nonalcoholic steatohepatitis (). In this study, we investigated the fatty composition in liver tissue and its association with the pathology in NAFLD patients.One hundred and three patients diagnosed with NAFLD [simple steatosis (SS): 63, : 40] were examined and their hepatic fatty acids were measured using gas chromatography. In addition, relationships between the composition and composition ratios of various fatty acids and patient backgrounds, laboratory test values, histology of the liver, and expression of fat metabolism-related enzymes were investigated.The C16:1n7 content, the C16:1n7/C16:0 and C18:1n9/C18:0 ratios were increased and the C18:0/C16:0 ratio was decreased in the group. The C18:0/C16:0 and C18:1n9/C18:0 ratios were associated with the steatosis score in liver tissue, and the C16:1n7/C16:0 ratio was associated with the lobular inflammation score. The expressions levels of genes: SCD1, ELOVL6, SREBP1c, FAS and PPARγ were enhanced in the group. In multivariate analysis, the C18:0/C16:0 ratio was the most important factor that was correlated with the steatosis score. In contrast, the C16:1n7/C16:0 ratio was correlated with lobular inflammation.The fatty composition in liver tissue and expression of genes related to fatty metabolism were different between the SS and groups, suggesting that the acceleration of fatty metabolism is deeply involved in pathogenesis of .© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: NASH

Fatty and endotoxin activate inflammasomes in mouse hepatocytes that release danger signals to stimulate immune cells.

The pathogenesis of nonalcoholic steatohepatitis () and inflammasome activation involves sequential hits. The inflammasome, which cleaves pro-interleukin-1β (pro-IL-1β) into secreted IL-1β, is induced by endogenous and exogenous danger signals. Lipopolysaccharide (LPS), a toll-like receptor 4 ligand, plays a role in and also activates the inflammasome. In this study, we hypothesized that the inflammasome is activated in by multiple hits involving endogenous and exogenous danger signals. Using mouse models of methionine choline-deficient (MCD) diet-induced and high-fat diet-induced , we found up-regulation of the inflammasome [including NACHT, LRR, and PYD domains-containing protein 3 (NALP3; cryopyrin), apoptosis-associated speck-like CARD-domain containing protein, pannexin-1, and pro-caspase-1] at the messenger RNA (mRNA) level increased caspase-1 activity, and mature IL-1β protein levels in mice with steatohepatitis in comparison with control livers. There was no inflammasome activation in mice with only steatosis. The MCD diet sensitized mice to LPS-induced increases in NALP3, pannexin-1, IL-1β mRNA, and mature IL-1β protein levels in the liver. We demonstrate for the first time that inflammasome activation occurs in isolated hepatocytes in steatohepatitis. Our novel data show that the saturated fatty (FA) (PA) activates the inflammasome and induces sensitization to LPS-induced IL-1β release in hepatocytes. Furthermore, PA triggers the release of danger signals from hepatocytes in a caspase-dependent manner. These hepatocyte-derived danger signals, in turn, activate inflammasome, IL-1β, and tumor necrosis factor α release in liver mononuclear cells.Our novel findings indicate that saturated FAs represent an endogenous danger in the form of a first hit, up-regulate the inflammasome in , and induce sensitization to a second hit with LPS for IL-β release in hepatocytes. Furthermore, hepatocytes exposed to saturated FAs release danger signals that trigger inflammasome activation in immune cells. Thus, hepatocytes play a key role in orchestrating tissue responses to danger signals in .Copyright © 2011 American Association for the Study of Liver Diseases.

Keyword: NASH

Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic fatty liver to nonalcoholic steatohepatitis.

Activation of inflammation is an important mechanism in the development of nonalcoholic steatohepatitis (). This study aims to delineate how mitophagy affects NLRP3 inflammasome activation in hepatic lipotoxicity. Mice were fed a high fat/calorie diet (HFCD) for 24 weeks. Primary rat hepatocytes were treated with (PA) for various periods of time. Mitophagy was measured by protein levels of LC3II and P62. NLRP3, caspase-1, interleukin (IL)-18, and IL-1β at mRNA and protein levels were used as indicators of inflammasome activation. Along with steatotic progression in HFCD-fed mice, ratio of LC3II/β-actin was decreased concurrently with increased levels of liver P62, NLRP3, caspase-1, IL-1β, IL-18, and serum IL-1β levels in late-stage . PA treatment resulted in mitochondrial oxidative stress and initiated mitophagy in primary hepatocytes. The addition of cyclosporine A did not change LC3II/Τοmm20 ratios; but P62 levels were increased after an extended duration of PA exposure, indicating a defect in autophagic activity. Along with impaired mitophagy, mRNA and protein levels of NLRP3, caspase-1, IL-18 and IL-1β were upregulated by PA treatment. Pretreatment with MCC950, N-acetyl cysteine or acetyl-L-carnitine reversed inflammasome activation and a pyroptotic cascade. Additionally, mitophagic flux was partially recovered as indicated by increases in LC3II/Tomm20 ratio, parkin, and PINK1 expression, and decreased P62 expression. The findings suggest that impaired mitophagy triggers hepatic NLRP3 inflammasome activation in a murine model and primary hepatocytes. The new insights into inflammasome activation through mitophagy advance our understanding of how fatty acids elicit lipotoxicity through oxidant stress and autophagy in mitochondria.

Keyword: NASH

Chronic administration of saturated fats and fructose differently affect SREBP activity resulting in different modulation of Nrf2 and Nlrp3 inflammasome pathways in mice liver.

The overconsumption of both saturated fats and fructose in the modern society has been related to the development of nonalcoholic fatty liver disease (NAFLD). However, the specific contribution of individual dietary components on the progression of NAFLD to nonalcoholic steatohepatitis () has been poorly investigated. Therefore, the aim of our study was to investigate the dissimilar effects of these two dietary components on selected proinflammatory and antioxidant pathways in the liver of C57BL/6 mice fed a standard (SD), a 45% saturated fat (HFAT) or a 60% fructose (HFRT) diet for 12 weeks. HFAT diet evoked systemic metabolic alterations and overweight, not observed in HFRT mice. However, HFRT mice had a greater hepatic triglyceride deposition with increased ratio of triacylglycerols containing the compared to HFAT, as assessed by liquid chromatography-mass spectrometry analysis. This effect is due to the higher activation of the SCAP/SREBP1c lipogenic pathway by HFRT feeding. In addition, we found inhibition of Keap1/Nrf2 antioxidant signaling and more robust stimulation of the Nlrp3 inflammasome pathway in the livers of HFRT-fed mice when compared with HFAT-fed mice, which is consistent with the recent finding that palmitate and SREBP1c are implicated in hepatic oxidative stress and inflammation. These effects were associated with increased hepatic inflammation, as confirmed by high expression of markers of leukocyte infiltration in the HFRT group. Thus, we hypothesize an amplifying loop among lipogenesis, palmitate, Nrf2 and Nlrp3 that leads to a higher risk of NAFLD progression to in a high-fructose diet compared to a high-saturated fat intake.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: NASH

Inhibition of NLRP3 inflammasome by thioredoxin-interacting protein in mouse Kupffer cells as a regulatory mechanism for non-alcoholic fatty liver disease development.

NOD-like receptor (NLR) NLRP3 inflammasome activation has been implicated in the progression of non-alcoholic fatty liver disease (NAFLD) from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (). It has been also shown that (PA) activates NLRP3 inflammasome and promotes interleukin-1β (IL-1β) secretion in Kupffer cells (KCs). However, the specific mechanism of the NLRP3 inflammasome activation is unclear. We studies the molecular mechanisms by investigating the roles of Thioredoxin-interacting protein (TXNIP) and NLRP3 on NAFLD development in patients, high-fat diet (HFD)-induced NAFL and methionine choline deficient (MCD) diet-induced in wild type (WT), TXNIP-/- (thioredoxin-interacting protein) and NLRP3-/- mice, and isolated KCs. We found that the expressions of NLRP3 and TXNIP in human liver tissues were higher in group than in NAFL group. Furthermore, co-immunoprecipitation analyses show that activation of the TXNIP-NLRP3 inflammasome protein complex occurred in KCs of WT mice rather than NAFL WT mice, thus suggesting that the formation and activation of this protein complex is mainly involved in the development of . NLRP3-/- mice exhibited less severe than WT mice in MCD diet model, whereas TXNIP deficiency enhanced NLRP3 inflammasome activation and exacerbated liver injury. PA triggered the activation and co-localization of the NLRP3 inflammasome protein complex in KCs isolated from WT and TXNIP-/- but not NLRP3-/- mice, and most of the complex co-localized with mitochondria of KCs following PA stimulation. Taken together, our novel findings indicate that TXNIP plays a protective and anti-inflammatory role in the development of NAFLD through binding and suppressing NLRP3.

Keyword: NASH

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the liver of high-fat and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in liver sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of .

Keyword: NASH

Uncoupling protein 2 regulates -induced hepatoma cell autophagy.

Mitochondrial uncoupling protein 2 (UCP2) is suggested to have a role in the development of nonalcoholic steatohepatitis (). However, the mechanism remains unclear. Autophagy is an important mediator of many pathological responses. This study aims to investigate the relationship between UCP2 and hepatoma cells autophagy in - (PA-) induced lipotoxicity. H4IIE cells were treated with (PA), and cell autophagy and apoptosis were examined. UCP2 expression, in association with LC3-II and caspase-3, which are indicators of cell autophagy and apoptosis, respectively,was measured. Results demonstrated that UCP2 was associated with autophagy during PA-induced hepatic carcinoma cells injury. Tests on reactive oxygen species (ROS) showed that UCP2 overexpression strongly decreases PA-induced ROS production and apoptosis. Conversely, UCP2 inhibition by genipin or UCP2 mRNA silencing enhances PA-induced ROS production and apoptosis. Autophagy partially participates in this progress. Moreover, UCP2 was associated with ATP synthesis during PA-induced autophagy. In conclusion, increasing UCP2 expression in hepatoma cells may contribute to cell autophagy and antiapoptotic as result of fatty injury. Our results may bring new insights for potential therapies.

Keyword: NASH

Hepatocyte X-box binding protein 1 deficiency increases liver injury in mice fed a high-fat/sugar diet.

Fatty liver is associated with endoplasmic reticulum stress and activation of the hepatic unfolded protein response (UPR). Reduced hepatic expression of the UPR regulator X-box binding protein 1 spliced (XBP1s) is associated with human nonalcoholic steatohepatitis (), and feeding mice a high-fat diet with fructose/sucrose causes progressive, fibrosing steatohepatitis. This study examines the role of XBP1 in nonalcoholic fatty liver injury and fatty -induced cell injury. Hepatocyte-specific Xbp1-deficient (Xbp1(-/-)) mice were fed a high-fat/sugar (HFS) diet for up to 16 wk. HFS-fed Xbp1(-/-) mice exhibited higher serum alanine aminotransferase levels compared with Xbp1(fl/fl) controls. RNA sequencing and Gene Ontogeny pathway analysis of hepatic mRNA revealed that apoptotic process, inflammatory response, and extracellular matrix structural constituent pathways had enhanced activation in HFS-fed Xbp1(-/-) mice. Liver histology demonstrated enhanced injury and fibrosis but less steatosis in the HFS-fed Xbp1(-/-) mice. Hepatic Col1a1 and Tgfβ1 gene expression, as well as Chop and phosphorylated JNK (p-JNK), were increased in Xbp1(-/-) compared with Xbp1(fl/fl) mice after HFS feeding. In vitro, stable XBP1-knockdown Huh7 cells (Huh7-KD) and scramble control cells (Huh7-SCR) were generated and treated with (PA) for 24 h. PA-treated Huh7-KD cells had increased cytotoxicity measured by lactate dehydrogenase release, apoptotic nuclei, and caspase3/7 activity assays compared with Huh7-SCR cells. CHOP and p-JNK expression was also increased in Huh7-KD cells following PA treatment. In conclusion, loss of XBP1 enhances injury in both in vivo and in vitro models of fatty liver injury. We speculate that hepatic XBP1 plays an important protective role in pathogenesis of .Copyright © 2015 the American Physiological Society.

Keyword: NASH

Macrophage-Specific Hypoxia-Inducible Factor-1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis.

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia-inducible factors (HIFs) have recently been identified as important regulators of immunity and inflammation. In nonalcoholic steatohepatitis (), HIF-1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF-1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF-1α levels in hepatic macrophages in methionine-choline-deficient (MCD) diet-fed mice and in macrophages of patients with compared with controls. The HIF-1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin-1, LC3-II, and p62 in both mouse and human macrophages. LysM HIF fl/fl mice, which have HIF-1α levels stabilized in macrophages, showed higher steatosis and liver inflammation compared with HIF fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated fatty , (PA), both induces HIF-1α and impairs autophagic flux in macrophages. Using small interfering RNA-mediated knock-down and overexpression of HIF-1α in macrophages, we demonstrated that PA impairs autophagy via HIF-1α. We found that HIF-1α mediates NF-κB activation and MCP-1 production and that HIF-1α-mediated impairment of macrophage autophagy increases IL-1β production, contributing to MCD diet-induced . Conclusion: impairs autophagy via HIF-1α activation in macrophages. HIF-1α and impaired autophagy are present in in vivo in mouse macrophages and in human blood monocytes. We identified that HIF-1α activation and decreased autophagic flux stimulate inflammation in macrophages through upregulation of NF-κB activation. These results suggest that macrophage activation in involves a complex interplay between HIF-1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet-induced .© 2018 by the American Association for the Study of Liver Diseases.

Keyword: NASH

Lumican, an extracellular matrix proteoglycan, is a novel requisite for hepatic fibrosis.

Lumican, an extracellular matrix proteoglycan was previously shown to be upregulated with increasing severity of nonalcoholic steatohepatitis (). Although lumican is involved in collagen fibrillogenesis in extra-hepatic tissues, little is known about the role of lumican in hepatic disease. We therefore determined lumican expression in etiologies other than clinical . Our results indicated that lumican is upregulated in clinical samples of hepatitis C virus infection, in experimental rodent models of chronic and acute liver injury and could additionally be induced in vitro in response to the pro-fibrotic cytokine transforming growth factor β1 (TGFβ1) and to lipotoxic . Together, these results suggested a role for lumican in hepatic fibrosis. To investigate the functional role of lumican in hepatic fibrosis, lumican null (Null) and wild-type (WT) littermates were administered carbon tetrachloride intra-peritoneally. Serum and liver tissue were analyzed for indices of liver injury, fibrosis, matrix turnover, and proliferation. Hepatic fibrosis was greatly reduced in null animals (P<0.05). Paradoxically, gene expression of fibrosis-related genes such as TGFβ1 and collagen 1 was numerically higher in null animals though statistically insignificant from WT animals. On the other hand, α smooth muscle actin expression (α-SMA), a marker for activated fibroblasts, the main contributors of collagen production was significantly higher (P<0.05) in null animals as compared with WT littermates. Among the matrix metalloproteases (MMP), MMP13 was significantly increased (P<0.05) in null animals. Ultra-structural imaging indicated differences in the organization and spatial distribution of hepatic collagen fibrils of null and WT mice. Cell proliferation was significantly increased (P<0.05) in null animals. We conclude that lumican is a prerequisite for hepatic fibrosis. The protective effect of lumican deficiency in hepatic fibrosis appears to be downstream of collagen production and mediated through the combined effects of impaired collagen fibrillogenesis, increased matrix turnover, and an enhanced proliferative response.

Keyword: NASH

Increased hepatic apoptosis in high-fat diet-induced in rats may be associated with downregulation of hepatic stimulator substance.

The mechanisms of progression from fatty liver to steatohepatitis and cirrhosis are not well elucidated. Hepatocellular apoptosis could be one of the key factors in the pathogenesis of non-alcoholic steatohepatitis (). Hepatic stimulator substance (HSS) protects liver cells from various toxins. We previously reported that HSS is critically important for the survival of hepatocytes due to its mitochondrial association. This study aims to investigate the relationship between HSS and hepatocellular apoptosis in vivo models of high-fat diet-induced and in vitro models of -induced hepatocyte injury. Sprague-Dawley rats were fed a high-fat diet for 8, 12 and 16\xa0weeks. Hepatic histological lesions, liver function and apoptosis were examined. HSS expression, in association with caspase-3 and cytochrome c leakage, which are both indicators of cell apoptosis, was measured. Results showed that a high-fat diet altered liver function and histology in a manner resembling . Hepatic protein and mRNA HSS expression was decreased as progressed. Meanwhile, cell apoptosis increased as result of caspase-3 activation and cytochrome c release, indicating that HSS might be involved in pathogenesis. Furthermore, in -induced hepatic cell damage, over-expression of HSS decreased cells apoptosis. In contrast, repression of HSS expression by siRNA increased cell apoptosis. In conclusion, these data imply that cell apoptosis contributes to the pathogenesis of , during which HSS expression is downregulated. Increasing HSS expression in hepatocytes may forestall cell apoptosis as result of fatty insult.

Keyword: NASH

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis () is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis ().For in vitro studies, (PA) and free fatty (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess . Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of , in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of .Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: NASH

Elovl6 promotes nonalcoholic steatohepatitis.

Nonalcoholic steatohepatitis () is associated with obesity and type 2 diabetes, and an increased risk for liver cirrhosis and cancer. ELOVL family member 6, elongation of very long chain fatty acids (Elovl6), is a microsomal enzyme that regulates the elongation of C12-16 saturated and monounsaturated fatty acids (FAs). We have shown previously that Elovl6 is a major target for sterol regulatory element binding proteins in the liver and that it plays a critical role in the development of obesity-induced insulin resistance by modifying FA composition. To further investigate the role of Elovl6 in the development of and its underlying mechanism, we used three independent mouse models with loss or gain of function of Elovl6, and human liver samples isolated from patients with . Our results demonstrate that (1) Elovl6 is a critical modulator for atherogenic high-fat diet-induced inflammation, oxidative stress, and fibrosis in the liver; (2) Elovl6 expression is positively correlated with severity of hepatosteatosis and liver injury in patients; and (3) deletion of Elovl6 reduces palmitate-induced activation of the NLR family pyrin domain-containing 3 inflammasome; this could be at least one of the underlying mechanisms by which Elovl6 modulates the progress of .Hepatic long-chain fatty composition is a novel determinant in development, and Elovl6 could be a potential therapeutic target for the prevention and treatment of .Copyright © 2012 American Association for the Study of Liver Diseases.

Keyword: NASH

Lysophosphatidylcholine as a death effector in the lipoapoptosis of hepatocytes.

The pathogenesis of nonalcoholic steatohepatitis () is unclear, despite epidemiological data implicating FFAs. We studied the pathogenesis of using lipoapoptosis models. (PA) induced classical apoptosis of hepatocytes. PA-induced lipoapoptosis was inhibited by acyl-CoA synthetase inhibitor but not by ceramide synthesis inhibitors, suggesting that conversion products other than ceramide are involved. Phospholipase A(2) (PLA(2)) inhibitors blocked PA-induced hepatocyte death, suggesting an important role for PLA(2) and its product lysophosphatidylcholine (LPC). Small interfering RNA for Ca(2+)-independent phospholipase A(2) (iPLA(2)) inhibited the lipoapoptosis of hepatocytes. PA increased LPC content, which was reversed by iPLA(2) inhibitors. Pertussis toxin or dominant-negative Galpha(i) mutant inhibited hepatocyte death by PA or LPC acting through G-protein-coupled receptor (GPCR)/Galpha(i). PA decreased cardiolipin content and induced mitochondrial potential loss and cytochrome c translocation. Oleic inhibited PA-induced hepatocyte death by diverting PA to triglyceride and decreasing LPC content, suggesting that FFAs lead to steatosis or lipoapoptosis according to the abundance of saturated/unsaturated FFAs. LPC administration induced hepatitis in vivo. LPC content was increased in the liver specimens from patients. These results demonstrate that LPC is a death effector in the lipoapoptosis of hepatocytes and suggest potential therapeutic values of PLA(2) inhibitors or GPCR/Galpha(i) inhibitors in .

Keyword: NASH

Treatment of cigarette smoke extract and condensate differentially potentiates -induced lipotoxicity and steatohepatitis in vitro.

Accumulative evidence showed that cigarette smoke (CS) detrimentally affects the pathogenesis of nonalcoholic steatohepatitis (). The purpose of this study was to evaluate the effects of CS extract (CSE) or total particulate matter (TPM) on the in vitro steatohepatitis model using mouse primary hepatocytes treated with (PA) or PA plus LPS. Increased hepatocellular damage was observed in PA-treated hepatocytes with TPM or CSE treatment, but increased triglyceride level was only observed in PA plus LPS-treated hepatocytes with a high concentration of TPM. Also, expression levels of steatohepatitis-related genes such as TNF-α, NOS 2, and SREBP-1c were significantly increased after treatment of TPM. To further demonstrate the role of Kupffer cells (KCs) after CS extracts treatment, trans-well co-culture system of hepatocytes and KCs was utilized. The levels of inflammatory cytokines and the ratios of Bax/Bcl-2 (apoptosis-related genes) were markedly increased in co-cultured hepatocytes after TPM or CSE treatment. Interestingly, KCs activation was augmented in KCs upon treatment with CSE or TPM. Overall, our findings indicate that in vitro treatment with CSE or TPM differentially contributes to the severity of steatohepatitis by modulating steatohepatitis-related lipotoxicity and inflammation, which might be caused by KCs activation with subsequent induction of hepatocytes apoptosis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: NASH

NLRP3 Deletion Inhibits the Non-alcoholic Steatohepatitis Development and Inflammation in Kupffer Cells Induced by .

The cleavage and secretion of pro-inflammatory cytokines IL-1β and IL-18 is regulated by NLRP3 (NACHT, LRR, and PYD domain-containing protein 3) inflammasome activation. Kupffer cells (KCs) are implicated in the pathogenesis of various liver diseases, such as non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, and liver fibrosis. However, the role of NLRP3 played in the non-alcoholic steatohepatitis () has yet to be evaluated. In the present study, methionine-choline-deficient (MCD) diet was used to establish the mice model. The expression levels of F4/80 and NLRP3 in liver tissues were evaluated, and the IL-1β and IL-18 in serum were also evaluated. KCs were isolated from wild-type (WT) mice and NLRP3 knockout (NLRP3) mice and then randomly divided into two groups: the control and (PA) groups. The expression levels of NLRP3, ASC, and caspase-1 in KCs were determined by RT-PCR, western blotting, and immunofluorescence. The levels of IL-1β and IL-18 in the supernatant (SN) of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). We found that KCs and NLRP3 play pro-inflammatory roles in the progression of , probably through secretions of IL-1β and IL-18 by KCs induced by PA. PA could act as a kind of damage-associated molecular patterns to elevate the messenger RNA and protein expression levels of NLRP3, ASC, and caspase-1 in KCs from WT mice. In the contrast, NLRP3 deletion could inhibit the NLRP3 inflammasome upregulation and activation in KCs induced by PA. Furthermore, the levels of pro-inflammatory cytokines IL-1β and IL-18 in the SN of KCs from WT mice were all elevated with the stimulation of PA, and the increase of these cytokines in the SN was blocked by NLRP3 deletion. In conclusion, our novel findings demonstrate that NLRP3 plays a pivotal role in development and pro-inflammatory cytokines IL-1β and IL-18 secretion induced by PA stimulation, and NLRP3 might be an effective potential target for the treatment of liver inflammatory diseases associated with NLRP3 inflammasome activation.

Keyword: NASH

Cathepsin B inhibition ameliorates the non-alcoholic steatohepatitis through suppressing caspase-1 activation.

Non-alcoholic fatty liver disease (NAFLD) has emerged as the most common chronic liver disease. NLRP3 inflammasome activation has been widely studied in the pathogenesis of NAFLD. Cathepsin B (CTSB) is a ubiquitous cysteine cathepsin, and the role of CTSB in the progression and development of NAFLD has received extensive concern. However, the exact roles of CTSB in the NAFLD development and NLRP3 inflammasome activation are yet to be evaluated. In the present study, we used methionine choline-deficient (MCD) diet to establish mice model. CTSB inhibitor (CA-074) was used to suppress the expression of CSTB. Expressions of CTSB and caspase-1 were evaluated by immunohistochemical staining. Serum IL-1β and IL-18 levels were also determined. was used to stimulate Kupffer cells (KCs), and protein expressions of CTSB, NLRP3, ASC (apoptosis-associated speck-like protein containing CARD), and caspase-1 in KCs were detected. The levels of IL-1β and IL-18 in the supernatant of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). Our results showed that CTSB inhibition improved the liver function and reduced hepatic inflammation and ballooning, and the levels of pro-inflammatory cytokines IL-1β and IL-18 were decreased. The expressions of CTSB and caspase-1 in liver tissues were increased in the group. In in vitro experiments, PA stimulation could increase the expressions of CTSB and NLRP3 inflammasome in KCs, and CTSB inhibition downregulated the expression of NLRP3 inflammasome in KCs, when challenged by PA. Moreover, CTSB inhibition effectively suppressed the expression and activity of caspase-1 and subsequently secretions of IL-1β and IL-18. Collectively, these results suggest that CTSB inhibition limits NLRP3 inflammasome-dependent formation through regulating the expression and activity of caspase-1, thus providing a novel anti-inflammatory signal pathway for the therapy of NAFLD.

Keyword: NASH

Degradation of splicing factor SRSF3 contributes to progressive liver disease.

Serine rich splicing factor 3 (SRSF3) plays a critical role in liver function and its loss promotes chronic liver damage and regeneration. As a consequence, genetic deletion of SRSF3 in hepatocytes caused progressive liver disease and ultimately led to hepatocellular carcinoma. Here we show that SRSF3 is decreased in human liver samples with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (), or cirrhosis that was associated with alterations in RNA splicing of known SRSF3 target genes. Hepatic SRSF3 expression was similarly decreased and RNA splicing dysregulated in mouse models of NAFLD and . We showed that -induced oxidative stress caused conjugation of the ubiquitin like NEDD8 protein to SRSF3 and proteasome mediated degradation. SRSF3 was selectively neddylated at lysine11 and mutation of this residue (SRSF3-K11R) was sufficient to prevent both SRSF3 degradation and alterations in RNA splicing. Finally prevention of SRSF3 degradation in vivo partially protected mice from hepatic steatosis, fibrosis and inflammation. These results highlight a neddylation-dependent mechanism regulating gene expression in the liver that is disrupted in early metabolic liver disease and may contribute to the progression to , cirrhosis and ultimately hepatocellular carcinoma.

Keyword: NASH

Cellular glutathione in fatty liver in vitro models.

The range of non-alcoholic fatty liver disease (NAFLD) includes simple hepatic steatosis, the inflammatory non-alcoholic steatohepatitis (), fibrosis and cirrhosis. The accumulation of specific lipids in hepatocytes has been associated with oxidative stress and progression of the disease. Elevated serum free fatty acids and hepatocyte lipotoxicity can be studied in an in vitro cellular model. For this purpose, we cultured the human liver cell line, HepG2/C3A, in medium supplemented with increasing amounts of oleic (C18:1) and evaluated oxidative stress by measuring the content of the cellular antioxidant, glutathione (GSH). We observed a dose-dependent steatosis, as determined by Nile Red staining, with concurrent increases of GSH; similar findings were also observed in cultured human hepatocytes. Cells cultured with (C16:0) or the combination oleic/ acids (2:1 ratio) also exhibited a dose-dependent increase of GSH; however -supplemented cultures did not sustain the GSH increase after 24h. We also detected an increase in the formation of lipid peroxides (LPO) indicating that the increase of GSH was a cellular mechanism that may be related to the high exposure of fatty acids. The results of this in vitro study suggest an antioxidant response against fat overloading and indicate potential differences in response to specific fatty -induced hepatic steatosis and associated lipotoxicity.Published by Elsevier Ltd.

Keyword: NASH

Gut microbiota-mediated generation of saturated fatty acids elicits inflammation in the liver in murine high-fat diet-induced steatohepatitis.

The gut microbiota plays crucial roles in the development of non-alcoholic steatohepatitis (). However, the precise mechanisms by which alterations of the gut microbiota and its metabolism contributing to the pathogenesis of are not yet fully elucidated.Mice were fed with a recently reported new class of high-fat diet (HFD), steatohepatitis-inducing HFD (STHD)-01 for 9\xa0weeks. The composition of the gut microbiota was analyzed by T-RFLP. Luminal metabolome was analyzed using capillary electrophoresis and liquid chromatography time-of-flight mass spectrometry (CE- and LC-TOFMS).Mice fed the STHD-01 developed -like pathology within a short period. Treatment with antibiotics prevented the development of by STHD-01. The composition of the gut microbiota and its metabolic activities were markedly perturbed in the STHD-01-fed mice, and antibiotic administration normalized these changes. We identified that long-chain saturated fatty and n-6 fatty metabolic pathways were significantly altered by STHD-01. Of note, the changes in gut lipidome caused by STHD-01 were mediated by gut microbiota, as the depletion of the gut microbiota could reverse the perturbation of these metabolic pathways. A saturated long-chain fatty , , which accumulated in the STHD-01 group, activated liver macrophages and promoted TNF-α expression.Lipid metabolism by the gut microbiota, particularly the saturation of fatty acids, affects fat accumulation in the liver and subsequent liver inflammation in .

Keyword: NASH

Hepatic vagus nerve regulates Kupffer cell activation via α7 nicotinic acetylcholine receptor in nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease ranges from simple steatosis to nonalcoholic steatohepatitis (). Kupffer cells play a central role in promoting hepatic inflammation, which leads to the development of . We investigated the anti-inflammatory effect of hepatic vagus-mediated stimulation of the α7 nicotinic acetylcholine receptor (α7nAChR) on Kupffer cells in pathogenesis.Wild-type (WT) mice undergoing hepatic vagotomy (HV) were fed a methionine- and choline-deficient (MCD) diet for 1\xa0week. α7nAChR knockout (α7KO) chimeric mice were generated by transplanting α7KO bone marrow cells into irradiated and Kupffer cell-deleted WT recipients. Kupffer cells were isolated from WT mice and treated with α7nAChR agonist under stimulation by lipopolysaccharide and/or .HV aggravated MCD diet-induced in both steatosis and inflammation. The hepatic inflammatory response, including the upregulation of tumor necrosis factor alpha (TNFα), interleukin (IL)-12, and monocyte chemoattractant protein 1 (MCP-1), was accelerated in HV mice, accompanied by the downregulation of PPARα pathway genes. Kupffer cells were highly activated via the phosphorylation and nuclear translocation of nuclear factor-kappa B (NF-κB) in MCD diet-fed HV mice. The α7nAchR agonist suppressed the inflammatory response of primary Kupffer cells induced by lipopolysaccharide and by attenuating the NF-κB cascade. α7KO chimeric mice fed an MCD diet for 1\xa0week developed advanced with highly activated Kupffer cells. The hepatic expression of TNFα, IL-12, and MCP-1 was upregulated in α7KO chimeric mice, accompanied by abnormal lipid metabolism.Hepatic vagus activity regulates the inflammatory response of Kupffer cells via α7nAChR in development.

Keyword: NASH

Inhibition of MD2-dependent inflammation attenuates the progression of non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) can progress to the more serious non-alcoholic steatohepatitis (), characterized by inflammatory injury and fibrosis. The pathogenic basis of NAFLD progressing to is currently unknown, but growing evidence suggests MD2 (myeloid differentiation factor 2), an accessory protein of TLR4, is an important signalling component contributing to this disease. We evaluated the effectiveness of the specific MD2 inhibitor, L6H21, in reducing inflammatory liver injury in a relevant high-fat diet (HFD) mouse model of and in the (PA)-stimulated human liver cell line (HepG2). For study, genetic knockout (MD2 ) mice were fed a HFD or control diet for 24 weeks, or wild-type mice placed on a similar diet regimen and treated with L6H21 for the last 8 or 16 weeks. Results indicated that MD2 inhibition with L6H21 was as effective as MD2 knockout in preventing the HFD-induced hepatic lipid accumulation, pro-fibrotic changes and expression of pro-inflammatory molecules. Direct challenge of HepG2 with PA (200 μM) increased MD2-TLR4 complex formation and expression of pro-inflammatory and pro-fibrotic genes and L6H21 pre-treatment prevented these PA-induced responses. Interestingly, MD2 knockout or L6H21 increased expression of the anti-inflammatory molecule, PPARγ, in liver tissue and the liver cell line. Our results provide further evidence for the critical role of MD2 in the development of and conclude that MD2 could be a potential therapeutic target for NAFLD/ treatment. Moreover, the small molecule MD2 inhibitor, L6H21, was an effective and selective investigative agent for future mechanistic studies of MD2.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: NASH

Metabolic pathways promoting intrahepatic fatty accumulation in methionine and choline deficiency: implications for the pathogenesis of steatohepatitis.

The pathological mechanisms that distinguish simple steatosis from steatohepatitis (or , with consequent risk of cirrhosis and hepatocellular cancer) remain incompletely defined. Whereas both a methionine- and choline-deficient diet (MCDD) and a choline-deficient diet (CDD) lead to hepatic triglyceride accumulation, MCDD alone is associated with hepatic insulin resistance and inflammation (steatohepatitis). We used metabolic tracer techniques, including stable isotope ([¹³C₄]palmitate) dilution and mass isotopomer distribution analysis (MIDA) of [¹³C₂]acetate, to define differences in intrahepatic fatty metabolism that could explain the contrasting effect of MCDD and CDD on in C57Bl6 mice. Compared with control-supplemented (CS) diet, liver triglyceride pool sizes were similarly elevated in CDD and MCDD groups (24.37 ± 2.4, 45.94 ± 3.9, and 43.30 ± 3.5 μmol/liver for CS, CDD, and MCDD, respectively), but intrahepatic neutrophil infiltration and plasma alanine aminotransferase (31 ± 3, 48 ± 4, 231 ± 79 U/l, P < 0.05) were elevated only in MCDD mice. However, despite loss of peripheral fat in MCDD mice, neither the rate of appearance of palmitate (27.2 ± 3.5, 26.3 ± 2.3, and 28.3 ± 3.5 μmol·kg⁻¹·min⁻¹) nor the contribution of circulating fatty acids to the liver triglyceride pool differed between groups. Unlike CDD, MCDD had a defect in hepatic triglyceride export that was confirmed using intravenous tyloxapol (142 ± 21, 122 ± 15, and 80 ± 7 mg·kg⁻¹·h⁻¹, P < 0.05). Moreover, hepatic de novo lipogenesis was significantly elevated in the MCDD group only (1.4 ± 0.3, 2.3 ± 0.4, and 3.4 ± 0.4 μmol/day, P < 0.01). These findings suggest that important alterations in hepatic fatty metabolism may promote the development of steatohepatitis. Similar mechanisms may predispose to hepatocyte damage in human .

Keyword: NASH

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of , obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe liver inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: NASH

Effect of intracellular lipid accumulation in a new model of non-alcoholic fatty liver disease.

In vitro exposure of liver cells to high concentrations of free fatty acids (FFA) results in fat overload which promotes inflammatory and fibrogenic response similar to those observed in patients with Non-Alcoholic Fatty Liver Disease (NAFLD) and Non-Alcoholic Steatohepatitis (). Since the mechanisms of this event have not been fully characterized, we aimed to analyze the fibrogenic stimuli in a new in vitro model of .HuH7 cells were cultured for 24 h in an enriched medium containing bovine serum albumin and increasing concentrations of and oleic at a molar ratio of 1:2 ( and oleic , respectively). Cytotoxic effect, apoptosis, oxidative stress, and production of inflammatory and fibrogenic cytokines were measured.FFA induces a significant increment in the intracellular content of lipid droplets. The gene expression of interleukin-6, interleukin-8 and tumor necrosis factor alpha was significantly increased. The protein level of interleukin-8 was also increased. Intracellular lipid accumulation was associated to a significant up-regulation in the gene expression of transforming growth factor beta 1, alpha 2 macroglobulin, vascular endothelial growth factor A, connective tissue growth factor, insulin-like growth factor 2, thrombospondin 1. Flow cytometry analysis demonstrated a significant increment of early apoptosis and production of reactive oxygen species.The exposure of hepatocytes to fatty acids elicits inflammation, increase of oxidative stress, apoptosis and production of fibrogenic cytokines. These data support a primary role of FFA in the pathogenesis of NAFLD and .

Keyword: NASH

Downregulation of microRNA-451 in non-alcoholic steatohepatitis inhibits fatty -induced proinflammatory cytokine production through the AMPK/AKT pathway.

Mechanisms associated with the progression of non-alcoholic fatty liver disease (NAFLD) remain unclear. We attempted to identify the pattern of altered gene expression at different time points in a high fat diet (HFD)-induced NAFLD mouse model. The early up-regulated genes are mainly involved in the innate immune responses, while the late up-regulated genes represent the inflammation processes. Although recent studies have shown that microRNAs play important roles in hepatic metabolic functions, the pivotal role of microRNAs in the progression of NAFLD is not fully understood. We investigated the functions of miR-451, which was identified as a target gene in the inflammatory process in NAFLD. miR-451 expression was significantly decreased in the palmitate (PA)-exposed HepG2 cells and in liver tissues of HFD-induced non-alcoholic steatohepatitis () mice. Its decreased expressions were also observed in liver specimens of patients. In vitro analysis of the effect of miR-451 on proinflammatory cytokine provided evidence for negative regulation of PA-induced interleukin (IL)-8 and tumor necrosis factor-alpha (TNF-α) production. Furthermore, miR-451 over-expression inhibited translocation of the PA-induced NF-κB p65 subunit into the nucleus. Our result showed that Cab39 is a direct target of miRNA-451 in steatotic cells. Further study showed that AMPK activated through Cab39 inhibits NF-κB transactivation induced in steatotic HepG2 cells. miR-451 over-expression in steatotic cells significantly suppressed PA-induced inflammatory cytokine. These results provide new insights into the negative regulation of miR-451 in fatty -induced inflammation via the AMPK/AKT pathway and demonstrate potential therapeutic applications for miR-451 in preventing the progression from simple steatosis to severely advanced liver disease.Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Keyword: NASH

plus Synergistically Ameliorates Nonalcoholic Steatohepatitis in HepG2 Cells.

The combination of and radix is frequently prescribed for liver diseases in TKM. However, the synergic effects of the two herbs on nonalcoholic steatohepatitis () have not yet been studied. Therefore, we investigated the anti- effects of the water extract of (AI), radix (CL), and combination of the two herbs (ACE). Hepatic steatosis and were induced in HepG2 cells by treatment with (PA, for 6\u2009h) with/without pretreatment of ACE (25 or 50\u2009g/mL), AI (50 or 100\u2009g/mL), CL (50 or 100\u2009g/mL), curcumin (5\u2009g/mL), or scopoletin (5\u2009g/mL). The PA treatment (200\u2009M) drastically altered intracellular triglyceride levels, total cholesterol, and expression levels of genes related to lipid metabolism (CD36, SREBP1c, PPAR-, and PPAR-), whereas pretreatment with ACE significantly attenuated these alterations. ACE also protected HepG2 cells from PA- (300\u2009M-) induced endoplasmic reticulum (ER) stress and apoptosis and attenuated the related key molecules including GRP78, eIF2, and CHOP, respectively. In conclusion, we found synergic effects of and on , supporting the clinical potential for fatty liver disorders. In addition, modulation of ER stress-relative molecules would be involved in its underlying mechanism.

Keyword: NASH

Serum C16:1n7/C16:0 ratio as a diagnostic marker for non-alcoholic steatohepatitis.

Accurate diagnosis of non-alcoholic steatohepatitis () from non-alcoholic fatty liver disease (NAFLD) is clinically important. Therefore, there is a need for easier ways of diagnosing . In this study, we investigated the serum fatty composition and evaluated the possibility of using the serum fatty composition as a diagnostic marker of .The subjects were 78 NAFLD patients (non-alcoholic fatty liver [NAFL]: 30, : 48) and 24 healthy individuals. Fatty acids extracted from the liver tissue and serum were identified and quantified by gas chromatography. In addition, we evaluated the relationship between serum and liver tissue fatty composition, patient background, and liver histology. The diagnostic performance of was evaluated by calculating the area under the receiver operating characteristic (AUROC).The results of the fatty analysis showed the C16:1n7/C16:0 ratio to have the strongest correlation between serum and liver tissue (r\xa0=\xa00.865, P\xa0<\xa00.0001). The serum C16:1n7/C16:0 ratio in the group was higher compared with that in the NAFL group (P\xa0=\xa00.0007). Evaluation of the association of the serum C16:1n7/C16:0 ratio with liver histology revealed significant correlation with lobular inflammation score, ballooning score, and fibrosis score. The AUROC for predicting in all NAFLD patients was 0.7097. The AUROC was nearly equivalent even when the study subjects were restricted to patients with a fibrosis score\xa0≤\xa02 only (AUROC 0.6917).Measuring the serum C16:1n7/C16:0 ratio may be an effective non-invasive method for diagnosing , particularly in its early stages.© 2019 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Keyword: NASH

Mixed Lineage Kinase 3 Mediates the Induction of CXCL10 by a STAT1-Dependent Mechanism During Hepatocyte Lipotoxicity.

Saturated fatty acids (SFA) and their toxic metabolites contribute to hepatocyte lipotoxicity in nonalcoholic steatohepatitis (). We previously reported that hepatocytes, under lipotoxic stress, express the potent macrophage chemotactic ligand C-X-C motif chemokine 10 (CXCL10), and release CXCL10-enriched extracellular vesicles (EV) by a mixed lineage kinase (MLK) 3-dependent mechanism. In the current study, we sought to examine the signaling pathway responsible for CXCL10 induction during hepatocyte lipotoxicity. Here, we demonstrate a role for signal transducer and activator of transcription (STAT) 1 in regulating CXCL10 expression. Huh7 and HepG2 cells were treated with lysophosphatidylcholine (LPC), the toxic metabolite of the SFA palmitate. In LPC-treated hepatocytes, CXCL10 induction is mediated by a mitogen activated protein kinase (MAPK) signaling cascade consisting of a relay kinase module of MLK3, MKK3/6, and p38. P38 in turn induces STAT1 Ser727 phosphorylation and CXCL10 upregulation in hepatocytes, which is reduced by genetic or pharmacological inhibition of this MAPK signaling cascade. The binding and activity of STAT1 at the CXCL10 gene promoter were identified by chromatin immunoprecipitation and luciferase gene expression assays. Promoter activation was attenuated by MLK3/STAT1 inhibition or by deletion of the consensus STAT1 binding sites within the CXCL10 promoter. In lipotoxic hepatocytes, MLK3 activates a MAPK signaling cascade, resulting in the activating phosphorylation of STAT1, and CXCL10 transcriptional upregulation. Hence, this kinase relay module and/or STAT1 inhibition may serve as a therapeutic target to reduce CXCL10 release, thereby attenuating pathogenesis. J. Cell. Biochem. 118: 3249-3259, 2017.© 2017 Wiley Periodicals, Inc.

Keyword: NASH

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic fatty liver disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (), cirrhosis, and cancer. The role of different types of fatty acids in driving the early progression of NAFL to is not understood. Lipid overload causing lipotoxicity and inflammation has been considered as an essential pathogenic factor. To correlate the lipid profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced lipid overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced inflammation and cell death, whereas C16:1 induced significant lipid droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. Lipid profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific fatty acids that affect lipid metabolism and cause lipotoxicity or lipid droplet formation. It indicates that not only saturation or monounsaturation of fatty acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of lipid metabolism and the heterogeneous development of NAFLD.

Keyword: NASH

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis () associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced , while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: NASH

Dynamic alterations in the gut microbiota and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut microbiota and metabolism during nonalcoholic steatohepatitis () development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut microbiota dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut microbiota and metabolome.

Keyword: NASH

Lipotoxicity and the gut-liver axis in pathogenesis.

The pathogenesis of non-alcoholic fatty liver disease, particularly the mechanisms whereby a minority of patients develop a more severe phenotype characterised by hepatocellular damage, inflammation, and fibrosis is still incompletely understood. Herein, we discuss two pivotal aspects of the pathogenesis of . We first analyse the initial mechanisms responsible for hepatocellular damage and inflammation, which derive from the toxic effects of excess lipids. Accumulating data indicate that the total amount of triglycerides stored in hepatocytes is not the major determinant of lipotoxicity, and that specific lipid classes act as damaging agents on liver cells. In particular, the role of free fatty acids such as , cholesterol, lysophosphatidylcholine and ceramides has recently emerged. These lipotoxic agents affect the cell behaviour via multiple mechanisms, including activation of signalling cascades and death receptors, endoplasmic reticulum stress, modification of mitochondrial function, and oxidative stress. In the second part of this review, the cellular and molecular players involved in the cross-talk between the gut and the liver are considered. These include modifications to the microbiota, which provide signals through the intestine and bacterial products, as well as hormones produced in the bowel that affect metabolism at different levels including the liver. Finally, the activation of nuclear receptors by bile acids is analysed.Copyright © 2017. Published by Elsevier B.V.

Keyword: NASH

Cellular cholesterol accumulation modulates high fat high sucrose (HFHS) diet-induced ER stress and hepatic inflammasome activation in the development of non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (), is the form of non-alcoholic fatty liver disease posing risk to progress into serious long term complications. Human and pre-clinical models implicate cellular cholesterol dysregulation playing important role in its development. Mouse model studies suggest synergism between dietary cholesterol and fat in contributing to but the mechanisms remain poorly understood. Our laboratory previously reported the primary importance of hepatic endoplasmic reticulum cholesterol (ER-Chol) in regulating hepatic ER stress by comparing the responses of wild type, Ldlr-/-xLcat+/+ and Ldlr-/-xLcat-/- mice, to a 2% high cholesterol diet (HCD). Here we further investigated the roles of ER-Chol and ER stress in HFHS diet-induced using the same strains. With HFHS diet feeding, both WT and Ldlr-/-xLcat+/+ accumulate ER-Chol in association with ER stress and inflammasome activation but the Ldlr-/-xLcat-/- mice are protected. By contrast, all three strains accumulate cholesterol crystal, in correlation with ER-Chol, albeit less so in Ldlr-/-xLcat-/- mice. By comparison, HCD feeding per se (i) is sufficient to promote steatosis and activate inflammasomes, and (ii) results in dramatic accumulation of cholesterol crystal which is linked to inflammasome activation in Ldlr-/-xLcat-/- mice, independent of ER-Chol. Our data suggest that both dietary fat and cholesterol each independently promote steatosis, cholesterol crystal accumulation and inflammasome activation through distinct but complementary pathways. In vitro studies using palmitate-induced hepatic steatosis in HepG2 cells confirm the key roles by cellular cholesterol in the induction of steatosis and inflammasome activations. These novel findings provide opportunities for exploring a cellular cholesterol-focused strategy for treatment of .Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: NASH

Lipoapoptosis induced by saturated free fatty acids stimulates monocyte migration: a novel role for Pannexin1 in liver cells.

Recruitment of monocytes in the liver is a key pathogenic feature of hepatic inflammation in nonalcoholic steatohepatitis (), but the mechanisms involved are poorly understood. Here, we studied migration of human monocytes in response to supernatants obtained from liver cells after inducing lipoapoptosis with saturated free fatty acids (FFA). Lipoapoptotic supernatants stimulated monocyte migration with the magnitude similar to a monocyte chemoattractant protein, CCL2 (MCP-1). Inhibition of c-Jun NH2-terminal kinase (JNK) in liver cells with SP600125 blocked migration of monocytes in a dose-dependent manner, indicating that JNK stimulates release of chemoattractants in lipoapoptosis. Notably, treatment of supernatants with Apyrase to remove ATP potently inhibited migration of THP-1 monocytes and partially blocked migration of primary human monocytes. Inhibition of the CCL2 receptor (CCR2) on THP-1 monocytes with RS102895, a specific CCR2 inhibitor, did not block migration induced by lipoapoptotic supernatants. Consistent with these findings, lipoapoptosis stimulated pathophysiological extracellular ATP (eATP) release that increased supernatant eATP concentration from 5 to ~60 nM. Importantly, inhibition of Panx1 expression in liver cells with short hairpin RNA (shRNA) decreased supernatant eATP concentration and inhibited monocyte migration, indicating that monocyte migration is mediated in part by Panx1-dependent eATP release. Moreover, JNK inhibition decreased supernatant eATP concentration and inhibited Pannexin1 activation, as determined by YoPro-1 uptake in liver cells in a dose-dependent manner. These results suggest that JNK regulates activation of Panx1 channels, and provide evidence that Pannexin1-dependent pathophysiological eATP release in lipoapoptosis is capable of stimulating migration of human monocytes, and may participate in the recruitment of monocytes in chronic liver injury induced by saturated FFA.

Keyword: NASH

Attenuated lipotoxicity and apoptosis is linked to exogenous and endogenous augmenter of liver regeneration by different pathways.

Nonalcoholic fatty liver disease (NAFLD) covers a spectrum from simple steatosis to nonalcoholic steatohepatitis () and cirrhosis. Free fatty acids (FFA) induce steatosis and lipo-toxicity and correlate with severity of NAFLD. In this study we aimed to investigate the role of exogenous and endogenous ALR (augmenter of liver regeneration) for FFA induced ER (endoplasmatic reticulum) -stress and lipoapoptosis. Primary human hepatocytes or hepatoma cells either treated with recombinant human ALR (rhALR, 15kDa) or expressing short form ALR (sfALR, 15kDa) were incubated with (PA) and analyzed for lipo-toxicity, -apoptosis, activation of ER-stress response pathways, triacylglycerides (TAG), mRNA and protein expression of lipid metabolizing genes. Both, exogenous rhALR and cytosolic sfALR reduced PA induced caspase 3 activity and Bax protein expression and therefore lipotoxicity. Endogenous sfALR but not rhALR treatment lowered TAG levels, diminished activation of ER-stress mediators C-Jun N-terminal kinase (JNK), X-box binding protein-1 (XBP1) and proapoptotic transcription factor C/EBP-homologous protein (CHOP), and reduced death receptor 5 protein expression. Cellular ALR exerts its lipid lowering and anti-apoptotic actions by enhancing FABP1, which binds toxic FFA, increasing mitochondrial β-oxidation by elevating the mitochondrial FFA transporter CPT1α, and decreasing ELOVL6, which delivers toxic FFA metabolites. We found reduced hepatic mRNA levels of ALR in a high fat diet mouse model, and of ALR and FOXA2, a transcription factor inducing ALR expression, in human steatotic as well as liver samples, which may explain increased lipid deposition and reduced β-oxidation in patients. Present study shows that exogenous and endogenous ALR reduce PA induced lipoapoptosis. Furthermore, cytosolic sfALR changes mRNA and protein expression of genes regulating lipid metabolism, reduces ER-stress finally impeding progression of .

Keyword: NASH

Necroptosis is a key pathogenic event in human and experimental murine models of non-alcoholic steatohepatitis.

Hepatocyte cell death, inflammation and oxidative stress constitute key pathogenic mechanisms underlying non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the role of necroptosis in human and experimental NAFLD and its association with tumour necrosis factor α (TNF-α) and oxidative stress. Serum markers of necrosis, liver receptor-interacting protein 3 (RIP3) and phosphorylated mixed lineage kinase domain-like (MLKL) were evaluated in control individuals and patients with NAFLD. C57BL/6 wild-type (WT) or RIP3-deficient (RIP3(-/-)) mice were fed a high-fat choline-deficient (HFCD) or methionine and choline-deficient (MCD) diet, with subsequent histological and biochemical analysis of hepatic damage. In primary murine hepatocytes, necroptosis and oxidative stress were also assessed after necrostatin-1 (Nec-1) treatment or RIP3 silencing. We show that circulating markers of necrosis and TNF-α, as well as liver RIP3 and MLKL phosphorylation were increased in NAFLD. Likewise, RIP3 and MLKL protein levels and TNF-α expression were increased in the liver of HFCD and MCD diet-fed mice. Moreover, RIP3 and MLKL sequestration in the insoluble protein fraction of (non-alcoholic steatohepatitis) mice liver lysates represented an early event during stetatohepatitis progression. Functional studies in primary murine hepatocytes established the association between TNF-α-induced RIP3 expression, activation of necroptosis and oxidative stress. Strikingly, RIP3 deficiency attenuated MCD diet-induced liver injury, steatosis, inflammation, fibrosis and oxidative stress. In conclusion, necroptosis is increased in the liver of NAFLD patients and in experimental models of . Further, TNF-α triggers RIP3-dependent oxidative stress during hepatocyte necroptosis. As such, targeting necroptosis appears to arrest or at least impair NAFLD progression.© 2015 Authors; published by Portland Press Limited.

Keyword: NASH

Lipidomic-based investigation into the regulatory effect of Schisandrin B on level in non-alcoholic steatotic livers.

Schisandrin B (SchB) is one of the most abundant bioactive dibenzocyclooctadiene derivatives found in the fruit of Schisandra chinensis. Here, we investigated the potential therapeutic effects of SchB on non-alcoholic fatty-liver disease (NAFLD). In lipidomic study, ingenuity pathway analysis highlighted palmitate biosynthesis metabolic pathway in the liver samples of SchB-treated high-fat-diet-fed mice. Further experiments showed that the SchB treatment reduced expression and activity of fatty synthase, expressions of hepatic mature sterol regulatory element binding protein-1 and tumor necrosis factor-α, and hepatic level of which is known to promote progression of steatosis to steatohepatitis. Furthermore, the treatment also reduced hepatic fibrosis, activated nuclear factor-erythroid-2-related factor-2 which is known to attenuate the progression of -related fibrosis. Interestingly, in fasting mice, a single high-dose SchB induced transient lipolysis and increased the expressions of adipose triglyceride lipase and phospho-hormone sensitive lipase. The treatment also increased plasma cholesterol levels and 3-hydroxy-3-methylglutaryl-CoA reductase activity, reduced the hepatic low-density-lipoprotein receptor expression in these mice. Our data not only suggest SchB is a potential therapeutic agent for NAFLD, but also provided important information for a safe consumption of SchB because SchB overdosed under fasting condition will have adverse effects on lipid metabolism.

Keyword: NASH

induces autophagy in hepatocytes via JNK2 activation.

Free fatty -induced lipotoxicity plays a crucial role in the progression of nonalcoholic fatty liver disease (NAFLD). In the present study we investigated the effects of a high-fat diet and free fatty acids on the autophagic process in hepatocytes in vivo and in vitro and the underlying mechanisms.LC3-II expression, a hallmark of autophagic flux, was detected in liver specimens from patients with non-alcoholic steatohepatitis () as well as in the livers of C57BL/6 mice fed a high-fat diet (HFD) up to 16 weeks. LC3-II expression was also analyzed in human SMMC-7721 and HepG2 hepatoma cells exposed to (PA), a saturated fatty . PA-induced apoptosis was detected by Annexin V staining and specific cleavage of PARP in the presence and absence of different agents.LC3-II expression was markedly increased in human and in liver tissues of HFD-fed mice. Treatment of SMMC-7721 cells with PA increased LC3-II expression in time- and dose-dependent manners, whereas the unsaturated fatty oleic had no effect. Inhibition of autophagy with 3MA sensitized SMMC-7721 cells to PA-induced apoptosis, whereas activation of autophagy by rapamycin attenuated PA-induced PARP cleavage. The autophagy-associated proteins Beclin1 and Atg5 were essential for PA-induced autophagy in SMMC-7721 cells. Moreover, pretreatment with SP600125, an inhibitor of JNK, effectively abrogated PA-mediated autophagy and apoptosis. Specific knockdown of JNK2, but not JNK1, in SMMC-7721 cells significantly suppressed PA-induced autophagy and enhanced its pro-apoptotic activity; whereas specific knockdown of JNK1 had the converse effect. Similar results were obtained when HepG2 cells were tested.JNK1 promotes PA-induced lipoapoptosis, whereas JNK2 activates pro-survival autophagy and inhibits PA lipotoxicity. Our results suggest that modulation of autophagy may have therapeutic benefits in the treatment of lipid-related metabolic diseases.

Keyword: NASH

Hepatic steatosis causes induction of the chemokine RANTES in the absence of significant hepatic inflammation.

Nonalcoholic fatty liver disease (NAFLD) encompasses a spectrum ranging from simple steatosis to cirrhosis. Hepatocellular lipid accumulation is a hallmark of both nonalcoholic steatosis and steatohepatitis (). The latter develops upon pro-inflammatory cell infiltration and is widely considered as the first relevant pathophysiological step in NAFLD-progression. The chemokine CCL5/RANTES plays an important role in the progression of hepatic inflammation and fibrosis. We here aimed to investigate its expression in NAFLD. Incubation of primary human hepatocytes with induced a dose-dependent lipid accumulation, and corresponding dose-dependent RANTES induction in vitro. Furthermore, we observed significantly elevated hepatic RANTES expression in a dietary model of NAFLD, in which mice were fed a high-fat diet for 12 weeks. This diet induced significant hepatic steatosis but only minimal inflammation. In contrast to the liver, RANTES expression was not induced in visceral adipose tissue of the group fed with high-fat diet. Finally, RANTES serum levels were elevated in patients with ultrasound-diagnosed NAFLD. In conclusion, our data indicate hepatocytes as cellular source of elevated hepatic as well as circulating RANTES levels in response to hepatic steatosis. Noteworthy, upregulation of RANTES in response to lipid accumulation occurs in the absence of relevant inflammation, which further indicates that hepatic steatosis per se has pathophysiological relevance and should not be considered as benign.

Keyword: NASH

The plasma lipidomic signature of nonalcoholic steatohepatitis.

Specific alterations in hepatic lipid composition characterize the spectrum of nonalcoholic fatty liver disease (NAFLD), which extends from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (). However, the plasma lipidome of NAFLD and whether has a distinct plasma lipidomic signature are unknown. A comprehensive analysis of plasma lipids and eicosanoid metabolites quantified by mass spectrometry was performed in NAFL (n = 25) and (n = 50) subjects and compared with lean normal controls (n = 50). The key findings include significantly increased total plasma monounsaturated fatty acids driven by palmitoleic (16:1 n7) and oleic (18:1 n9) acids content (P < 0.01 for both acids in both NAFL and ). The levels of palmitoleic , oleic , and palmitoleic to (16:0) ratio were significantly increased in NAFLD across multiple lipid classes. Linoleic (8:2n6) was decreased (P < 0.05), with a concomitant increase in gamma-linolenic (18:3n6) and dihomo gamma-linolenic (20:3n6) acids in both NAFL and (P < 0.001 for most lipid classes). The docosahexanoic (22:6 n3) to docosapentenoic (22:5n3) ratio was significantly decreased within phosphatidylcholine (PC), and phosphatidylethanolamine (PE) pools, which was most marked in subjects (P < 0.01 for PC and P < 0.001 for PE). The total plasmalogen levels were significantly decreased in compared with controls (P < 0.05). A stepwise increase in lipoxygenase (LOX) metabolites 5(S)-hydroxyeicosatetraenoic (5-HETE), 8-HETE, and 15-HETE characterized progression from normal to NAFL to . The level of 11-HETE, a nonenzymatic oxidation product of arachidonic (20:4) , was significantly increased in only.Although increased lipogenesis, desaturases, and LOX activities characterize NAFL and , impaired peroxisomal polyunsaturated fatty (PUFA) metabolism and nonenzymatic oxidation is associated with progression to .

Keyword: NASH

Myristic potentiates -induced lipotoxicity and steatohepatitis associated with lipodystrophy by sustaning de novo ceramide synthesis.

(PA) induces hepatocyte apoptosis and fuels de novo ceramide synthesis in the endoplasmic reticulum (ER). Myristic (MA), a free fatty highly abundant in copra/palmist oils, is a predictor of nonalcoholic steatohepatitis () and stimulates ceramide synthesis. Here we investigated the synergism between MA and PA in ceramide synthesis, ER stress, lipotoxicity and . Unlike PA, MA is not lipotoxic but potentiated PA-mediated lipoapoptosis, ER stress, caspase-3 activation and cytochrome c release in primary mouse hepatocytes (PMH). Moreover, MA kinetically sustained PA-induced total ceramide content by stimulating dehydroceramide desaturase and switched the ceramide profile from decreased to increased ceramide 14:0/ceramide16:0, without changing medium and long-chain ceramide species. PMH were more sensitive to equimolar ceramide14:0/ceramide16:0 exposure, which mimics the outcome of PA plus MA treatment on ceramide homeostasis, than to either ceramide alone. Treatment with myriocin to inhibit ceramide synthesis and tauroursodeoxycholic to prevent ER stress ameliorated PA plus MA induced apoptosis, similar to the protection afforded by the antioxidant BHA, the pan-caspase inhibitor z-VAD-Fmk and JNK inhibition. Moreover, ruthenium red protected PMH against PA and MA-induced cell death. Recapitulating in vitro findings, mice fed a diet enriched in PA plus MA exhibited lipodystrophy, hepatosplenomegaly, increased liver ceramide content and cholesterol levels, ER stress, liver damage, inflammation and fibrosis compared to mice fed diets enriched in PA or MA alone. The deleterious effects of PA plus MA-enriched diet were largely prevented by in vivo myriocin treatment. These findings indicate a causal link between ceramide synthesis and ER stress in lipotoxicity, and imply that the consumption of diets enriched in MA and PA can cause associated with lipodystrophy.

Keyword: NASH

Nonalcoholic fatty liver disease impairs the cytochrome P-450-dependent metabolism of α-tocopherol (vitamin E).

This study aims to investigate in in vivo and in vitro models of nonalcoholic fatty liver disease (NAFLD) the enzymatic metabolism of α-tocopherol (vitamin E) and its relationship to vitamin E-responsive genes with key role in the lipid metabolism and detoxification of the liver. The experimental models included mice fed a high-fat diet combined or not with fructose (HFD+F) and HepG2 human hepatocarcinoma cells treated with the lipogenic agents palmitate, oleate or fructose. CYP4F2 protein, a cytochrome P-450 isoform with proposed α-tocopherol ω-hydroxylase activity, decreased in HFD and even more in HFD+F mice liver; this finding was associated with increased hepatic levels of α-tocopherol and decreased formation of the corresponding long-chain metabolites α-13-hydroxy and α-13-carboxy chromanols. A decreased expression was also observed for PPAR-γ and SREBP-1 proteins, two vitamin E-responsive genes with key role in lipid metabolism and CYP4F2 gene regulation. A transient activation of CYP4F2 gene followed by a repression response was observed in HepG2 cells during the exposure to increasing levels of the lipogenic and cytotoxic agent ; such gene repression effect was further exacerbated by the co-treatment with oleic and α-tocopherol and was also observed for PPAR-γ and the SREBP isoforms 1 and 2. Such gene response was associated with increased uptake and ω-hydroxylation of α-tocopherol, which suggests a minor role of CYP4F2 in the enzymatic metabolism of vitamin E in HepG2 cells. In conclusion, the liver metabolism and gene response of α-tocopherol are impaired in experimental NAFLD.Copyright © 2017. Published by Elsevier Inc.

Keyword: NASH

Role of fibroblast growth factor 21 in the early stage of induced by methionine- and choline-deficient diet.

Fibroblast growth factor 21 (FGF21) is a modulator of energy homeostasis and is increased in human nonalcoholic liver disease (NAFLD) and after feeding of methionine- and choline-deficient diet (MCD), a conventional inducer of murine nonalcoholic steatohepatitis (). However, the significance of FGF21 induction in the occurrence of MCD-induced remains undetermined. C57BL/6J Fgf21-null and wild-type mice were treated with MCD for 1 week. Hepatic Fgf21 mRNA was increased early after commencing MCD treatment independent of peroxisome proliferator-activated receptor (PPAR) α and farnesoid X receptor. While no significant differences in white adipose lipolysis were seen in both genotypes, hepatic triglyceride (TG) contents were increased in Fgf21-null mice, likely due to the up-regulation of genes encoding CD36 and phosphatidic phosphatase 2a/2c, involved in fatty (FA) uptake and diacylglycerol synthesis, respectively, and suppression of increased mRNAs encoding carnitine palmitoyl-CoA transferase 1α, PPARγ coactivator 1α, and adipose TG lipase, which are associated with lipid clearance in the liver. The MCD-treated Fgf21-null mice showed increased hepatic endoplasmic reticulum (ER) stress. Exposure of primary hepatocytes to elevated the mRNA levels encoding DNA damage-inducible transcript 3, an indicator of ER stress, and FGF21 in a PPARα-independent manner, suggesting that lipid-induced ER stress can enhance hepatic FGF21 expression. Collectively, FGF21 is elevated in the early stage of MCD-induced likely to minimize hepatic lipid accumulation and ensuing ER stress. These results provide a possible mechanism on how FGF21 is increased in NAFLD/.Published by Elsevier B.V.

Keyword: NASH

Hepatocyte nuclear receptor SHP suppresses inflammation and fibrosis in a mouse model of nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is a burgeoning health problem worldwide, ranging from nonalcoholic fatty liver (NAFL, steatosis without hepatocellular injury) to the more aggressive nonalcoholic steatohepatitis (, steatosis with ballooning, inflammation, or fibrosis). Although many studies have greatly contributed to the elucidation of NAFLD pathogenesis, the disease progression from NAFL to remains incompletely understood. Nuclear receptor small heterodimer partner (Nr0b2, ) is a transcriptional regulator critical for the regulation of bile , glucose, and lipid metabolism. Here, we show that SHP levels are decreased in the livers of patients with and in diet-induced mouse . Exposing primary mouse hepatocytes to and lipopolysaccharide , we demonstrated that the suppression of expression in hepatocytes is due to c-Jun N-terminal kinase (JNK) activation, which stimulates c-Jun-mediated transcriptional repression of Interestingly, induction of hepatocyte-specific SHP in steatotic mouse liver ameliorated progression by attenuating liver inflammation and fibrosis, but not steatosis. Moreover, a key mechanism linking the anti-inflammatory role of hepatocyte-specific SHP expression to inflammation involved SHP-induced suppression of NF-κB p65-mediated induction of chemokine (C-C motif) ligand 2 (CCL2), which activates macrophage proinflammatory polarization and migration. In summary, our results indicate that a JNK/SHP/NF-κB/CCL2 regulatory network controls communications between hepatocytes and macrophages and contributes to the disease progression from NAFL to . Our findings may benefit the development of new management or prevention strategies for .© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: NASH

Skeletal muscle miR-34a/SIRT1:AMPK axis is activated in experimental and human non-alcoholic steatohepatitis.

Non-alcoholic fatty liver disease (NAFLD) pathogenesis associates with intramyocellular lipid deposition and mitochondrial dysfunction. microRNAs (miRs), including pro-apoptotic miR-34a, are modulated during disease progression in liver tissue and plasma. We aimed to investigate the functional role of the miR-34a/SIRT1:AMP-activated protein kinase (AMPK) pathway in modulating local mitochondrial dysfunction in the skeletal muscle of human and experimental non-alcoholic steatohepatitis. Muscle biopsies were obtained from morbid obese NAFLD patients undergoing bariatric surgery. C57BL/6N mice were fed different NAFLD-inducing diets and C2C12 muscle cells incubated with (PA) in the presence or absence of an AMPK activator, or upon miR-34a functional modulation. Several muscle miRNAs, including miR-34a, were found increased with human NAFLD progression. Activation of the miR-34a/SIRT1:AMPK pathway, concomitant with impairment in insulin signalling mediators and deregulation of mitochondrial-shaping proteins, was evident in C2C12 cells incubated with PA, as well as in the skeletal muscle of all three diet-induced NAFLD mice models. Functional studies established the association between miR-34a- and PA-induced muscle cell deregulation. Of note, activation of AMPK almost completely prevented miR-34a- and PA-induced cellular stress. In addition, the miR-34a/SIRT1:AMPK pathway and mitochondrial dynamics dysfunction were also found amplified in muscle of human NAFLD. Finally, muscle miR-34a expression and mitofusin 2 (Mfn2) protein levels correlated with hallmarks of NAFLD and disease progression. Our results indicate that activation of the miR-34a/SIRT1:AMPK pathway leads to mitochondrial dynamics dysfunction in skeletal muscle of human and experimental NAFLD, representing an appealing prospective target in metabolic syndrome. KEY MESSAGES: Skeletal muscle microRNAs are modulated during NAFLD progression. -induced muscle cell dysfunction occurs, at least in part, through activation of the miR-34a/SIRT1:AMPK pathway. miR-34a/SIRT1:AMPK activation associates with mitochondria dynamics dysfunction in human NAFLD.

Keyword: NASH

Fat-loaded HepG2 spheroids exhibit enhanced protection from Pro-oxidant and cytokine induced damage.

The mechanisms by which steatosis renders hepatocytes susceptible to damage in non-alcoholic steatohepatitis () are unclear although fat accumulation is believed to increase hepatocyte susceptibility to inflammatory cytokines and oxidative stress. We therefore investigated the susceptibility of steatotic, hepatocyte-derived cells to TNFalpha and the pro-oxidant, t-butylhydroperoxide (TBH). HepG2 spheroids rendered steatotic by fat-loading with 0.15 mM oleic or for 48 h and treated with TNFalpha or TBH for 18 h exhibited surprisingly lower levels of cytotoxicity, and increased anti-oxidant activity (superoxide dismutase (SOD)) compared with non fat-loaded controls. The protective effect of steatosis was significantly reversed by the inhibition of AMP-activated kinase (AMPK) since spheroids transfected with a kinase-dead AMPKalpha2 subunit, exhibited a significant increase in TBH-induced cytotoxicity when fat-loaded. In conclusion, our findings suggest that fat-loaded hepatocyte-derived cells are surprisingly less susceptible to cytokine and pro-oxidant induced damage via an adaptive mechanism dependent, in part, on AMPK activity.

Keyword: NASH

Niacin inhibits fat accumulation, oxidative stress, and inflammatory cytokine IL-8 in cultured hepatocytes: Impact on non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a common disorder characterized by excessive hepatic fat accumulation, production of reactive oxygen species (ROS), inflammation and potentially resulting in non-alcoholic steatohepatitis (), cirrhosis and end-stage liver disease. Recently, we have shown that niacin significantly prevented hepatic steatosis and regressed pre-existing steatosis in high-fat fed rat model of NAFLD. To gain further insight into the cellular mechanisms, this study investigated the effect of niacin on human hepatocyte fat accumulation, ROS production, and inflammatory mediator IL-8 secretion.Human hepatoblastoma cell line HepG2 or human primary hepatocytes were first stimulated with followed by treatment with niacin or control for 24 h.The data indicated that niacin (at 0.25 and 0.5 mmol/L doses) significantly inhibited -induced fat accumulation in human hepatocytes by 45-62%. This effect was associated with inhibition of diacylglycerol acyltransferase 2 (DGAT2) mRNA expression without affecting the mRNA expression of fatty synthase (FAS) and carnitine palmitoyltransferase 1 (CPT1). Niacin attenuated hepatocyte ROS production and it also inhibited NADPH oxidase activity. Niacin reduced -induced IL-8 levels.These findings suggest that niacin, through inhibiting hepatocyte DGAT2 and NADPH oxidase activity, attenuates hepatic fat accumulation and ROS production respectively. Decreased ROS production, at least in part, may have contributed to the inhibition of pro-inflammatory IL-8 levels. These mechanistic studies may be useful for the clinical development of niacin and niacin-related compounds for the treatment of NAFLD/ and its complications.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: NASH

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving lipid metabolism, antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis () is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high fat high fructose (HFHF) diet fed C57BL/6J mice, used as model of . The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved lipid homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, mitochondria are intracellular source of reactive oxygen species (ROS) and important sites of lipid metabolism, we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti- potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: NASH

Hepatic FTO expression is increased in and its silencing attenuates -induced lipotoxicity.

Non-alcoholic steatohepatitis () is one of the most common causes of liver failure worldwide. It is characterized by excess fat accumulation, inflammation, and increased lipotoxicity in hepatocytes. Currently, there are limited treatment options for due to lack of understanding of its molecular etiology. In the present study, we demonstrate that the expression of fat mass and obesity associated gene (FTO) is significantly increased in the livers of patients and in a rodent model of . Furthermore, using human hepatic cells, we show that genetic silencing of FTO protects against palmitate-induced oxidative stress, mitochondrial dysfunction, ER stress, and apoptosis in\xa0vitro. Taken together, our results show that FTO may have a deleterious role in hepatic cells during lipotoxic conditions, and strongly suggest that up-regulation of FTO may contribute to the increased liver damage in .Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: NASH

Effects of deep-bedded finishing system on market pig performance, composition and pork quality.

The purpose of this study was to compare effects of finishing environment on growth performance, pork quality and lipid composition of pork. Environments compared were standard confinement (CON) and deep-bedded semi-outdoor systems. The deep-bedded method employed in the current study was the use of hoop structures. Hoops are large, tent-like shelters with cornstalks or straw for bedding. Gilts ranging in weight from 59 to 71 kg were randomly assigned to treatments of Hoop (n = 50) and CON (n = 18) environments. Gilts were fed a two-phase dietary sequence, ad libitum for 45 days. Six gilts per treatment were selected for carcass composition and quality evaluation. The experiment was replicated a total of five times. Pigs raised in the Hoop environment gained significantly less and required significantly more feed for growth than pigs raised in the CON environment. Carcasses from CON-finished pigs were significantly fatter at the 10th rib, which lowered carcass percentage fat-free lean(FFL) and they also had greater loin marbling scores compared with carcasses from Hoop-finished pigs (P < 0.05). Significant replication effects were noted on beginning weight, live weight, carcass weight, percentage FFL, backfat, lipid content and adipose firmness. Carcasses from Hoop pigs had lower proportions of (P < 0.05), and higher proportions of oleic and linoleic (P < 0.05) in the inner layer of adipose tissue. The proportion of saturated was lower, and that of mono- and poly-unsaturated was higher in the inner layer of the adipose tissue of Hoop pigs. Variations in composition and lipid deposition may have been caused by environmental temperature, since decreases in environmental temperature accompanied compositional variation of the adipose, leading to higher proportions of monounsaturated and lower proportions of saturated and polyunsaturated in adipose tissue, regardless of treatment. profile analysis revealed that adipose tissue of Hoop pigs had significantly higher amounts of 3-butanal and heptanal compared with CON pigs, which may be related to the amount of oleic and linoleic composing the adipose tissue. These data indicate finishing pigs in hoop structures allows for exposure to fluctuating temperatures, which may influence the growth of pigs, as well as composition and firmness of pork products.

Keyword: SCFA

Effects of milling on aromatics, lipophilic phytonutrients, and in unprocessed white rice of scented rice \'Cheonjihyang-1-se.

To understand effects of milling, scented rice \'Cheonjihyang-1-se\' was milled from 10 to 140\xa0s and changes in volatiles, phytonutrients, and were evaluated. Among 43 identified odor-active compounds, four volatiles, including hexan-3-one, exhibited decreases of up to 78%, while four others including ()-non-2-enal, increased following milling. Levels of 2-acetyl-1-pyrroline, the most distinctive popcorn-flavoring compound in scented rice, were not affected by the degree of milling (DM). Partial least squares discriminant analyses of volatiles were able to differentiate white rice according to the DM. Benzene and 2-pentylfuran showed the highest variable importance in projection scores, which could be applied in estimating the DM of rice. Milling significantly decreased tocopherols, tocotrienols, squalene, phytosterols contents and oleic composition, while composition was increased. These results suggest milling-dependent variations in phytonutrient levels and lipid composition, as well as changes in aroma and subsequent market quality, in \'Cheonjihyang-1-se\' rice.

Keyword: SCFA

and Oil Compositions of Allomyrina dichotoma Larvae.

Thirty-two different oils were identified from Allomyrina dichotoma (A. dichotoma) larvae by gas chromatography/mass spectrometry (GC/MS). The major components were 2,2,4-trimethyl-3-carboxyisopropyl pentanoic isobutyl ester (5.83%), phenol,2,6-bis(a,a-dimethyl ethyl)-4-(1-methyl-1-phenylethyl) (5.72%), heptacosane (5.49%) and phenol,2,4-bis(1-methyl-1-phenylethyl) (5.47%). The composition of the in A. dichotoma larvae was also determined by gas chromatography (GC) and fourteen constituents were identified. Oleic (19.13%) was the most abundant followed by (12.52%), palmitoleic (3.71%) and linoleic (2.08%) in 100 g of A. dichotoma larvae on a dry weight basis. The quantity of unsaturated (64.00%) were higher than that of saturated ones (36.00%). The predominant in A. dichotoma consist of monounsaturated (MUFA, 57.70%) such as oleic , myristoleic and palmitoleic , followed by saturated (36.00%) and polyunsaturated (PUFA, 6.50%). In particular, the presence of essential , such as linoleic (5.30%) and linolenic (0.40%) give A. dichotoma larvae considerable nutritional and functional value and it may be a useful source for food and/or industrial utilization.

Keyword: SCFA

Comparing the effect of homogenization and heat processing on the properties and in vitro digestion of milk from organic and conventional dairy herds.

We compared the effects of homogenization and heat processing on the chemical and in vitro digestion traits of milk from organic and conventional herds. Raw milk from organic (>50% of dry matter intake from pasture) and conventional (no access to pasture) farms were adjusted to commercial whole and nonfat milk fat standards, and processed with or without homogenization, and with high-temperature-short-time or UHT pasteurization. The milk then underwent in vitro gastrointestinal digestion. Comparison of milk from organic and conventional herds showed that the milks responded to processing in similar ways. General composition was the same among the whole milk samples and among the nonfat milk samples. Protein profiles were similar, with intact caseins and whey proteins predominant and only minor amounts of peptides. Whole milk samples from grazing cows contained higher levels of α-linolenic (C18:3), vaccenic (C18:1 trans), and conjugated linoleic , and lower levels of (C16:0) and stearic (C18:0) than samples from nongrazing cows. Processing had no effect on conjugated linoleic and linolenic levels in milk, although homogenization resulted in higher levels of C8 to C14 saturated . Of the 9 compounds evaluated, milk from grazing cows contained lower levels of 2-butanone than milk from nongrazing cows, and milk from both farms showed spikes for heptanal in UHT samples and spikes for butanoic, octanoic, nonanoic, and N-decanoic in homogenized samples. At the start of in vitro digestion, nonfat raw and pasteurized milk samples formed the largest clots, and organic milk clots were larger than conventional milk clots; UHT whole milk formed the smallest clots. Milk digests from grazing cows had lower levels of free than digests from nongrazing cows. In vitro proteolysis was similar in milk from both farms and resulted in 85 to 95% digestibility. Overall, milk from organic/grass-fed and conventional herds responded in similar ways to typical homogenization and heat processing used in United States dairy plants and showed only minor differences in chemical traits and in vitro digestion. Findings from this research enhance our knowledge of the effect of processing on the quality traits and digestibility of milk from organic/pasture-fed and confined conventional herds and will help health-conscious consumers make informed decisions about dairy selections.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Nutritional evaluation of processing discards from tiger tooth croaker, .

Processing discards from tiger tooth croaker, such as head, viscera, and swim bladder, were analyzed for proximate, biochemical, , and mineral composition. The proximate analysis showed high protein content (21.04%) in the swim bladder and high fat content (4.10%) and ash content (2.26%) in the head. The biochemical quality indices such as non-protein nitrogen, total base, trimethylamine, free , peroxide value, and thiobarbituric reactive substances in the viscera were higher than those in the head and swim bladder. Gas chromatographymass spectrometry analysis of composition showed high polyunsaturated in the head (58.82%), followed by the viscera (45.80%) and the swim bladder (35.57%). The major available saturated , i.e., , was higher in the swim bladder (30.49%). Mineral and heavy metals analyzed via inductively coupled plasma-optical emission spectrometer showed high calcium, sodium, and magnesium contents in the head region (346.80, 62.55, and 14.89 mg/kg respectively). Heavy metal levels were within the permissible limit.

Keyword: SCFA

Evaluation of Profile, Composition and in vitro Bioactivity of Growing Wild in Northern Iran.

The aim of the present study was to investigate the chemical properties of wild Tagetes minuta L. (family Astreacea) collected from Northern Iran during the flowering period concerning the chemical combination of the essential oil along with its antioxidant properties and composition of . The essential oil of the plant was extracted by a Clevenger approach and analyzed using gas chromatography-mass spectroscopy (Capillary HP-5ms GC/MS Column). contents of this species as a result of hexane extraction were analyzed by means of gas chromatography (GC-FID) while their phenolic contents were analyzed by high performance liquid chromatography (HPLC-UV). In this research also the total polyphenolic (TPC) and total flavonoid (TFC) content was determined spectrophotometrically while the antioxidant activity was evaluated using the DPPH (2,2\'-diphenyl-1-picrylhydrazyl) bleaching method. GC/MS analysis of the essential oil identified monoterpenoid fractions (52.13%) as the main components and among them dihydrotagetone (23.44%) and spathulenol (10.56%) were the predominant compounds. The evaluation of content revealed that saturated were prevailing compounds and the major components are: (30.74±0.4%) and capric (24.15±0.5%) . Chromatographic separation of its phenolic contents indicated that this herb contain sinapic derivatives rather than hydroxybenzoic derivatives. Also the essential oil showed an effective antioxidant capacity (TPC=153.27±0.9 mg/g, TFC=63.79±0.1 mg/g, IC = 29.31±0.8 µg/ml). The results proved that the plant could be used for nutritional and pharmaceutical purposes.

Keyword: SCFA

The impact of dietary sn-2 triacylglycerols in combination with docosahexaenoic or arachidonic on lipid metabolism and host faecal composition in Sprague Dawley rats.

Sn-2 triacylglycerols (sn2PA fat) and polyunsaturated fatty acids are thought to influence the metabolic status and intestinal bacterial population of the host. In this study, the impact of sn2PA fat in combination with DHA or ARA in the diet on lipid metabolism in the liver and faecal composition were investigated in rats fed diets containing sn2PA fat, 90% sn2PA fat + 10% DHA oil (wt%), or 90% sn2PA fat + 10% ARA oil (wt%). Tissue fatty composition was measured using gas chromatography (GC), whereas the faecal microbial composition was assessed using 16S rRNA high-throughput sequencing technology. In addition, faecal short-chain fatty acids (SCFA) were analyzed using ion chromatography. The results showed that sn2PA fat in combination with DHA or ARA significantly reduced liver triacylglyceride (TG) content compared with the sn2PA fat only group. Moreover, the supplementation with sn2PA fat in combination with DHA or ARA significantly promoted the growth of Lactobacillus in the feces at the genus level. On the other hand, the growth of the opportunistic pathogen Desulfovibrio was significantly inhibited by sn2PA fat in combination with ARA compared with the sn2PA fat group. In addition, sn2PA fat in combination with DHA or ARA significantly increased total SCFA concentration in the faeces, suggesting a beneficial effect on host intestinal health.

Keyword: SCFA

Ingested medium-chain are directly utilized for the acyl modification of ghrelin.

Ghrelin, an acylated brain and gut peptide, is primarily produced by endocrine cells of the gastric mucosa for secretion into the circulation. The major active form of ghrelin is a 28-amino- peptide containing an n-octanoyl modification at serine that is essential for activity. Studies have identified multiple physiological functions for ghrelin, including GH release, appetite stimulation, and metabolic fuel preference. Until now, there has not been any report detailing the mechanism of ghrelin acyl modification. Here we report that ingestion of either medium-chain (MCFAs) or medium-chain triacylglycerols (MCTs) increased the stomach concentrations of acylated ghrelin without changing the total (acyl- and des-acyl-) ghrelin amounts. After ingestion of either MCFAs or MCTs, the carbon chain lengths of the acyl groups attached to nascent ghrelin molecules corresponded to that of the ingested MCFAs or MCTs. Ghrelin peptides modified with n-butyryl or n-palmitoyl groups, however, could not be detected after ingestion of the corresponding short-chain or long-chain , respectively. Moreover, n-heptanoyl ghrelin, an unnatural form of ghrelin, could be detected in the stomach of mice after ingestion of either n-heptanoic or glyceryl triheptanoate. These findings indicate that ingested medium-chain are directly used for the acylation of ghrelin.

Keyword: SCFA

Lipid metabolism in response to individual short chain during mixotrophic mode of microalgal cultivation: Influence on biodiesel saturation and protein profile.

Critical influence of different short chain as organic carbon source, during growth (GP) and nutrient stress lipogenic phase (NSLP) was investigated on biomass and lipid productivity, in mixotrophic fed-batch microalgae cultivation. Nutrient deprivation induced physiological stress stimulated highest lipid productivity with acetate (total/neutral lipids, 35/17) with saturation index of 80.53% by the end of NSLP followed by butyrate (12/7%; 78%). Biomass growth followed the order of acetate (2.23 g/l) >butyrate (0.99 g/l) >propionate (0.77 g/l). VFA removal (as COD) was maximum with acetate (87%) followed by butyrate (55.09%) and propionate (10.60%). was the most dominant found in the composition of all variants and butyrate fed system yielded a maximum of 44% . Protein profiling illustrated prominence of acetyl CoA-synthetase activity in acetate system. Thus, provide a promising alternative feedstock for biodiesel production with integrated microalgae-biorefinery.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Bioformation of and Nonvolatile Metabolites by KJJ81 Cultivated under Different Conditions-Carbon Sources and Cultivation Times.

KJJ81 isolated from is an amylolytic yeast that is widely used as a microbial starter in various fermented foods. and nonvolatile metabolites of KJJ81 were investigated according to different carbon sources and cultivation times using a nontargeted metabolomic approach. Partial-least-squares discriminant analysis was applied to determine the major metabolites, which were found to be closely related to the clustering and discrimination of KJJ81 samples. Some metabolites derived from phenylalanine, such as 2-phenylethanol, 2-phenylethyl acetate, and ethyl phenylacetate, were predominantly found in cultivation medium containing glucose (YPD medium). In addition, the level of 2-phenylethanol increased continuously with the cultivation time. In terms of nonvolatile metabolites, carbohydrates (mannose, arabitol, and mannitol), ( and stearic ), organic (oxalic and succinic ), and amino (isoleucine, serine, alanine, glutamic , glycine, proline, phenylalanine, and threonine) were the main contributors to KJJ81 samples cultivated in YPD medium according to cultivation time. These results show that the formation of and nonvolatile metabolites of KJJ81 can be significantly affected by both the carbon sources and the cultivation time.

Keyword: SCFA

Dependence of arbuscular-mycorrhizal fungi on their plant host for synthesis.

Lipids are the major form of carbon storage in arbuscular-mycorrhizal fungi. We studied synthesis by Glomus intraradices and Gigaspora rosea. [(14)C]Acetate and [(14)C]sucrose were incorporated into a synthetic culture medium to test synthetic ability in germinating spores (G. intraradices and G. rosea), mycorrhized carrot roots, and extraradical fungal mycelium (G. intraradices). Germinating spores and extraradical hyphae could not synthesize 16-carbon but could elongate and desaturate already present. The growth stimulation of germinating spores by root exudates did not stimulate synthesis. 16-Carbon (16:0 and 16:1) were synthesized only by the fungi in the mycorrhized roots. Our data strongly suggest that the synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that metabolism may play a major role in the obligate biotrophism of arbuscular-mycorrhizal fungi.

Keyword: SCFA

Ethanol shock changes the profile and survival behavior of Vibrio parahaemolyticus in various stress conditions.

Vibrio parahaemolyticus 690, a clinical strain, was subjected to ethanol shock in the presence of 5% ethanol for a period of 30 and 60 min. Survival behaviors of the ethanol shocked and control cells of V. parahaemolyticus in the presence of H(2)O(2) (20 ppm), crystal violet (3 ppm), NaCl (20%), and low pH solution (pH 4.4) containing various organic including lactic , acetic , citric and tartaric (25 mM) were compared. In addition, the effects of ethanol shock on the profile and recovery of V. parahaemolyticus on tryptic soy agar (TSA) plus various amounts of NaCl were also investigated. After ethanol shock, it was found that the proportion of vaccenic (18:1) increased, while the proportion of (16:0) and ratio of saturated to unsaturated decreased in cells of V. parahaemolyticus. The recovery of the ethanol-shocked cells on TSA plus 6.0% or 7.5% NaCl was significantly less than the control cells. Furthermore, ethanol shock increased the survival of V. parahaemolyticus in the presence of H(2)O(2), while made the test organism less resistant to crystal violet, high NaCl and organic . The degree of decreased tolerance observed on the ethanol-shocked cells of test organism varied with the organic examined. Finally, ethanol shock for 60 min exhibited either a higher or similar degree of effect on the test organism (depending on the type of stress encountered) than did ethanol shock for 30 min. Data obtained from the present study does provide useful information that is indispensable when control measure of V. parahaemolyticus is to be performed efficiently and adequately.

Keyword: SCFA

Bioconversion of derived from waste activated sludge into lipids by Cryptococcus curvatus.

Pure (VFA) solution derived from waste activated sludge (WAS) was used to produce microbial lipids as culture medium in this study, which aimed to realize the resource recovery of WAS and provide low-cost feedstock for biodiesel production simultaneously. Cryptococcus curvatus was selected among three oleaginous yeast to produce lipids with VFAs derived from WAS. In batch cultivation, lipid contents increased from 10.2% to 16.8% when carbon to nitrogen ratio increased from about 3.5 to 165 after removal of ammonia nitrogen by struvite precipitation. The lipid content further increased to 39.6% and the biomass increased from 1.56g/L to 4.53g/L after cultivation for five cycles using sequencing batch culture (SBC) strategy. The lipids produced from WAS-derived VFA solution contained nearly 50% of monounsaturated , including , heptadecanoic , ginkgolic , stearic , oleic , and linoleic , which showed the adequacy of biodiesel production.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Seasonal variation of the chemical composition and antimicrobial and cytotoxic activities of the essential oils from Inga laurina (Sw.) Willd.

The seasonal chemical composition of essential oils from Inga laurina was determined by GC/MS. In the stem bark\'s essential oil extracted during the dry season, the presence of terpenoids (30.05%) stood out, and phytol (9.76%) was the major compound identified. For the stem bark oil obtained during the rainy season, in addition to terpenoids (26.63%), a large amount of (46.84%) were identified, in particular (25.40%). Regarding the leaves\' essential oil obtained in the dry season, esters (42.35%) were the main components. The main ester present was (Z)-hex-3-enyl benzoate (10.15%) and the major compound of this oil was (Z)-hex-3-en-1-ol (14.23%). Terpenoids (33.84%), long-chain alkanes (27.04%) and (21.72%) were the main components of the essential oil from leaves in the rainy season. Phytol (33.21%), nonacosane (21.95%) and (15.20%) were the major compounds identified. The antimicrobial activity against aerobic and anaerobic oral bacteria was evaluated by the microdilution broth method and cytotoxic activity was carried out with Vero cells. The essential oils from the rainy season showed a better inhibition of the bacterial growth with Minimal Inhibitory Concentrations (MIC) values of 25 or 50 µg·mL⁻¹ for aerobic bacteria, and high selectivity against bacteria was observed. The large amount of in rainy season oils may be related to the better inhibitory effects observed.

Keyword: SCFA

Chemical composition of volatiles from Opuntia littoralis, Opuntia ficus-indica, and Opuntia prolifera growing on Catalina Island, California.

The essential oils from the cladodes of Opuntia littoralis, Opuntia ficus-indica and Opuntia prolifera growing wild on Santa Catalina Island, California, were obtained by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC-MS). Terpenoids were the dominant class of volatiles in O. littoralis, with the two main components being the furanoid forms of cis-linalool oxide (10.8%) and trans-linalool oxide (8.8%). -derived compounds dominated the essential oil of O. ficus-indica with linoleic (22.3%), (12.7%), lauric (10.5%) and myristic (4.2%) as major . O. prolifera oil was composed of 46.6% alkanes and the primary hydrocarbon component was heptadecane (19.2%). Sixteen compounds were common to all the three Opuntia species.

Keyword: SCFA

Synthesis of chitosan-based polymeric surfactants and their adsorption properties for heavy metals and .

Chitosan-based polymeric surfactants (CBPSs) were prepared by N-acylation of chitosans (chitosan 10 and 500) with several anhydrides such as hexanoic (C6), lauric (C12), and (C16) anhydrides. Among the CBPS samples, CBPSs having a good solubility at pH 4.0 were selected and observed for viscosity, surface tension, and adsorption of heavy metals (Cd2+, Co2+, Cr2O7(2-), and Pb2+) as well as the (n-octanoic ). The 1H NMR spectrum of chitosan 10 modified with C16 at the substitution ratio of 0.4 (CBPS10-C16,0.4) showed 85% of acylation in 1% DCl/D2O solutions. CBPS10 with the substitution ratio less than 0.4 showed a good solubility because of shorter repeating units and lesser amounts of hydrophobic substituents. The intrinsic viscosity of CBPS10 was slightly increased, while that of CBPS500 was decreased. As the substitution ratio and length of the carbon chain increased, the surface tension of CBPS10 tended to decrease. CBPS10-C16,0.2 had high adsorption ability for cationic metal ions such as Cd2+, Co2+, and Pb2+ comparable to chitosan. Interestingly, CBPS(10)-C(16,0.2) showed a unique pH optimum for the anionic metal ion such as Cr2O7(2-). In addition, CBPS10-C16,0.2 exhibited the highest adsorption ability for n-octanoic among the tested CBPS10 with different carbon chains.

Keyword: SCFA

[Chemical constituents of Eupatorium lindleyanum].

To study chemical constituents of Eupatorium lindleyanum. Ethyl acetate extractive fractions were separated with silica gel and Sephadex LH-20 by column chromatography, and their structures were identified on the basis of spectroscopic analysis and chemical evidence. Sixteen compounds were separated and identified as scopoletin (1), 6, 7-dimethylesculetin (2), nepetin (3), eupatrin (4), luteolin (5), isoquerecitrin (6), jaceosidin (7), quceritin (8), kaempferol (9), rutin (10), cirsiliol (11), taraxasterylacetate (12), pseudotaraxasteryl acetate (13), pseudotaraxasterol (14), butanoic (15) and n-hexadecanoic (16). Of them, compounds 1-6 and 11, 13 and 15 were separated from this plant for the first time.

Keyword: SCFA

Metabolite profiling of doenjang, fermented soybean paste, during fermentation.

A fermented soybean paste known as doenjang is a traditional fermented food that is widely consumed in Korea. The quality of doenjang varies considerably by its basic ingredients, species of microflora, and fermentation process. The classification of predefined metabolites (e.g. amino , organic , sugars and sugar derivatives, and ) in doenjang samples according to fermentation was performed by using GC-FID and GC-MS data sets with the application of a multivariate statistical method.The predominantly produced amino included alanine, valine, leucine, isoleucine, proline, glutamine, phenylalanine and lysine, showing remarkable increases in amounts during the later stages of fermentation. Carbonic , citric , lactic and pyrogultamic were identified as the major organic . Significant amounts of erythrose, xylitol, inositol and mannitol were detected during fermentation. Regarding , relatively higher amounts of , stearic , oleic , linoleic and linolenic were found in the doenjang at each fermentation time point. Principal component analysis (PCA) successfully demonstrated changes in composition patterns as well as differences in non- metabolites according to fermentation period.A set of metabolites could be determined representing the quality of doenjang during fermentation, and which might also be correlated with taste ingredients, flavour, nutrition, and physiology activities that are claimed to be dependent on the quality control of commercial doenjang.Copyright (c) 2010 Society of Chemical Industry.

Keyword: SCFA

GC-MS Profiling of Volatile Components in Different Fermentation Products of Cordyceps Sinensis Mycelia.

The fermentation products of () mycelia are sustainable substitutes for natural . However, the volatile compositions of the commercial products are still unclear. In this paper, we have developed a simultaneous distillation-extraction (SDE) and gas chromatography-mass spectrometry (GC-MS) method for the profiling of volatile components in five fermentation products. A total of 64, 39, 56, 52, and 44 components were identified in the essential oils of Jinshuibao capsule (JSBC), Bailing capsule (BLC), Zhiling capsule (ZLC), Ningxinbao capsule (NXBC), and Xinganbao capsule (XGBC), respectively. 5,6-Dihydro-6-pentyl-2H-pyran-2-one (massoia lactone) was first discovered as the dominant component in JSBC volatiles. Fatty acids including (C16:0) and linoleic (C18:2) were also found to be major volatile compositions of the fermentation products. The multivariate partial least squares-discriminant analysis (PLS-DA) showed a clear discrimination among the different commercial products as well as the counterfeits. This study may provide further chemical evidences for the quality evaluation of the fermentation products of mycelia.

Keyword: SCFA

Evaluation of a screening method by liquid chromatography-tandem mass spectrometry for estimating effect of drugs on the activation and β-oxidation of in mitochondria.

metabolism is controlled not only by the acyl-coenzyme A (CoA) synthetases but by some enzymes in the β-oxidation cycle. Medium-chain and long-chain acyl-CoA esters are key metabolites in metabolism. We have developed an enzymatic assay method for determining chain shortening of the acyl-CoAs via β-oxidation from and octanoic in liver mitochondria. We have evaluated the assay method for detecting whether drugs influence the activation or the β-oxidation of .Liver mitochondria were used for investigating the effect of drugs on metabolism. The drugs selected were salicylic , diclofenac, valproic and paracetamol. Each acyl-CoA formed was analysed by liquid chromatography-tandem mass spectrometry.After less than 5 min of incubation, the levels of acyl-CoAs reflected the acyl-CoA synthetase activity, whereas after 60-min incubation they reflected the activity of some enzymes in the β-oxidation cycle. Salicylic , diclofenac and valproic inhibited the medium-chain acyl-CoA synthetases, whereas valproic only exhibited a weak inhibitory activity toward the β-oxidation of the medium-chain . In the case of long-chain metabolism, salicylic and diclofenac inhibited both the activation and β-oxidation, whereas valproic was a weak inhibitor for only the β-oxidation activity. Paracetamol showed hardly any influence on the metabolism of medium-chain and long-chain .These findings suggest that salicylic , diclofenac, valproic and paracetamol exert a different influence on metabolism depending on the length of the acyl chain. This assay allows sensitive and selective analysis for predicting the pathways by which drugs exert a greater influence over metabolism.© 2010 The Authors. JPP © 2010 Royal Pharmaceutical Society of Great Britain.

Keyword: SCFA

Tween 80 and respiratory growth affect metabolite production and membrane in Lactobacillus casei N87.

To evaluate the effect of cultivation (anaerobiosis vs respiration) and Tween 80 supplementation on the production of metabolites and on the composition of membrane (FAs) in Lactobacillus casei N87.Anaerobic and respiratory growth, with or without Tween 80 supplementation, was carried out in a chemically defined medium. Production of biomass, organic , organic compounds (VOCs), consumption of amino and changes in membrane FAs were investigated. Respiration altered the central metabolism rerouting pyruvate away from lactate accumulation, while Tween 80 had a minor effect on metabolic pathways. VOCs were mainly affected by growth conditions and significant amounts of diacetyl were produced by respiratory cultures. Respiration increased desaturation of membrane lipids and Tween 80 improved the production of essential polyunsaturated FAs. decreased in Tween-supplemented aerated cultures.Combination of Tween 80 and respiratory growth promoted production of biomass and aroma compounds and affected the composition of membrane FAs in Lact. casei N87.Respiration might be exploited in Lact. casei as a natural strategy for the enhanced production of aroma compounds.© 2016 The Society for Applied Microbiology.

Keyword: SCFA

Physico-chemical characterisation of the fat from red-skin rambutan (Nephellium lappaceum L.) seed.

The seeds (6.9±0.2% by weight of fruit) of the red-skin rambutan (Nephelium lappaceum L.) contain a considerable amount of crude fat (38.0±4.36%) and thus, the aim of the study was to determine the physico-chemical properties of this fat for potential applications. The iodine and saponification values, and unsaponifiable matter and free contents of the seed fat were 50.27 g I2/100g fat, 182.1 mg KOH/g fat, 0.8% and 2.1%, respectively. The fat is pale yellow with a Lovibond color index of 3.1Y+1.1R. The profile indicates an almost equal proportion of saturated (49.1%) and unsaturated (50.9%) , where oleic (42.0%) and arachidic (34.3%) were the most dominant . It also contained small amounts of stearic (8.0%), (4.6%), gadoleic (5.9%), linoleic (2.2%), behenic (2.1%) palmitoleic (0.7%) myristic (0.1%) and erucic (0.1%) . HPLC analysis showed that the fat comprised mainly unknown triacylglycerols (TAG) with high retention times indicating they have higher carbon numbers compared with many vegetable oils. The fat has melting and cooling points of 44.2°C and -42.5°C, respectively, making it a semi-solid at room temperature. The solid content at 0°C was 53.5% and the fat melted completely at 40°C. z-Nose analysis showed that the presence of high levels of compounds in red-skin rambutan seed and seed fat.

Keyword: SCFA

Changes in biochemical compounds in flesh and peel from Prunus persica fruits grown in Tunisia during two maturation stages.

Plants can synthesize tens to hundreds of thousands of primary and secondary metabolites with diverse biological properties and functions. (FA), phenolic compounds (PC) and compounds (VC) of flesh and peel from three Prunus persica cultivars were evaluated at the Regional Centre of Agricultural Research--Experimental Farm (Sidi Bouzid, Tunisia) during two maturation stages. , oleic and linoleic are the most abundant FA in Prunus persica cultivars. A genetic effect on FA composition was observed throughout the two sampling periods. Peel was rich in oleic with the highest content (31.3% on total FA) in \'O\'Henry\' cultivar at the commercial ripening date; flesh was rich in linoleic with the highest content (44.7% on total FA) in \'Sweet Cap\' cultivar at the full ripening date. The monounsaturated/polyunsaturated ratios were higher in the commercial ripe than in the full ripe fruits. The analysis of the composition of the VC led to the characterization of 98 different compounds, showing a very high variability among the cultivars. The full ripe fruit (peel and flesh) exhibited the highest total number of terpenoids. Commercial ripe peels were richest in the percentage of hydrocarbons. Comparing cultivars, \'Sweet Cap\' cultivar showed the lowest contents of alcohols in peel and flesh of full ripe fruit but highest in peel of commercial ripe fruit, and lowest content of aldehydes in peel and flesh of commercial ripe fruit but highest in peel of ripe ones and the highest ones of lactones. Among PC, the highest contents were observed for o-diphenols and the values showed varietal influence. Total phenols contents decreased during ripening process (p < 0.05) in both peel and flesh tissues, except found for \'Sweet Cap\' cultivar. In conclusion, to achieve better FA composition and greater VC and PC production of the peach fruit, P. persica cultivars should be harvested at the commercial ripening date.Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Keyword: SCFA

Substrate utilization and kinetics of surfactant metabolism in evolving bronchopulmonary dysplasia.

To use stable isotopically labeled precursors of pulmonary surfactant phospholipids to measure precursor utilization and surfactant turnover in premature infants who required mechanical ventilation at birth, 2 weeks, and >4 weeks of age.Infants of < or =28 weeks\' gestation received simultaneous 24-hour intravenous infusions of [1,2,3,4-13C4] palmitate and [1-13C1] acetate at birth, 2 weeks, and > or =4 weeks of life. Disaturated phospholipids were extracted from sequential tracheal aspirate samples obtained over a period of 2 weeks. Fractional catabolic rate (a measure of total turnover) and the fractional synthetic rates from plasma palmitate and de novo synthesis (acetate) were measured.The fractional catabolic rate increased from 25.3% +/- 7.0% per day at birth to 53.8% +/- 14.4% per day at 4 weeks (P=.001). The combined contribution from plasma palmitate and de novo synthesis to total synthesis increased from 44.2% +/- 19.8% at birth to 85.2% +/- 32.8% at 4 weeks (P=.03).Total surfactant turnover increased in premature infants with evolving bronchopulmonary dysplasia. The increasing contributions from acetate and plasma palmitate suggest a decrease in surfactant phospholipid recycling.

Keyword: SCFA

Roles and regulation of ketogenesis in cultured astroglia and neurons under hypoxia and hypoglycemia.

Exogenous ketone bodies (KBs), acetoacetate (AA), and β-hydroxybutyrate (BHB) act as alternative energy substrates in neural cells under starvation. The present study examined the endogenous ketogenic capacity of astroglia under hypoxia with/without glucose and the possible roles of KBs in neuronal energy metabolism. Cultured neurons and astroglia were prepared from Sprague-Dawley rats. (PAL) and l-carnitine (LC) were added to the assay medium. The 4- to 24-hr production of AA and BHB was measured using the cyclic thio-NADH method. (14)C-labeled -soluble products (KBs) and (14)CO2 produced from [1-(14)C]PAL were also measured. l-[U-(14)C]lactic ([(14)C]LAC), [1-(14)C]pyruvic ([(14)C]PYR), or β-[1-(14)C]hydroxybutyric ([(14)C]BHB) was used to compare the oxidative metabolism of the glycolysis end products with that of the KBs. Some cells were placed in a hypoxic chamber (1% O2). PAL and LC induced a higher production of KBs in astroglia than in neurons, while the CO2 production from PAL was less than 5% of the KB production in both astroglia and neurons. KB production in astroglia was augmented by the AMP-activated protein kinase activators, AICAR and metformin, as well as hypoxia with/without glucose. Neuronal KB production increased under hypoxia in the absence of PAL and LC. In neurons, [(14)C]LAC and [(14)C]PYR oxidation decreased after 24\u2009hr of hypoxia, while [(14)C]BHB oxidation was preserved. Astroglia responds to ischemia in\xa0vitro by enhancing KB production, and astroglia-produced KBs derived from might serve as a neuronal energy substrate for the tricarboxylic cycle instead of lactate, as pyruvate dehydrogenase is susceptible to ischemia.© The Author(s) 2014 Reprints and permissions: sagepub.com/journalsPermissions.nav.

Keyword: SCFA

Downregulation of matrix metalloproteinase-13 by the root extract of Cyathula officinalis Kuan and its constituents in IL-1β-treated chondrocytes.

The roots of Cyathula officinalis Kuan are widely used in Chinese medicine for the treatment of inflammatory disorders. Here, the ability of C. officinalis Kuan to downregulate matrix metalloproteinase (MMP)-13 was examined since MMP-13 is an important enzyme for the degradation of the cartilage collagen matrix, especially under arthritic conditions. The ethanol extract of C. officinalis Kuan as well as the N-hexane and chloroform soluble fractions were found to potently inhibit MMP-13 induction in IL-1 β-treated SW1353 cells, a human chondrosarcoma cell line, at 50-200\u2009µg/mL. Activity-guided separation led to the isolation of six compounds, (1), β-sitosterol (2), α-spinasterol (3), atractylenolide I (4), 1,3-diacetoxy-tetradeca-6E,12E-dien-8,10-dyn (5), and N-trans-feruloyl-3-methyldopamine (6). Among these, 4 and 5 exhibited MMP-13 downregulating activity in IL-1 β-treated SW1353 cells. And 4 also showed anti-oedematous activity against λ-carageenan-induced paw edema in mice at 20-200\u2009mg/kg, p.\u200ao. The results of this study provide information that can help elucidate the action mechanism of C. officinalis Kuan. In addition, the results presented here suggest that C. officinalis Kuan and its constituents may have the potential for chondroprotection against cartilage degrading disorders.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: SCFA

Short chain saturated decrease circulating cholesterol and increase tissue PUFA content in the rat.

This study investigates the effect of various dietary saturated (SFA) profiles on plasma lipid parameters and tissue composition in rats. The experiment was designed to monitor polyunsaturated (PUFA) levels, while examining different amounts and types of SFA. Four isocaloric diets were prepared, containing 10-11\xa0mol% of (FA) as linoleic (LNA) and 2.5\xa0mol% as α-linolenic (ALA), leading to an identical and well-balanced LNA/ALA ratio. The initial rapeseed oil/corn oil mixture providing ALA and LNA was enriched with olive oil to prepare the olive oil diet. The butterfat diet was supplemented with butterfat, containing short-chain SFA (C4:0-C10:0, 17\xa0mol% of FA), lauric (C12:0, 3.2\xa0mol%), myristic (C14:0, 10.5\xa0mol%) and (C16:0, 14.5\xa0mol%). The saturates diet was supplemented with trilaurin, trimyristin and tripalmitin to obtain the same level of lauric, myristic and as the butterfat diet, without the short-chain SFA. The trimyristin diet was enriched with trimyristin only. The results showed that the butterfat diet contributed to specific effects, compared to the olive oil diet and the saturates and trimyristin diets: a decrease in plasma total, LDL- and HDL-cholesterol, higher tissue storage of ALA and LNA, and a higher level of (n-3) highly unsaturated in some tissues. This study supports the hypothesis that in diets with identical well-balanced LNA/ALA ratios, short chain SFA may decrease circulating cholesterol and increase tissue polyunsaturated content in the rat.

Keyword: SCFA

Evidence for the involvement of GPR40 and NADPH oxidase in -induced superoxide production and insulin secretion.

G protein coupled receptor 40 (GPR40) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex have been shown to be involved in the amplification of glucose-stimulated insulin secretion (GSIS). The effect of on superoxide production and insulin secretion by INS-1E cells and the possible involvement of GPR40 and NADPH oxidase in these processes were examined in this study. Cells were incubated during 1 h with in low and high glucose concentrations, a GPR40 agonist (GW9508) and inhibitors of NADPH oxidase (diphenyleneiodonium, DPI) and PKC (calphostin C). GW9508 induced superoxide production at 2.8 and 5.6 mM glucose concentrations and stimulated insulin secretion at 16.7 mM glucose concentration involving both PKC and NADPH oxidase activation. induced superoxide production through NADPH oxidase and GPR40-dependent pathways and the stimulation of insulin secretion in the presence of a high glucose concentration was reduced by knockdown of GPR40 using siRNA. Our results suggest that induces superoxide production and potentiates GSIS through NADPH oxidase and GPR40 pathways in pancreatic ? cells.

Keyword: SCFA

Identification of Bioactivity, and Profile in Supercritical Fluid Extracts of Mexican arnica.

Supercritical fluid extraction (SFE) is a sustainable technique used for the extraction of lipophilic metabolites such as pigments and . Arnica plant is considered a potential candidate material with high antioxidant and antimicrobial activities. Therefore, in this study, a locally available Heterotheca inuloides, also known as Mexican arnica, was analyzed for the extraction of high-value compounds. Based on different pressure (P), temperature (T), and co-solvent (CoS), four treatments (T) were prepared. A maximum 7.13% yield was recovered from T2 (T = 60 °C, P = 10 MPa, CoS = 8 g/min), followed by 6.69% from T4 (T = 60 °C, P = 30 MPa, CoS = 4 g/min). Some bioactive sesquiterpenoids such as 7-hydroxycadalene, caryophyllene and δ-cadinene were identified in the extracts by GC/MS. The profile revealed that the main components were (C16:0), followed by linoleic (C18:2ω6c), α-linolenic (C18:3ω3) and stearic (C18:0) differing in percent yield per treatment. Antibacterial activities were determined by the agar diffusion method, indicating that all the treatments exerted strong antibacterial activity against S. aureus, C. albicans, and E. coli strains. The antioxidant capacity of the extracts was also measured by three in vitro assays, DPPH, TEAC and FRAP, using Trolox as a standard. Results showed high antioxidant capacity enabling pharmaceutical applications of Mexican arnica.

Keyword: SCFA

[Analysis of chemical constituents of the oil from flowers of Oyster Rhoeo by GC/MS].

To analyze the chemical constituents of the oil from the flowers of Oyster Rhoeo.The oil from the flower of Oyster Rhoeo was extracted by steam distillation and its chemical constituents were analyzed by GC/MS technique.All 20 compounds which relative content was greater than 0.3% were identified, accounting for 90.24% of the oil.The major constituent of the oil is nonaldehyde (25.72%), the other main constituents are hexadecanoid (22.79%), nonanoic (18.94%), dodecanoic (7.72%), isovanillin (2.84%), heptanoic (2.20%), octanoic (1.89%) and linoleic (1.71%), respectively.

Keyword: SCFA

Chemical composition of the essential oil and hexane extract of Salvia chionantha and their antioxidant and anticholinesterase activities.

The essential oil and methyl ester of hexane extract of Salvia chionantha Boiss. were analysed by GC and GC-MS. Totally, 54 components were detected in the essential oil and all of them were fully determined. Germacrene D (25.03%), β-caryophyllene (8.71%), spathulenol (5.86%) and α-humulene (4.82%) were identified as the major compounds. In the methylated hexane extract, 3-hydroxy hexadecanoic (39.39%), 3-hydroxy tetradecanoic (12.66%) and (12.02%) were the major elucidated. The antioxidant activity of the essential oil and the hexane extract was determined by using four complementary test systems; namely, β-carotene-linoleic , DPPH() scavenging, ABTS(+)* scavenging, and CUPRAC assays. In β-carotene-linoleic assay, the extract showed 81.2±0.1% lipid peroxidation inhibition at 0.8 mg/mL concentration, while in ABTS(+)* assay the essential oil exhibited 77.4±0.5% inhibition at same concentration. Since, acetylcholinesterase and butyrylcholinesterase enzymes are taking place in pathogenesis of Alzheimer\'s disease, in vitro anticholinesterase activity of the essential oil and the extract was also studied spectrophotometrically. At 0.5mg/mL concentration, the essential oil showed moderate acetylcholinesterase (56.7±1.9%) and butyrylcholinesterase (41.7±2.9%) inhibitory activity, while the extract was only exhibited activity (63.1±0.8%) against butyrylcholinesterase enzyme. Hence, the essential oil may be useful as a moderate anticholinesterase agent, particularly against acetylcholinesterase.Copyright © 2010 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Estrogen-like activity of components from Vitex rotundifolia L.

The fruits of Vitex rotundifolia L. are used as a traditional medicine in China. Dried fruits of V. rotundifolia (rich in essential oils) have been known to show strong estrogenic activity. Therefore, we investigated components in the essential oil isolated from the fruits of V. rotundifolia and its estrogen-like biological activity in human breast adenocarcinoma cells.Essential oil was extracted by SFE (supercritical fluid extraction) and its chemical composition was analysed by gas chromatography-mass spectrometry (GC-MS). The estrogenic activity of essential oil was evaluated in the MCF-7 cells by proliferation assay.The chemical composition of 40 compounds was determined and this represented 84.1 per cent of the total oil which was characterized by a high content of unsaturated (54.84% of total oil). The oil contained mainly linoleic (47.46%), (5.18%), untriacontane (2.28%), bicetyl (2.61%) and stearic (2.2%). The essential oil strongly stimulated the proliferation of MCF-7 cells, and this effect was significantly inhibited by the specific estrogen receptor antagonist ICI 182,780.Our results showed estrogen-like biological activity of the essential oil of the fruits from V. rotundifolia, and in future has a potential medical application.

Keyword: SCFA

Performance of an AnMBR pilot plant treating high-strength lipid wastewater: biological and filtration processes.

The performance of an anaerobic membrane bioreactor (AnMBR) treating wastewater with high levels of oil and grease content from a snacks factory is studied and its effectiveness is demonstrated. The relation between the reversible and the irreversible fouling rate and the fouling propensity of the matter were evaluated under a subcritical flux of 7.9 and 8.3 L/m(2) h. Low Oil and Grease (O&G) concentrations of 500 mg/L produced an irreversible fouling rate of only 0.09 mbar/d, while the fouling rate was between 0.96 and 3.95 mbar/d for an average O&G concentration of 6 g/L. In spite of the significant increase in filtration resistance from 0.31 to 6.08 × 10(12) m(-1) after 40 days of continuous operation, the critical flux level hardly decreased from 11.1 to 9.7 L/(m(2) h). With regard to the biological process, after a start-up period with an organic loading rate (OLR) of below 2 kg COD/(m(3) d), the system was able to treat wastewater between 4.6 and 36 g O&G/L and the system remained stable for OLR at around 17 kg COD/(m(3) d) for 2.8 d, without inhibitory signals. Acclimated sludge quickly reached maximum methane production and digested substrate with high oil and grease content, observing an increase in the first days and constant levels of propionic while were in the medium.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Flavor characteristic analysis of soymilk prepared by different soybean cultivars and establishment of evaluation method of soybean cultivars suitable for soymilk processing.

Flavor is an essential quality characteristics of soymilk, which contains compounds derived from via enzymatic and thermal reactions. In this study, 67 kinds of soybean cultivars were selected, and correlation analysis was conducted between physicochemical indexes of these soybean cultivars and flavor characteristic indexes of soymilk. With clustering analysis, all the soybean cultivars could be classified into three classes, and according to the results of principal component analysis for each class of soymilk flavor characteristics, the soymilk of second class had relatively heavier beany and non-beany flavor, and the third class had weaker flavor. For soybean cultivars of which the soymilk characteristics were unknown, two discriminant functions could be used to predict flavor characteristics if the physicochemical indexes were known. Therefore, screening of soybean cultivars suitable for soymilk processing can be targeted for the flavor favored by consumers and an evaluation method established.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Microelement contents and compositions of Rhus coriaria L. and Pistacia terebinthus L. fruits spread commonly in the south eastern Anatolia region of Turkey.

Sumac (Rhus coriaria L.) and terebinth (Pistacia terebinthus L.) are two important spice plants of south eastern Anatolia, Turkey. Nutrients, physical properties including moisture, ash, dry matter, protein, oil and essential oil content, along with Al, Ca, Cd, Cr, Cu, Fe, K, Mg, Na, Ni, Pb, Zn and characteristics of fruit sizes and compositions of both plants were determined from ripe fruits. The free content was determined in the fruit oil, and the main of sumac and terebinth were identified by gas chromatography/mass spectrometry. They included oleic, linoleic, and stearic in a ratio of 37.7% and 34.8%, 27.4% and 17.3%, 21.1% and 21.7% and, 4.7% and 2.5%, respectively. The fruits of both plants were rich in oil, and minerals, suggesting that they are valuable for using in food. The data may also be useful for the evaluation of nutritional information.

Keyword: SCFA

Model studies on release from different semisolid fat blends correlated with changes in sensory perception.

The effect of dispersed aqueous droplets in water-in-oil (W/O)-emulsion semisolid fats on aroma release and sensory perception was investigated on margarine models where model aroma substances were added. Aroma release from W/O-emulsion fat blends and bulk fat blends with added monoglycerides combining different of various short-chain free , methylketones, esters, and lactones were measured using headspace solid phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS), and their perception profiles were evaluated by sensory analysis. The presence of aqueous phase in a fat blend significantly reduced the headspace concentrations of butanoic and hexanoic , and also decreased the perceived intensity of total aroma and cheesy aroma. The aroma release of methylketones, esters, and lactones from the W/O-emulsion fat blends increased with increasing carbon chain length of the molecules. The intensity of aroma perception in a W/O-emulsion fat blend depended on the melting point of the (oleic, , stearic, and behenic) of the monoglyceride used as an emulsifier. Thus, aroma release from a W/O-emulsion semisolid fat blend was influenced by interactions between aroma volatiles and the dispersed aqueous droplets and by their viscoelastic properties.

Keyword: SCFA

Constituents of essential oil from the dried fruits and stems of Akebia quinata (THUNB.) DECNE.

The compositions of the essential oil from AKEBIAE FRUCTUS and AKEBIAE CAULIS, the dried fruits and stems of Akebia quinata (THUNB.) DECNE. (Lardizabalaceae), have been investigated by GC and GC/MS. As a result, the fruits oil was revealed the presence of 86 components, representing 98.4% of the total oil. The major compounds of the fruits oil were limonene, eugenol, octanal and p-cymene. The monoterpenoids and saturated short-chain aldehyde (C6 approximately C10) were main fractions of the oil. Ninety components accounting for 90.5% of constituents of stems oil were identified, and the main compounds of the oil were hexanoic , , (2E, 4E)-decadienal and hexanol. The oil had high content of saturated (C6 approximately C16), and unsaturated short-chain aldehyde (C6 approximately C10).

Keyword: SCFA

Substitution reactions of thorium(IV) acetate to synthesize nano-sized carboxylate complexes.

Some mixed-ligand thorium(IV) complexes with the general formula [Th(OOCCH(3))(4-n)L(n)] (L=anions of myristic, or stearic and n=1-4) have been synthesized by the stepwise substitution of acetate ions of thorium(IV) acetate with straight chain carboxylic in toluene under reflux. The complexes were characterized by elemental analyses, spectral (electronic, infrared, NMR and powder XRD) studies, electrical conductance and magnetic susceptibility measurements. Doubly and triply bridged coordination modes of the ligands were established by their infrared spectra and nano-size of the complexes by powder XRD. Room temperature magnetic susceptibility measurements revealed diamagnetic nature of the complexes. Electronic absorption spectra of the complexes showed pi-->pi*, n-->pi* and charge transfer transitions. Molar conductance values indicated the complex to be non-electrolytes. These are a new type of mixed-ligand thorium(IV) complexes for which a nano-sized, oxygen bridged polymeric structure has been established on the basis of physico-chemical studies.Copyright 2009 Elsevier B.V. All rights reserved.

Keyword: SCFA

Role of adiponectin in the protective action of dietary saturated fat against alcoholic liver in mice.

The protective effect of dietary saturated against the development of alcoholic liver disease has long been known, but the underlying mechanism is not completely understood. We examined the involvement of the adipocyte hormone adiponectin. Circulating adiponectin levels were significantly elevated by chronic ethanol administration to mice consuming a diet high in saturated fat. The increase in circulating adiponectin was associated with the activation a set of hepatic signaling pathways mediated through AMP-activated protein kinase, PPAR-alpha, and PPAR-gamma coactivator alpha, which in turn led to markedly increased rates of oxidation, prevention of hepatic steatosis, and alleviation of liver enzyme changes. Furthermore, treatment of rat 3T3-L1 adipocytes with saturated ( or stearic ) in the presence of ethanol increased secretion of adiponectin and enhanced activity of a mouse adiponectin promoter. In conclusion, the protective action of saturated fat against the development of alcoholic liver in mice is partially mediated through induction of adiponectin. The present findings suggest a novel paradigm for dietary in the pathogenesis of alcoholic liver disease and provide a promising therapeutic strategy-nutritional modulation of adiponectin-in treating human alcoholic liver disease.

Keyword: SCFA

Characterization of waxes used in pictorial artworks according to their relative amount of and hydrocarbons by gas chromatography.

A study attempted to characterize natural waxes used in pictorial works of art was carried out by means of gas chromatography. The analytical treatment requires prior hydrolysis of the waxes to release the (FA) (myristic (myr), (pal), oleic (ole), stearic (ste), araquidic (ara), behenic (beh), lignoceric (lig), cerotic (cer)) from the main esters of the waxes. The formation of derivatives of the was carried out by derivatization with ethyl choroformate (ECF). This derivatization reagent was chosen due to the speed, safety and quantitativity of the reaction. The analyzed hydrocarbons were n-eicosane, n-heneicosane, n-docosane, n-tricosane, n-tetracosane, n-pentacosane, n-hexacosane, n-heptacosane, n-octacosane, n-nonacosane, n-tricontane n-hentriacontane, n-dotriacontane, n-tritriacontane, n-tetratriacontane, n-pentatriacontane, main constituents of the waxes. No derivatization is needed to analyze the hydrocarbons. Ethyl ester derivatives and hydrocarbons are adequately separated by gas chromatography, identified by flame ionization detection and confirmed by mass spectrometry. To characterize natural waxes, peak area ratios of each with respect to the and peak area ratios of each hydrocarbons with respect to n-heptacosane were calculated. The proposed method provides a good characterization of different waxes most frequently used in artworks, such as beeswax, carnauba wax and ceresin, and has been successfully applied to real samples. This is the first report on the application of ECF to the analysis of in wax.

Keyword: SCFA

Properties of Oils From Plantain Pseudostem Biotransformed Using Crude Local Enzyme Sources: A Comparison of Poultry Feed Oil.

Plantain Pseudostem (PPS) wastes were biotransformed by applying Simultaneous Saccharification and Fermentation (SSF) using excised snail digestive juice and yeast slurry, and their oil properties compared with oils from commercially sold poultry feeds (PF). Patents suggesting the inclusion of certain additives (US20090226558A1), spices (US5741508A), cysteamine (US4711897A), and dextrin (US6326051B1) in animal diets are regarded as expensive, thus, requiring cheaper and readily available sources of growth.The analysis of their free radical scavenging potentials was carried by spectrophotometry, while , essentials oils, and phytosterols were determined by chromatography.After biotransformation, the melting point, specific gravity, , and peroxide values of the oils from SSF-PPS were significantly lower than those of PF, and showed elevations of C6:0- C18:1(trans-9) , , stearic, γ-Linolenic, α-linolenic, behenic, and lignoceric . Camphene, β-phelandrene, eugenol, β-elemene, bicyclogermacrene, guaiol, tetradecanoic , and hexadecanoic levels decreased when PPS was biotransformed. Lactic (1575.75 mg/100g), acetic (1234.26 mg/100g), propionic (845.74 mg/100g), and n-butyric (68.56 mg/100g) were the predominant (VFAs) in the SSF-PPS oils, which were higher than those found in the PF oil while PF oil contained higher campesterol, Stigmasterol, and 5-avesmasterol. The 2,2- azino-bis(3-ethylbenzthiazoline-6-sulphonic ) (ABTS), 1,1-diphenyl-2- picrylhydrazyl (DPPH), superoxide, and nitric oxide radical scavenging potentials of PPS oil at high concentrations after biotransformation were equivalent to the standards and the PF oil.This study has shown that biotransformation involving snail digestive juice and yeasts extensively improves the oil qualities of agricultural residues sufficient enough for poultry nutrition.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: SCFA

Eicosapentaenoic Potentiates Brown Thermogenesis through FFAR4-dependent Up-regulation of miR-30b and miR-378.

Emerging evidence suggests that n-3 polyunsaturated (PUFA) promote brown adipose tissue thermogenesis. However, the underlying mechanisms remain elusive. Here, we hypothesize that n-3 PUFA promotes brown adipogenesis by modulating miRNAs. To test this hypothesis, murine brown preadipocytes were induced to differentiate the of , oleate, or eicosapentaenoic (EPA). The increases of brown-specific signature genes and oxygen consumption rate by EPA were concurrent with up-regulation of miR-30b and 378 but not by oleate or . Next, we hypothesize that free receptor 4 (Ffar4), a functional receptor for n-3 PUFA, modulates miR-30b and 378. Treatment of Ffar4 agonist (GW9508) recapitulated the thermogenic activation of EPA by increasing oxygen consumption rate, brown-specific marker genes, and miR-30b and 378, which were abrogated in Ffar4-silenced cells. Intriguingly, addition of the miR-30b mimic was unable to restore EPA-induced Ucp1 expression in Ffar4-depleted cells, implicating that Ffar4 signaling activity is required for up-regulating the brown adipogenic program. Moreover, blockage of miR-30b or 378 by locked nucleic inhibitors significantly attenuated Ffar4 as well as brown-specific signature gene expression, suggesting the signaling interplay between Ffar4 and miR-30b/378. The association between miR-30b/378 and brown thermogenesis was also confirmed in fish oil-fed C57/BL6 mice. Interestingly, the Ffar4 agonism-mediated signaling axis of Ffar4-miR-30b/378-Ucp1 was linked with an elevation of cAMP in brown adipocytes, similar to cold-exposed or fish oil-fed brown fat. Taken together, our work identifies a novel function of Ffar4 in modulating brown adipogenesis partly through a mechanism involving cAMP activation and up-regulation of miR-30b and miR-378.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: SCFA

[Headspace solid-phase microextraction-gas chromatography-mass spectrometry for analysis of components from pollen pini].

Headspace solid-phase microextraction HS-SPME) technique was employed to extract the compounds from Pollen Pini. The components were isolated and identified successfully by gas chromatography-mass spectrometry (GC-MS). The volatiles were collected using several SPME parameters (including extraction time and temperature). The best response was obtained using a PA fiber by when the extraction temperature was 70 degrees C, extraction time was 30 minutes and desorption time was 10 time. Analysis was performed by GC-MS. Seventy components were identified by HS-SPME. The main components were compounds. The results showed that the HS-SPME technique could be used to extract the components from HS-SPME-GC-MS.

Keyword: SCFA

Modification and translocation of Rac/Rop guanosine 5\'-triphosphate-binding proteins of Scoparia dulcis in response to stimulation with methyl jasmonate.

Translocation of two Rac/Rop guanosine 5\'-triphosphate-binding proteins from Scoparia dulcis, Sdrac-1 and Sdrac-2, was examined employing transformed belladonna which overproduces these proteins as glutathione-S-transferase-tagged forms. The transferase activities of the fused proteins in microsomal fraction of belladonna markedly increased by the incubation with methyl jasmonate either in Sdrac-1 or Sdrac-2 transformant, while low and constant activities were observed in the untreated control. Recombinant Sdrac-2 protein was found to bind to prenyl chain in the presence of cell extracts prepared from methyl jasmonate-treated S. dulcis, however, Sdrac-1 was palmitoylated by the addition of the cell extracts. These results suggest that both Sdrac-1 and Sdrac-2 translocate to plant membranes by the stimulation with methyl jasmonate, however, targeting of these proteins is triggered by the independent modification mechanisms, palmitoylation for Sdrac-1 and prenylation for Sdrac-2.

Keyword: SCFA

An inhibitor of phospholipase A2 group IIA modulates adipocyte signaling and protects against diet-induced metabolic syndrome in rats.

Obesity, type 2 diabetes, and cardiovascular disease correlate with infiltration to adipose tissue of different immune cells, with uncertain influences on metabolism. Rats were fed a diet high in carbohydrates and saturated fats to develop diet-induced obesity over 16 weeks. This nutritional overload caused overexpression and secretion of phospholipase A(2) group IIA (pla2g2a) from immune cells in adipose tissue rather than adipocytes, whereas expression of adipose-specific phospholipase A(2) (pla2g16) was unchanged. These immune cells produce prostaglandin E(2) (PGE(2)), which influences adipocyte signaling. We found that a selective inhibitor of human pla2g2a (5-(4-benzyloxyphenyl)-(4S)-(phenyl-heptanoylamino)-pentanoic [KH064]) attenuated secretion of PGE(2) from human immune cells stimulated with the , , or with lipopolysaccharide. Oral administration of KH064 (5 mg/kg/day) to rats fed the high-carbohydrate, high-fat diet prevented the overexpression of pla2g2a and the increased macrophage infiltration and elevated PGE(2) concentrations in adipose tissue. The treatment also attenuated visceral adiposity and reversed most characteristics of metabolic syndrome, producing marked improvements in insulin sensitivity, glucose intolerance, and cardiovascular abnormalities. We suggest that pla2g2a may have a causal relationship with chronic adiposity and metabolic syndrome and that its inhibition in vivo may be a valuable new approach to treat obesity, type 2 diabetes, and metabolic dysfunction in humans.

Keyword: SCFA

Time course effects of fermentation on and compound profiles of Cheonggukjang using new soybean cultivars.

In this study, we investigated the effects of the potential probiotic Bacillus subtilis CSY191 on the profiles of Cheonggukjang, a fermented soybean paste, prepared using new Korean brown soybean cultivars, protein-rich cultivar (Saedanbaek), and oil-rich cultivar (Neulchan). Twelve were identified in the sample set-myristic, , palmitoleic, stearic, oleic, vaccenic, linoleic, α-linolenic, arachidic, gondoic, behenic, and lignoceric -yet, no specific changes driven by fermentation were noted in the profiles. To further explore the effects of fermentation of B. subtilis CSY191, complete profiles of volatiles were monitored. In total, 121, 136, and 127 compounds were detected in the Saedanbaek, Daewon (control cultivar), and Neulchan samples, respectively. Interestingly, the content of pyrazines-compounds responsible for pungent and unpleasant Cheonggukjang flavors-was significantly higher in Neulchan compared to that in Saedanbaek. Although the fermentation period was not a strong factor affecting the observed changes in profiles, we noted that profiles of volatiles in Cheonggukjang changed significantly over time, and different cultivars represented specific profiles. Thus, further sensory evaluation might be needed to determine if such differences influence consumers\' preferences. Furthermore, additional studies to elucidate the associations between B. subtilis CSY191 fermentation and other nutritional components (e.g., amino ) and their health-promoting potential are warranted.Copyright © 2016. Published by Elsevier B.V.

Keyword: SCFA

The acetate recovery factor to correct tracer-derived dietary fat oxidation in humans.

When using (13)C tracer to measure plasma fat oxidation, an acetate recovery factor should be determined in every subject to correct for label sequestration. Less is known regarding the acetate recovery factor for dietary oxidation. We compiled data from six studies to investigate the determinants of the dietary acetate recovery factor (dARF) at rest and after physical activity interventions and compared the effects of different methods of dARF calculation on both the fat oxidation and its variability. In healthy lean subjects, dARF was 50.6 +/- 5.4% dose (n = 56) with an interindividual coefficient of variation of 10.6% at rest and 9.2% after physical activity modifications. The physical activity interventions did not impact dARF, and the intraindividual coefficient of variation was 4.6%. No major anthropological or physiological determinants were detected except for resting metabolic rate, which explains 7.4% of the dARF variability. Applying an individual or an average group dARF did not affect the mean and the variability of the derived dietary lipid oxidation at rest or after physical activity interventions. Using a mean dARF for a group leads to over- or underestimation of fat oxidation of less than 10% in individual subjects. Moreover, the use of a group or individual correction did not affect the significant relationship found between fasting respiratory exchange ratio and dietary fat oxidation. These data indicate that an average dARF can be applied for longitudinal and cross-sectional studies investigating dietary lipid metabolism.

Keyword: SCFA

Metabolic signatures and risk of type 2 diabetes in a Chinese population: an untargeted metabolomics study using both LC-MS and GC-MS.

Metabolomics has provided new insight into diabetes risk assessment. In this study we characterised the human serum metabolic profiles of participants in the Singapore Chinese Health Study cohort to identify metabolic signatures associated with an increased risk of type 2 diabetes.In this nested case-control study, baseline serum metabolite profiles were measured using LC-MS and GC-MS during a 6-year follow-up of 197 individuals with type 2 diabetes but without a history of cardiovascular disease or cancer before diabetes diagnosis, and 197 healthy controls matched by age, sex and date of blood collection.A total of 51 differential metabolites were identified between cases and controls. Of these, 35 were significantly associated with diabetes risk in the multivariate analysis after false discovery rate adjustment, such as increased branched-chain amino (leucine, isoleucine and valine), non-esterified (, stearic , oleic and linoleic ) and lysophosphatidylinositol (LPI) species (16:1, 18:1, 18:2, 20:3, 20:4 and 22:6). A combination of six metabolites including proline, glycerol, aminomalonic , LPI (16:1), 3-carboxy-4-methyl-5-propyl-2-furanpropionic and urea showed the potential to predict type 2 diabetes in at-risk individuals with high baseline HbA1c levels (≥6.5% [47.5\xa0mmol/mol]) with an AUC of 0.935. Combined lysophosphatidylglycerol (LPG) (12:0) and LPI (16:1) also showed the potential to predict type 2 diabetes in individuals with normal baseline HbA1c levels (<6.5% [47.5\xa0mmol/mol]; AUC\u2009=\u20090.781).Our findings show that branched-chain amino and NEFA are potent predictors of diabetes development in Chinese adults. Our results also indicate the potential of lysophospholipids for predicting diabetes.

Keyword: SCFA

Recycling of food waste for fuel precursors using an integrated bio-refinery approach.

The main aim was to integrate FW-recycling with cultivation of Rhodotorula glutinis and anaerobic digestion (AD) for bio-energy and -fuel recovery. Mixed FW was mechanically macerated (Pcon) and hydrolysed (at 250gL water) via chemical (Ch), thermal (Th) and TCh (combined Ch and Th) treatments. Cleared hydrolysates from individual pre-treatment processes were used as culture medium for cultivation of R. glutinis, while the residual solids (RS) were subjected to AD. Pcon cultivation yielded maximal R. glutinis dry weight biomass (5.18gL) and total fatty contents (1.03gg DW). Dominant fatty methyl esters (FAME) were - (C-26%); Stearic - (C-17%) and Oleic acids (C-38%), ideal for bio-diesel production. Highest methane yields (actual ∼0.139mkg volatile solids) were measured from AD of Th-derived RS. Thus integrated FW recycling approaches will be more feasible for generating energy and economic incentives.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Physicochemical and volatiles characterization of trans-free solid fats produced by lipase-catalyzed interesterification.

Trans-free solid fats were synthesized from fully hydrogenated soybean oil (FHSBO), olive oil (OO), and palm stearin (PS) at different substrate weight ratios (10:20:70, 10:40:50 and 10:50:40) via lipase-catalyzed interesterification. The interesterified products contained mostly TAG (98.8% to 99.0%), and small amounts of MAG and DAG as by-products. The major were oleic , , and stearic in the interesterified products, and the melting points ranged from 39 to 45 degrees C. The amount of alpha-tocopherol was reduced by 75% to 92%. analysis by solid-phase microextraction indicated that OO and PS had distinct profiles, in which 18 volatiles were retained in interesterified products. Furthermore, some volatiles disappeared or formed during processing. Electronic nose showed that the odors of substrates (OO and PS) were different from each other, and the odors of interesterified products were distinguishable from that of OO or PS. Among the interesterified products, the odor of blend FHSBO:OO:PS of 10:40:50 or 10:50:40 was different from that of blend FHSBO:OO:PS (10:20:70). However, no odor difference was observed between products blend FHSBO:OO:PS 10:40:50 and 10:50:40.

Keyword: SCFA

Short- and medium-chain exhibit antimicrobial activity for oral microorganisms.

This study assessed the antibacterial activity of short-, medium-, and long-chain against various oral microorganisms.The short-chain [formic (C1), acetic (C2), propionic (C3), butyric (C4), isobutyric (C4), isovaleric (C5), hexanoic (C6)], medium-chain [octanoic (C8), capric (C10), lauric (12)], and long-chain [myristic (C14), (C16)], were investigated for antimicrobial activity against Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguis, Candida albicans, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis.The data demonstrated that the exhibited patterns of inhibition against oral bacteria with some specificity that appeared related more to the bacterial species that the general structural characteristics of the microorganism. As a group the were much less effective against C. albicans than the oral bacteria, with effectiveness limited to hexanoic, octanoic, and lauric . Formic , capric, and lauric were broadly inhibitory for the bacteria. Interestingly, that are produced at metabolic end-products by a number of these bacteria, were specifically inactive against the producing species, whilst substantially inhibiting the growth of other oral microorganisms.The results indicate that the antimicrobial activity of short-chain (SCFAs), medium-chain (MCFAs), long-chain (LCFAs) could influence the microbial ecology in the oral cavity via at least 2 potential pathways. First, the agents delivered exogenously as therapeutic adjuncts could be packaged to enhance a microbial-regulatory environment in the subgingival sulcus. Second, it would be the intrinsic nature of these inhibitors in contributing to the characteristics of the microbial biofilms, their evolution, and emergence of species within the biofilms. Further studies on these functions are required to better understand the nature of these potential microbial interactions in the biofilms.Copyright © 2011 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Effect of co-culture with Tetragenococcus halophilus on the physiological characterization and transcription profiling of Zygosaccharomyces rouxii.

Zygosaccharomyces rouxii and Tetragenococcus halophilus are widely existed and play vital roles during the manufacture of fermented foods such as soy sauce. The aim of this study was to elucidate the effect of T. halophilus CGMCC 3792 on the physiological characterizations and transcription profiling of Z. rouxii CGMCC 3791. Salt tolerance analysis revealed that co-culture with T. halophilus enhanced the salt tolerance of Z. rouxii during salt stress. Analysis of the compounds revealed that co-culture reduced the level of 1-butanol, improved the level of octanoic which all were produced by T. halophilus and reduced the level of phenylethyl alcohol produced by Z. rouxii. The presence of Z. rouxii decreased the contents of 3,4-dimethylbenzaldehyde and acetic produced by T. halophilus. In addition, co-culture improved the content of benzyl alcohol significantly. Analysis of membrane showed that co-culture improved the content of (C16:0) and stearic (C18:0) in cells of Z. rouxii, and reduced the contents of myristic (C14:0), palmitoleic (C16:1) and oleic (C18:1). In order to further explore the interactions between the two strains, RNA-seq technology was used to investigate the effect of co-culture with T. halophilus on the transcription profiling of Z. rouxii. By comparing cells incubated in co-culture group with cells incubated in single-culture group, a total of 967 genes were considered as differentially expressed genes (DEGs). Among the DEGs, 72 genes were up-regulated, while 895 genes were down-regulated. These DEGs took party in various activities in cells of Z. rouxii, and the result showed co-culture with T. halophilus had a positive effect on proteolysis, the attachment of a cell to another cell, extracellular protein accumulation, energy metabolism, and a negative effect on oxidative phosphorylation, small molecular substances metabolism, DNA replication and repair, and transcription in cells of Z. rouxii. Results presented in this study may contribute to further understand the interactions between Zygosaccharomyces rouxii and Tetragenococcus halophilus.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Influence of feeding whole sunflower seed and extruded linseed on production of dairy cows, rumen and plasma constituents, and composition of milk.

Holstein cows were fed total mixed rations (TMR) supplemented with protected palm fat (PPF), whole sunflower seed (WSS) or extruded linseed (ELS) for 100 days. Percentage of dietary crude fat was 5.3, 5.1 and 5.1, respectively. Diet had no (p > 0.05) effect on feed intake, milk yield or milk protein content. Percentage of milk fat and yield of fat--corrected milk were significantly increased when diets were supplemented with WSS and ELS. Feeding PPF resulted in the lowest (p < 0.05) ruminal concentration of . No significant dietary effect on plasma characteristics was observed. Concentration of polyunsaturated (PUFA) was higher (p < 0.05), and PUFA n-6/n-3 ratio lower (p < 0.05), in the milk fat from cows fed ELS compared to WSS. Supplementation of TMR with oilseeds compared to PPF increased the content of CLA in milk fat (p < 0.005) and decreased its atherogenicity, primarily due to a significant reduction of concentration. Both oilseeds significantly improved the spreadability index of manufactured butter. ELS, but not WSS, increased the susceptibility of milk fat to oxidation (p < 0.05). It can be concluded that feeding of oilseeds to dairy cows improved nutritional quality of milk fat, with supplementation with ELS producing an even more desirable milk profile than WSS supplementation.

Keyword: SCFA

Obesity changes the human gut mycobiome.

The human intestine is home to a diverse range of bacterial and fungal species, forming an ecological community that contributes to normal physiology and disease susceptibility. Here, the fungal (mycobiome) in obese and non-obese subjects was characterized using Internal Transcribed Spacer (ITS)-based sequencing. The results demonstrate that obese patients could be discriminated by their specific fungal composition, which also distinguished metabolically "healthy" from "unhealthy" obesity. Clusters according to genus abundance co-segregated with body fatness, fasting triglycerides and HDL-cholesterol. A preliminary link to metabolites such as hexadecanedioic , caproic and N-acetyl-L-glutamic was also found. Mucor racemosus and M. fuscus were the species more represented in non-obese subjects compared to obese counterparts. Interestingly, the decreased relative abundance of the Mucor genus in obese subjects was reversible upon weight loss. Collectively, these findings suggest that manipulation of gut mycobiome communities might be a novel target in the treatment of obesity.

Keyword: SCFA

Effect of conjugated linoleic and acetate on milk fat synthesis and adipose lipogenesis in lactating dairy cows.

During biohydrogenation-induced milk fat depression (MFD), nutrients are spared from milk fat synthesis and are available for other metabolic uses. Acetate is the major carbon source spared and it may increase lipid synthesis in adipose tissue during MFD. The objective of this study was to compare the effect of trans-10,cis-12 conjugated linoleic (CLA) and the amount of acetate spared during CLA-induced MFD on adipose tissue lipogenesis. Nine multiparous, lactating, ruminally cannulated Holstein cows (244 ± 107 d in milk; 25 ± 8.4 kg of milk/d; mean ± standard deviation) were randomly assigned to treatments in a 3 × 3 Latin square design. Experimental periods were 4 d followed by a 10-d washout. Treatments were control (CON), ruminal infusion of acetate (AC; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), or abomasal infusion of CLA (10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA). Dry matter intake, milk yield, and milk protein yield and percentage were not affected by treatments. Compared with CON, milk fat yield decreased 23% and fat percent decreased 28% in CLA, and milk fat yield increased 20% in AC. Concentration and yield of milk de novo synthesized (C16) was increased by CLA, compared with CON. Yield of de novo synthesized and was increased by AC, compared with CON. Lipogenesis capacity of adipose tissue explants was decreased 72% by CLA, but was not affected by AC. Acetate oxidation by adipose explants was not affected by treatments. Treatments had no effect on expression of key lipogenic factors, lipogenic enzymes, and leptin; however, expression of binding protein 4 was reduced in CLA compared with CON. Additionally, hormone-sensitive lipase and perilipin 1 were decreased by CLA and acetate. Plasma glucose and glucagon concentrations were not affected by treatments; however, CLA increased nonesterified 17.7%, β-hydroxybutyrate 16.1%, and insulin 27.8% compared with CON, and AC increased plasma β-hydroxybutyrate 18%. In conclusion, during CLA-induced MFD in low-producing cow adipose tissue was sensitive to the anti-lipogenic effects of CLA, while spared acetate did not stimulate adipose lipogenesis. However, acetate may play an important role in stimulating lipogenesis and improving energy status in the mammary gland under normal conditions.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Co-overexpression of native phospholipid-biosynthetic genes plsX and plsC enhances lipid production in Synechocystis sp. PCC 6803.

The overexpression of native plsX and plsC genes involving in /phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and oxygen evolution rate of all overexpressing (OX) strains were normally similar as wild type. For compositions, saturated , in particular (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated were observed, specifically linoleic (18:2) and alpha-linolenic (18:3). The plsC/plsX-overexpressing (OX\u2009+\u2009XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX\u2009+\u2009XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.

Keyword: SCFA

Ailanthus altissima (Miller) Swingle seed oil: chromatographic characterization by GC-FID and HS-SPME-GC-MS, physicochemical parameters, and pharmacological bioactivities.

This study aimed to identify the physicochemical and the chemical properties of Ailanthus altissima (Miller) Swingle seed oil and to evaluate its in vitro antioxidant and antibacterial activities and in vivo analgesic and anti-inflammatory activities. The \' composition was determined using GC-FID. The oil was screened for antioxidant activity by DPPH test. The analgesic and anti-inflammatory activities were determined using the acetic writhing test in mice and the carrageenan-induced paw edema assay in rats, respectively. compounds were characterized by HS-SPME-GC-MS. A. altissima produces seeds which yielded 17.32% of oil. The seed oil was characterized by a saponification number of 192.6\xa0mg KOH∙g of oil, a peroxide value of 11.4\xa0meq O∙kg of oil, a K of 4.04, a K of 1.24, and a phosphorus content of 126.2\xa0ppm. The main identified were (3.06%), stearic (1.56%), oleic (38.35%), and linoleic ones (55.76%). The main aroma compounds sampled in the headspace were carbonyl derivatives. The oil presents an important antioxidant activity (IC\u2009=\u200924.57\xa0μg/mL) and a modest antimicrobial activity. The seed oil at 1\xa0g/kg showed high analgesic (91.31%) and anti-inflammatory effects (85.17%). The presence of high levels of unsaturated and the noteworthy antioxidant capacity of the seed oil can hypothesize its use as an analgesic and anti-inflammatory agent.

Keyword: SCFA

Physico-chemical characteristics of papaya (Carica papaya L.) seed oil of the Hong Kong/Sekaki variety.

A study was carried out to determine the physicochemical characteristics of the oil derived from papaya seeds of the Hong Kong/Sekaki variety. Proximate analysis showed that seeds of the Hong Kong/Sekaki variety contained considerable amount of oil (27.0%). The iodine value, saponification value, unsaponifiable matter and free contents of freshly extracted papaya seed oil were 76.9 g I2/100g oil, 193.5 mg KOH/g oil, 1.52% and 0.91%, respectively. The oil had a Lovibond color index of 15.2Y + 5.2B. Papaya seed oil contained ten detectable , of which 78.33% were unsaturated. Oleic (73.5%) was the dominant followed by (15.8%). Based on the high performance liquid chromatography (HPLC) analysis, seven species of triacylglycerols (TAGs) were detected. The predominant TAGs of papaya seed oil were OOO (40.4%), POO (29.1%) and SOO (9.9%) where O, P, and S denote oleic, and stearic , respectively. Thermal analysis by differential scanning calorimetry (DSC) showed that papaya seed oil had its major melting and crystallization transitions at 12.4°C and -48.2°C, respectively. Analysis of the sample by Z-nose (electronic nose) instrument showed that the sample had a high level of compounds.

Keyword: SCFA

The influence of decreased hydraulic retention time on the performance and stability of co-digestion of sewage sludge with grease trap sludge and organic fraction of municipal waste.

The effect of hydraulic retention time ranging from 12 to 20\xa0d on process performance and stability was investigated in two anaerobic completely stirred tank reactors with a working liquid volume equal to 6 litres. The reactors were fed with mixtures containing (on solids basis): 40% of sewage sludge, 30% of organic fraction of municipal waste and 30% of grease trap sludge. The change of hydraulic retention time did not significantly affect process stability. However, methane yields as well as solids removal decreased from 0.54 to 0.47\xa0l per kg of added solids and 65% to 60% respectively, with the decrease of hydraulic retention time. Despite the fact that the best process performance was achieved for hydraulic retention time of 20 days, the obtained results showed that it is also possible to carry out the co-digestion process at shorter hydraulic retention times with good results. Furthermore, gas production rate as well as biogas production at the shortest hydraulic retention time were approximately 46% higher in comparison to results obtained at the longest hydraulic retention time. In this context, the proposed solution seems to be an interesting option, because it provides an unique opportunity for wastewater treatment plants to improve their profitability by enhancing energy recovery from sludge as well as full utilisation of the existing infrastructure and hence creates a new potential place for alternative treatment of organic industrial waste such as: fat-rich materials or food waste. However, implementation of the solution at wastewater treatment plants is still a big challenge and needs studies including identification of optimal digesting conditions, information about substrate pumping, inhibition thresholds and processing properties. Additionally, due to the characteristics of both co-substrates their introduction to the full-scale digester should be carefully planned due to a potential risk of overloading of the digester. For this reason, a gradual increase of the share of these wastes in the co-digestion mixture is highly recommended, because it will allow for the acclimatization of bacteria as well as prevent overloading. The results of this study show the importance of gradual acclimatization of microorganisms to the changing environmental conditions. It was found that concentration of long chain in effluents increased with the reduction of hydraulic retention time, but this phenomenon did not significantly influence the performance and stability of the process probably due to changes hydraulic retention time being gradual. Although for a moderate negative correlation with solids removal was observed.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Pulverizing processes affect the chemical quality and thermal property of black, white, and green pepper ( L.).

In this study, the effects of different pulverizing methods on the chemical attributes and thermal properties of black, white and green pepper were evaluated. Cryogenic grinding minimally damaged the lipid, moisture, crude protein, starch, non- ether extract, piperine, essential oil and the typical pepper essential oil compounds of the spices. The pulverizing methods and storage significantly affected the compositions of the in the peppers, except for and lignoceric . The amino contents and the thermo-gravimetric analysis curve were hardly influenced by the grinding techniques. The use of cryogenic grinding to prepare pepper ensured the highest quality of pepper products. Regardless of grinding technique, the values of moisture, piperine, unsaturated , essential oil, monoterpenes, and the absolute concentrations of typical pepper essential oil constituents (except caryophyllene oxide) decreased, whereas the amino , lipid, protein, starch, and non- ether extract content as well as the thermal properties were insignificantly changed after storage at 4\xa0°C for 6\xa0months.

Keyword: SCFA

Identification and characterization of PPARα ligands in the hippocampus.

Peroxisome proliferator-activated receptor-α (PPARα) regulates hepatic fatty catabolism and mediates the metabolic response to starvation. Recently we found that PPARα is constitutively activated in nuclei of hippocampal neurons and controls plasticity via direct transcriptional activation of CREB. Here we report the discovery of three endogenous PPARα ligands-3-hydroxy-(2,2)-dimethyl butyrate, hexadecanamide, and 9-octadecenamide-in mouse brain hippocampus. Mass spectrometric detection of these compounds in mouse hippocampal nuclear extracts, in silico interaction studies, time-resolved FRET analyses, and thermal shift assay results clearly indicated that these three compounds served as ligands of PPARα. Site-directed mutagenesis studies further revealed that PPARα Y464 and Y314 are involved in binding these hippocampal ligands. Moreover, these ligands activated PPARα and upregulated the synaptic function of hippocampal neurons. These results highlight the discovery of hippocampal ligands of PPARα capable of modulating synaptic functions.

Keyword: SCFA

Synthesis of lipophilic 2-oxoamides based on gamma-aminobutyric and delta-aminovaleric analogues and their activity against phospholipase A2.

A variety of lipophilic 2-oxoamides based on gamma-aminobutyric and delta-aminovaleric analogues were synthesized. 2-oxoamides containing a tetrazole, a thioethyl or a thioacetyl group are weak inhibitors of GIVA cPLA(2), while derivatives containing a methyl tetrazole, a diethyl phosphonate or a thioethyl group are weak inhibitors of GV sPLA(2).Copyright (c) 2007 European Peptide Society and John Wiley & Sons, Ltd.

Keyword: SCFA

profile of the milk of cows reared in the mountain region of Poland.

An appropriate composition of milk (FA) improves the nutritional value of milk and milk products, and improves milk processing. Polish dairy farms in the mountainous region are rather small, animal nutrition there is based on locally produced forages and this, together with the transitional climate zone brings about seasonal changes in FA composition of milk. The aim of the study was to evaluate the composition of FA in bovine milk fat in relation to fat intake in forages and their FA profiles. The study involved 5 herds reared in low-input mountain farms located at an altitude of 670-780 m above sea level (Beskid Mountains). The cows were fed forages produced locally. FAs in forages and milk samples were subjected to gas chromatography. Highest fat intake observed in grazing season (4·2-4·7%) and high amounts of polyunsaturated FA in forages from that period (51·8-64·1 g/100 g FA) resulted in a markedly high content of valuable FAs: t-11 C18:1 (3·22 g/100 g FA), c-9, t-11 C18:2 (CLA; 1·20 g/100 g FA) in milk. Lower fat intake of forages containing high amount of SFA (32·42-38·83 g/100 g FA) in the indoor period resulted in changes in milk composition. The content of total short-chain saturated FA () was highest in winter and early spring samples (14·10 and 13·44 g/100 g FA, respectively), like the amounts of myristic C14:0 and C16:0 (11·80 and 37·92 g/100 g FA). Total odd- and branched-chain (OBCFA; 6·58 g/100 g FA) content was highest at the beginning of the grazing period. Fresh grass consumed by cows promoted the activity of Δ(9)-desaturase in mammary gland as evidenced by higher C14:1 : C14:0 (0·054) and C16:1 : C16:0 (0·026) ratios in grazing than in the indoor periods.

Keyword: SCFA

Automated production of [11C]acetate and [11C]palmitate using a modified GE Tracerlab FX(C-Pro).

As researchers explore new applications for positron emission tomography radiopharmaceuticals, the demand for effective and readily available radiopharmaceuticals continues to increase. The syntheses of two such radiopharmaceuticals, [(11)C]acetate and [(11)C]palmitate, can be automated on the GE Tracerlab FX(C-Pro) by utilizing Grignard reactions. Radiochemical purities of the [(11)C]acetate and the [(11)C]palmitate products were high (>98% and >99.9%, respectively) with average non-corrected yields of 18% (n = 3) and 10% (n = 5), respectively. These data comprise the validation trials for site qualification of clinical production of both radiopharmaceuticals.Copyright © 2011 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Regulation of peroxisome proliferator-activated receptor gamma on milk fat synthesis in dairy cow mammary epithelial cells.

Peroxisome proliferator-activated receptor gamma (PPARγ) participates in lipogenesis in rats, goats, and humans. However, the exact mechanism of PPARγ regulation on milk fat synthesis in dairy cow mammary epithelial cells (DCMECs) remains largely unexplored. The aim of this study was to investigate the role of PPARγ regarding milk fat synthesis in DCMECs and to ascertain whether milk fat precursor acetic and could interact with PPARγ signaling to regulate milk fat synthesis. For this study, we examined the effects of PPARγ overexpression and gene silencing on cell growth, triacylglycerol synthesis, and the messenger RNA (mRNA) and protein expression levels of genes involved in milk fat synthesis in DCMECs. In addition, we investigated the influences of acetic and on the mRNA and protein levels of milk lipogenic genes and triacylglycerol synthesis in DCMECs transfected with PPARγ small interfering RNA (siRNA) and PPARγ expression vector. The results showed that when PPARγ was silenced, cell viability, proliferation, and triacylglycerol secretion were obviously reduced. Gene silencing of PPARγ significantly downregulated the expression levels of milk fat synthesis-related genes in DCMECs. PPARγ overexpression improved cell viability, proliferation, and triacylglycerol secretion. The expression levels of milk lipogenic genes were significantly increased when PPARγ was overexpressed. Acetic and could markedly improve triacylglycerol synthesis and upregulate the expression levels of PPARγ and other lipogenic genes in DCMECs. These results suggest that PPARγ is a positive regulator of milk fat synthesis in DCMECs and that acetic and could partly regulate milk fat synthesis in DCMECs via PPARγ signaling.

Keyword: SCFA

Fast ultrasound-assisted extraction of polar (phenols) and nonpolar (lipids) fractions in Heterotheca inuloides Cass.

Heterotheca inuloides Cass., also known as "arnica", is used in traditional medicine in Mexico.Development of fast methods for the extraction of lipidic and phenolic fractions from arnica plants and their subsequent characterization.Ultrasound was applied to accelerate extraction of the target compounds from this plant and reduce the use of organic solvents as compared with conventional methods. Gas chromatography-ion trap mass spectrometry and liquid chromatography with diode-array detection were used for the characterization of the lipidic and phenolic fractions, respectively.Under optimal extraction conditions, 9 and 55\u2009min were necessary to complete extraction of the lipidic and phenolic fractions, respectively. The present at the highest concentrations in H. inuloides were eicosatetraenoic n3 (24.6\u2009μg/g), cis-9-hexadecenoic n7 (23.1\u2009μg/g), exacosanoic (22.7\u2009μg/g) and cis-9-octadecenoic (21.3\u2009μg/g), while the rest were in the range 7.6-1.3\u2009μg/g. The most concentrated phenols were guaiacol (41.5\u2009μg/g), catechin (38.7\u2009μg/g), ellagic (35.9\u2009μg/g), carbolic (24.2\u2009μg/g) and p-coumaric (19.5\u2009μg/g), while the rest were in the range 5.1-0.4\u2009μg/g.Ultrasound reduces the time necessary to complete the extraction 160 and 26 times, the extraction volume 2.5 and 4 times, and increases the extraction efficiency 5 and 3 times for lipidic and phenolic fractions, respectively, in comparison with conventional extraction methods. In addition, the characterization of the lipidic and phenolic fractions constitutes a first approach to the H. inuloides metabolome.Copyright © 2011 John Wiley & Sons, Ltd.

Keyword: SCFA

Impact of Microclimate on and Terpenes in "Kerman" and "Golden Hills" Pistachio (Pistacia vera) Kernels.

Pistachio is an economically important nut crop in California. Since temperature variations among geographical locations can influence biochemical processes during fruit development, it is of great relevance to understand the impact of growing area over the components that define the nutritional and sensory characteristics of pistachio nuts. Changes in moisture, fat content, composition and terpenes were studied during kernel development for "Kerman" and "Golden Hills" varieties in two different California Central Valley microclimates, Lost Hills and Parlier. Moisture content decreased from July to September for both cultivars at both locations. Kerman had a higher moisture content at both locations compared with Golden Hills. Harvest time affected fat content only for Kerman, where the values increased drastically from 21-July to 4-Aug, then remained constant. Golden Hills\' fat content remained constant during the period of the study. The main in pistachio oil is oleic (46% to 59%), followed by linoleic (26% to 36%) and (11% to 16%). C16:0, C16:1, C18:2, and C18:3 decreased with harvest time, while C18:1 increased. α-Pinene was the most concentrated among the cultivars and locations. It decreased with harvest time for both cultivars at both locations, ranging from 105 to 2464 mg/kg. At harvest, Golden Hills and Kerman at Parlier both had higher concentrations of α-pinene than the two cultivars at Lost Hills. Our results demonstrate that microclimate affects biosynthesis of and terpenes in pistachio kernels, the main compounds responsible for pistachio nutritional and sensory characteristics.© 2019 Institute of Food Technologists®.

Keyword: SCFA

Cypate and Cypate-Glucosamine as Near-Infrared Fluorescent Probes for In Vivo Tumor Imaging.

Near-infrared (NIR) imaging is a promising technique for use as a noninvasive and sensitive diagnostic tool. Although the NIR fluorescently labeled glucose analog glucosamine (cypate-glucosamine) has applications in preclinical imaging, the transport pathways and fate of this probe in tissues remain unaddressed. Here, we have synthesized and characterized cypate and cypate-glucosamine conjugate (cy-2-glu), and investigated the probable transport pathways of these probes in vitro and in vivo. We compared uptake of the probes in the presence and absence of excess d-glucose, "saturated cypate" and in two normal-cancer cell line pairs: lung cancer (A549)-normal (MRC9) and prostate cancer (DU145)-normal (BPH). Breast cancer (MDA-MB-231) and liver cancer (HepG2) cell lines were also examined. Results support use of the glucose transport pathway by cy-2-glu and transport pathway by cypate. Mass spectrometry data on the in vitro extracts revealed deamidation of cy-2-glu in prostate and liver cells, suggesting release of glucosamine. In vivo biodistribution studies in mice engrafted with breast tumors showed a distinct accumulation of cy-2-glu in liver and tumors, and to a lesser extent in kidneys and spleen. A negligible accumulation of cypate alone in tumors was observed. Analysis of urine extracts revealed renal excretion of the cy-2-glu probe in the form of free cypate, indicating deamidation of cy-2-glu in tissues. Thus, investigation of the metabolic pathways used by NIR probes such as cy-2-glu advances their use in the detection and monitoring of tumor progression in preclinical animal studies.Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: SCFA

Nutritional composition of frog (Rana esculanta) waste meal.

In the present study, the waste obtained from the frozen frog leg industry was used for the production of frog waste meal, and its proximate, amino , , mineral and vitamin compositions were evaluated to determine the nutritional quality. In addition, the total bacterial count, Salmonella, total basic nitrogen (TVB-N, mgN/100g) and thiobarbituric (TBA, mg malonaldehyde/kg) were also measured to determine the microbiological and chemical quality of frog waste meal (FWM). The crude protein, fat and ash content of FWM on a dry weight basis were 68.6%, 17.0% and 13.2%, respectively. The amino profiles were found to be fairly close to those of fish meal in terms of protein sources and rich in the glutamic , glycine, proline, arginine, and methionine. The proportions of composition in FWM were analysed and findings were 26.7% for total saturated (SFA), 42.5% for total monounsaturated (MUFA), 17.0% for total n - 6 and 3.3% for n - 3 ratio. The major SFA, MUFA and PUFA in FWM were (19.1%), oleic (26.0%) and linoleic (16.7%), respectively. FWM was found to be high in mineral content, especially Zn, K, Cu, Mn, and Mg and high level of some vitamins such as folic and thiamin. The total bacterial count was found to be 2.9x10(4) CFU/g, and Salmonella was not observed. TVB-N and TBA in FWM was determined to be 157.4+/-5.8 mg N/100g and 1.2+/-0.1 mg malonaldehyde/kg, respectively.

Keyword: SCFA

GPR40 activation leads to CREB and ERK phosphorylation in primary cultures of neurons from the mouse CNS and in human neuroblastoma cells.

GPR40, the free receptor 1, is expressed strongly in the primate pancreas and brain. While the role of pancreatic GPR40 in glucose homeostasis has been extensively studied, the absence of this G-protein-coupled receptor from the brain of rodents has hampered studies into its role in the central nervous system. However, we found intense GPR40 mRNA expression by in situ hybridization in mouse hippocampal and motor cortex neurons. Furthermore, in a neuroblastoma cell GPR40 was activated by docosahexaenoic and selective agonists, yet not by . Significantly, the activation of GPR40 provoked the phosphorylation of the cAMP response element-binding protein, CREB. The receptor was also functional in primary cultures of murine neurons, in which its activation by a selective agonist produced the phosphorylation of CREB and of extracellular signal-regulated kinases, ERK1/2. These results suggest that mice represent a suitable model for elucidating the role of GPR40 in brain function.© 2014 Wiley Periodicals, Inc.

Keyword: SCFA

Free and their esters modulate isothermal crystallization of anhydrous milk fat.

The effect of free with different chain lengths or unsaturation degree on anhydrous milk fat (AMF) crystallization was evaluated. The impact of esterification was also studied using three triglycerides. Melted blends containing the additives at concentrations lower than 12wt.% were quenched at 25°C and isothermal crystallization was monitored by pulsed low-resolution nuclear magnetic resonance. In parallel, polarized light microscopy was used to observe the microstructure. Compounds based on long chain saturated , i.e. , stearic, eicosanoic , tripalmitin and tristearin accelerated crystallization. Conversely, propanoic, hexanoic and oleic slowed down the process, while triacetin had no impact. Interestingly, above a critical concentration, the addition of , stearic or eicosanoic caused a transition from a one-step to two-step process. Gompertz model was used to fit the experimental data and to assess the influence of the molecular properties of the additives on the kinetic parameters.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Chemical compositions by using LC-MS/MS and GC-MS and biological activities of Sedum sediforme (Jacq.) Pau.

In this research, the chemical composition and biological activities of various extracts obtained from whole parts of Sedum sediforme (Jacq.) Pau were compared. The amounts of total phenolic and flavonoid components in crude extracts were determined by expression as pyrocatechol and quercetin equivalents, respectively. All of the extracts (petroleum ether, acetone, methanol, and water) obtained from S. sediforme showed strong antioxidant activity in four tested methods. Particularly, the IC50 values of the methanol extract, which was the richest in terms of total phenolic and flavonoid contents, were found to be lower than those of α-tocopherol and BHT in β-carotene bleaching (9.78 ± 0.06 μg/mL), DPPH free radical scavenging (9.07 ± 0.07 μg/mL), and ABTS cation radical scavenging (5.87 ± 0.03 μg/mL) methods. Furthermore, the methanol extract of S. sediforme showed higher inhibition activity than galanthamine against acetyl- and butyryl-cholinesterase enzymes. Also, acetone and methanol extracts exhibited moderate antimicrobial activity against Candida albicans. The main constituents of and essential oil were identified as (C16:0) (28.8%) and α-selinene (20.4%), respectively, by GC-MS. In the methanol extract of S. sediforme, quercetin, rutin, naringenin, and protocatechuic, p-coumaric, caffeic, and chlorogenic were detected and quantified by LC-MS/MS. Results of the current study showed that the methanol extract of S. sediforme may also be used as a food supplement.

Keyword: SCFA

Identification and evaluation of antibacterial agents present in lipophilic fractions isolated from sub-products of Phoenix dactilyfera.

In this study, the dried powder of seeds and pollen of Phoenix dactylifera was extracted using Soxhlet extractor and biochemical composition of lipophilic extracts was analysed by GCMS. Then, we have tested the antibacterial effect of fatty acids and volatile compounds found in these sub-products. The results revealed that the main fatty acids found in Deglet Nour seeds is are oleic (42.06%) followed by capric (24.16%), Takerbucht seeds extract contains mostly oleic (40.59%) and (20.19%), while Bent Kbala seed extract is rich in cis-vaccenic (52.09%) followed by capric (16.71%). Pollen extract have shown for the first time the richness in 14-methyl (33.31%), linoleic (31.16%) and linolenic (25.78%). Some of these elements seem to have a significant effect against some pathogenic bacteria.

Keyword: SCFA

Generation of the spiroketals conophthorin and chalcogran by fungal spores on polyunsaturated common to almonds and pistachios.

The spiroketal (E)-conophthorin has recently been reported as a semiochemical of the navel orangeworm moth, a major insect pest of California pistachios and almonds. Conophthorin and the isomeric spiroketal chalcogran are most commonly known as semiochemicals of several scolytid beetles. Conophthorin is both an insect- and plant-produced semiochemical widely recognized as a nonhost plant from the bark of several angiosperm species. Chalcogran is the principal aggregation pheromone component of the six-spined spruce bark beetle. Recent research has shown conophthorin is produced by almonds undergoing hull-split, and both spiroketals are produced by mechanically damaged almonds. To better understand the origin of these spiroketals, the emissions of orchard fungal spores on common to both pistachios and almonds were evaluated. The emission for the first 13 days of spores placed on a was monitored. The spores investigated were Aspergillus flavus (atoxigenic), A. flavus (toxigenic), Aspergillus niger, Aspergillus parasiticus, Penicillium glabrum, and Rhizopus stolonifer. The used as growth media were , oleic, linoleic, and linolenic. Spores on linoleic produced both spiroketals, those on linolenic produced only chalcogran, and those on and oleic did not produce either spiroketal. This is the first report of the spiroketals conophthorin and chalcogran from a fungal source.

Keyword: SCFA

[Studies on chemical constituents from roots of Peucedanum praeruptorum III].

To study the chemical constituents in roots of Peucedanum praeruptorum.The constituents were isolated by column chromatography on silica gel and ODS, and identified by spectroscopic methods.Seven compounds, alpha-D-glucopyranose-1-hexadecanoate (1), D-mannitol monohexadecanoate (2), adenosine (3), butyric (4), eleutheroside B, (5), apiosylskimmin (6), and mannitol (7) were isolated and identified.Compounds 1-5 were isolated from this plant for the first time.

Keyword: SCFA

Effect of γ-irradiation on bioactivity, compositions and compounds of clary sage seed (Salvia sclarea L.).

Clary sage seeds (Salvia sclarea L.) were obtained from plants cultivated, and 2.5, 4.0, 5.5, and 7.0 kGy doses of γ-irradiation were applied to the clary sage seeds. They were then analyzed for their protein, ash, oil and dry matter contents, and composition. Additionally, the total phenolic contents, antiradical, antioxidant activities, and compounds of the clary sage seed extract were determined. There was no significant difference in protein content. However, the moisture, oil, and ash contents of the samples were affected by irradiation. While the 7 kGy dose had a positive effect on the total phenolic content and antiradical activity of the sage seed extract, all doses have negative effects on the antioxidant activity of the sage seed. The main of the sage seed was remarkably found as α-linolenic . The four irradiation levels caused significant differences in composition by affecting all except , palmitoleic, and eicosenoic . The dominant compounds of control sage seed were found as β-pinene (18.81%) and limonene (15.60%). Higher doses of the irradiation decreased components of sage seed. Clary sage seed including high omega-3 can be irradiated with low doses (≤ 2.5 kGy) of γ-irradiation.Clary sage is one of the most popular Salvia species in Turkey and many countries. Clary sage seed has approximately 29% oil content and this oil contains >50% of α-linolenic . γ-Irradiation is widely applied in the preservation of spice quality. The present study shows that the antioxidant activity of the clary sage seed is decreased by γ-irradiation. Additionally, higher doses of irradiation also decreased the components of sage seed. Therefore, we suggest that clary sage seed which includes high levels of omega-3 should be irradiated with low doses (≤ 2.5 kGy) of γ-irradiation.© 2011 Institute of Food Technologists®

Keyword: SCFA

Identification and biosynthesis of novel male specific esters in the wings of the tropical butterfly, Bicyclus martius sanaos.

Representatives of the highly speciose tropical butterfly genus Bicyclus (Lepidoptera: Nymphalidae) are characterized by morphological differences in the male androconia, a set of scales and hair pencils located on the surface of the wings. These androconia are assumed to be associated with the release of courtship pheromones. In the present study, we report the identification and biosynthetic pathways of several novel esters from the wings of male B. martius sanaos. We found that the compounds in this male butterfly were similar to female-produced moth sex pheromones. Components associated with the male wing androconial areas were identified as ethyl, isobutyl and 2-phenylethyl hexadecanoates and (11Z)-11-hexadecenoates, among which the latter are novel natural products. By topical application of deuterium-labelled and amino precursors, we found these pheromone candidates to be produced in patches located on the forewings of the males. Deuterium labels from hexadecanoic were incorporated into (11Z)-11-hexadecenoic , providing experimental evidence of a Δ11-desaturase being active in butterflies. This unusual desaturase was found previously to be involved in the biosynthesis of female-produced sex pheromones of moths. In the male butterflies, both hexadecanoic and (11Z)-11-hexadecenoic were then enzymatically esterified to form the ethyl, isobutyl and 2-phenylethyl esters, incorporating ethanol, isobutanol, and 2-phenylethanol, derived from the corresponding amino L-alanine, L-valine, and L-phenylalanine.

Keyword: SCFA

Influence of acetic, citric, and lactic on Escherichia coli O157:H7 membrane lipid composition, verotoxin secretion, and resistance in simulated gastric fluid.

The effect of organic (acetic, citric, and lactic ) adaptation at equivalent initial pH values (6.4 and 5.4) on changes in membrane lipid composition, verotoxin concentration, and resistance in simulated gastric fluid (pH 1.5, 37 degrees C) was determined for Escherichia coli O157:H7 ATCC 43895 (HEC) and an rpoS mutant of E. coli O157:H7 ATCC 43895 (RM, FRIK 816-3). For HEC, lactic -adapted (pH 5.4) cells had the greatest D-value (32.2 min) and acetic -adapted (pH 5.4) cells had the smallest D-value (16.6 min) in simulated gastric fluid. For RM, D-values of citric and acetic -adapted cells were similar to those for nonadapted cells grown at pH 7.3, but D-values increased from 13.1 to 27.9 min in lactic -adapted cells (from pH 7.3 to pH 5.4). For both strains, the ratio of cis-vaccenic to decreased for citric and lactic -adapted cells, but the ratio increased for acetic -adapted cells at pH 5.4. Organic -adapted cells produced less total verotoxin than did nonadapted cells at approximately 10(8) CFU/ml. Extracellular verotoxin concentration proportionally decreased with decreasing pH for both HEC and RM. Changes in membrane lipid composition, verotoxin concentration, and resistance in HEC and RM were dependent on both pH and organic . Deletion of the rpoS gene did not affect these changes but did decrease resistance in citric -adapted cells. Results indicate that decreased membrane fluidity may have caused increased resistance and decreased verotoxin secretion.

Keyword: SCFA

Phytochemical investigation and antifeedant activity of Premna latifolia leaves.

The essential oil of Premna latifolia Roxb. was obtained by hydrodistillation of fresh leaves of the plant having an oil yield of 0.05%, both non-polar and essential oil were analysed by GC and GC-MS. Hexane fraction of the leaves of P. latifolia was transesterified and analysed by GC and GC-MS, 40 non-polar components were identified comprising 89.3%. The most abundant constituents were hexadecanoic (25.04%), 8,11,14-docosatrienoic (13.62%), octadecanoic (6.82%), 9,12-octadecadienoic (4.19%) and 29 components were investigated in the essential oil which comprises 78.1%. The most abundant oil constituents were 1-octen-3-ol (35.69%), terpendiol II (7.19%), δ-guaiene (7.49%) 2-undecanone (4.80%) and α-pinene (3.27%). Different extracts were also tested against polyphagous crop pest Spodoptera litura for antifeedant activity. Essential oil showed maximum growth reduction of 56.83% followed by chloroform extract of 43.93%.

Keyword: SCFA

Chemical profile by LC-MS/MS, GC/MS and antioxidant activities of the essential oils and crude extracts of two Euphorbia species.

In this study, it was aimed to investigate the chemical composition and antioxidant activities of two Euphorbia species. The major component of the compositions obtained from the petroleum ether extracts was identified as for Euphorbia gaillardotii and Euphorbia macroclada. The main constituents of the essential oils were identified as arachidic for E. gaillardotii and tetratetracontane for E. macroclada. Among the 27 studied compounds, hesperidin, rutin, hyperoside and quinic, malic, gallic and tannic were found to be the most abundant compounds in the two Euphorbia species. The methanol extracts of E. gaillardotii and E. macroclada showed strong antioxidant activity in all tested methods. Particularly, IC50 values of E. macroclada methanol extract that was the richest in terms of total phenolic-flavonoid contents were found to be lower than α-tocopherol and butylated hydroxytoluene in β-carotene bleaching, 2,2-diphenyl-1-picrylhydrazyl free and ABTS cation radical scavenging methods.

Keyword: SCFA

Characteristics of lipid extraction from Chlorella sp. cultivated in outdoor raceway ponds with mixture of ethyl acetate and ethanol for biodiesel production.

In this work, neutral lipids (NLs) extraction capacity and selectivity of six solvents were firstly compared. In addition, an eco-friendly solvent combination of ethyl acetate and ethanol (EA/E) was proposed and tested for lipid extraction from Chlorella sp. cultivated in outdoor raceway ponds and effect of extraction variables on lipid yield were intensively studied. Results indicated that lipid extraction yield was increased with solvent to biomass ratio but did not vary significantly when the value exceeded 20:1. Lipid yield was found to be strongly dependent on extraction temperature and time. Finally, profiles of lipid were determined and results indicated that the major components were octadecanoic , , linoleic and linolenic , demonstrating that the lipid extracted from the Chlorella sp. cultivated in outdoor raceway ponds by EA/E was suitable feedstock for biodiesel production.Copyright © 2015. Published by Elsevier Ltd.

Keyword: SCFA

Relation between salt tolerance and biochemical changes in cumin (Cuminum cyminum\xa0L.) seeds.

In this study, the effects of salinity on growth, , essential oil, and phenolic composition of cumin (Cuminum cyminum L.) seeds as well as the antioxidant activities of their extracts were investigated. Plants were treated with different concentrations of NaCl treatment: 0, 50, 75, and 125 mmoL. Plant growth was significantly reduced with the severity of saline treatment. This also caused important reductions in the seed yield and yield components. Besides, NaCl treatments affected composition. Petroselinic and linoleic proportions diminished consistently with the increase in NaCl concentration, whereas proportion increased. Furthermore, NaCl enhanced essential oil production in C. cyminum seeds and induced marked changes on the essential oil quality. Essential oil chemotype was modified from γ-terpinene/1-phenyl-1,2 ethanediol in control to γ-terpinene/β-pinene in salt stressed plants. Total polyphenol content was higher in treated seeds, and salinity improved the amount of individual phenolic compounds. Moreover, antioxidant activities of the extracts were determined by four different test systems, namely 2,2-diphenyl-1-picrylhydrazyl, β-carotene/linoleic chelating, and reducing power assays. The highest antioxidant activities were reveled in severe stressed plants. In this case, cumin seeds produced under saline conditions may function as a potential source of essential oil and antioxidant compounds, which could support the utilization of this plant in a large field of applications such as food industry.Copyright © 2016. Published by Elsevier B.V.

Keyword: SCFA

Profile of Herniaria fontanesii Growing Spontaneously in Tunisia.

The essential oil extracted from Desfontaine\'s rupturewort, Herniaria fontanesii J. Gay subsp. fontanesii growing wildly in Tunisia, was analyzed using GC and GC-MS techniques. The free radical scavenging capacity and total phenol contents of three crude extracts having different polarities (n-hexane, ethyl acetate and methanol) were examined. Thus, a total of 35 constituents were identified in the Desfontaine\'s rupturewort essential oil representing 89.8% of the whole constituents. The oil was dominated by hexadecanoic , caryophyllene oxide, terpin-4-ol, khusimone and trans-sabinene hydrate. The total phenolic contents ranged from 16.91 to 92.27 mg of gallic /g of dry weight and they were found to be significantly higher in methanol than in polar ethyl acetate and hexane extracts. Correlations were observed between the phenolic contents and the antioxidant properties. Thus, the antioxidant activity of the methanol extract was superior to that of all samples tested (IC50 = 0.21 ± 0.04 mg/mL).© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Keyword: SCFA

analogues exhibiting antinociceptive activity in mice.

Three derivatives were synthesized and evaluated as a potential platform for antinociceptive drug development. Female Swiss Webster mice were given N-(4-Methoxy-2-nitrophenyl)hexadecanamide (1), 2-amino-3-(palmitoylamino)benzoic (2) or 4-amino-3-(palmi-toylamino)benzoic (3) orally in doses of 10-100 mg/kg. The animals were tested for nociception using the hot plate and abdominal constriction response (writhing) tests. Compound 1 generated a dose-dependent antinociceptive effect, reflected by longer latencies (paw-lick and escape responses) and a decrease in writhing. Morphine (1.5-6 mg/kg, p.o.) and diclofenac (10-100 mg/kg, p.o.) were used as positive controls, respectively. Compounds 2 and 3 were less active in both nociceptive tests. The antinociception provoked by compound 1 was partially blocked by naloxone (1 mg/kg, i.p.) suggesting that this pharmacological effect could be due to the activation of micro-opioid receptors. N-(4-Methoxy-2-nitrophenyl)hexadecanamide showed antinociceptive effects in both nociceptive tests suggesting the possibility that this compound may define a new type of antinociceptive.

Keyword: SCFA

Metabolomic and transcriptomic responses induced in the livers of pigs by the long-term intake of resistant starch.

The present study investigated metabolomic and transcriptomic responses in the livers of pigs to evaluate the effects of resistant starch on the body\'s metabolism at the extraintestinal level. Thirty-six Duroc× Landrace × Large White growing barrows (70 d of age) were randomly allocated to either the corn starch (CS) group or the raw potato starch (RPS) group with a randomized complete block design; each group consisted of 6 replicates (pens), with 3 pigs per pen. Pigs in the CS group were offered a corn-soybean-based diet, whereas pigs in the RPS group were put on a diet in which 230 (growing) or 280 g/kg (finishing) purified CS was replaced with purified RPS during a 100-d trial. The livers of pigs were collected for metabolome and gene expression analysis. Gas chromatography-mass spectrometry analysis showed that compared with the CS diet, the RPS diet decreased ( < 0.05) cholesterol and as well as increased ( < 0.05) 3-hydroxybutyric , which indicated the reduction of adipose weight and biosynthesis and the elevation of β-oxidation. In addition, 2-ketoglutaric and glucose-6-phosphate were increased (< 0.05) although pyruvic was decreased ( < 0.05) in the RPS group, indicating the upregulated capacity of glucose phosphorylation and glycolysis. Microarray analysis showed that the mRNA expression of (), (), and () were downregulated ( < 0.05) whereas (), (), and () were upregulated ( < 0.05) in the RPS diet, indicating a decrease in intake and synthesis and an increase in oxidation and glycerophospholipid synthesis. The results demonstrated that the long-term consumption of RPS could modulate hepatic lipid metabolism by decreasing synthesis as well as increasing lipid oxidation and glycerophospholipid synthesis.

Keyword: SCFA

Chemical profile and biological activities of Veronica thymoides subsp. pseudocinerea.

In Turkey, Veronica species (Plantaginaceae) have been used as a diuretic and for wound healing in traditional medicine.To examine the and essential oil profiles, the antioxidant, anticholinesterase, antimicrobial, and DNA damage effects of Veronica thymoides P.H. Davis subsp. pseudocinerea M.A. Fischer as a potential source of natural active compounds.GC/MS was used to analyze essential oil and obtained from whole plant. The antioxidant activity was evaluated by the β-carotene-linoleic test system, DPPH-free and ABTS cation radicals scavenging, and cupric reducing antioxidant capacity assays. The anticholinesterase and antimicrobial activities were determined by Ellman and broth macrodillution methods, respectively. The effect of the methanol extract on DNA cleavage was investigated.Hexatriacontene (21.0%) was found to be the main constituent in essential oil, and linoleic (25.2%) and (20.6%) in . Methanol extract demonstrated the best IC50 values in lipid peroxidation (49.81 ± 0.31 µg/ml) and DPPH-free radical scavenging activity (15.32 ± 0.17 µg/ml). Methanol and water extracts possessed strong ABTS cation radical scavenging activity with IC50 values 9.15 ± 0.28 and 8.90 ± 0.1 \u2009µg/ml, respectively. The acetone extract exhibited moderate butyrylcholinesterase inhibitory activity. The highest antimicrobial activity was determined in methanol extract against Escherichia coli with 31.25 µg/ml MIC value. Inhibition of methanol extract on plasmid DNA cleavage by OH radicals was found to be 93.32% at 500 µg/ml.The methanol extract having strong antioxidant and DNA damage effects could be investigated phytochemically to find natural active compounds.

Keyword: SCFA

constituents of the aerial parts of white salsify (Tragopogon porrifolius L., Asteraceae).

Tragopogon porrifolius L. grows as a vegetable in southern Italy and all parts of it are edible. In the present study the components of the aerial part are described. Gas chromatography and gas chromatography-mass spectrometry analysis showed the presence of 38 components in all. On the whole, the fraction was constituted mainly by carbonylic compounds (24.6%), phenols (21.5%) and and esters (19.7%). The most abundant compounds were 4-vinyl guaiacol (19.0%), hexadecanoic (17.9%), hexahydrofarnesylacetone (15.8%) and hentriacontane (10.7%).

Keyword: SCFA

Volatile components of horsetail (Hippuris vulgaris L.) growing in central Italy.

Hippuris vulgaris, also known as horsetail or marestail, is a freshwater macrophyte occurring in lakes, rivers, ponds and marshes. According to \'The IUCN Red List of Threatened Species\', H. vulgaris is at a high risk of extinction in Italy in the medium-term future. In the present study, we analysed for the first time the volatile composition of H. vulgaris growing in central Italy. For the purpose, the essential oil was obtained by hydrodistillation and analysed by GC-MS. The chemical composition was dominated by aliphatic compounds such as fatty acids (26.0%), ketones (18.7%) and alkanes (11.4%), whereas terpenoids were poorer and mostly represented by diterpenes (7.4%). n-Hexadecanoic (25.5%), hexahydrofarnesyl acetone (17.5%) and trans-phytol (7.4%) were the major volatile constituents. These compounds are here proposed as chemotaxonomic markers of the species.

Keyword: SCFA

Valproate uncompetitively inhibits arachidonic acylation by rat acyl-CoA synthetase 4: relevance to valproate\'s efficacy against bipolar disorder.

The ability of chronic valproate (VPA) to reduce arachidonic (AA) turnover in brain phospholipids of unanesthetized rats has been ascribed to its inhibition of acyl-CoA synthetase (Acsl)-mediated activation of AA to AA-CoA. Our aim was to identify a rat Acsl isoenzyme that could be inhibited by VPA in vitro.Rat Acsl3-, Acsl6v1- and Acsl6v2-, and Acsl4-flag proteins were expressed in E. coli, and the ability of VPA to inhibit their activation of long-chain to acyl-CoA was estimated using Michaelis-Menten kinetics.VPA uncompetitively inhibited Acsl4-mediated conversion of AA and of docosahexaenoic (DHA) but not of to acyl-CoA, but did not affect AA conversion by Acsl3, Acsl6v1 or Acsl6v2. Acsl4-mediated conversion of AA to AA-CoA showed substrate inhibition and had a 10-times higher catalytic efficiency than did conversion of DHA to DHA-CoA. Butyrate, octanoate, or lithium did not inhibit AA activation by Acsl4.VPA\'s ability to inhibit Acsl4 activation of AA and of DHA to their respective acyl-CoAs, when related to the higher catalytic efficiency of AA than DHA conversion, may account for VPA\'s selective reduction of AA turnover in rat brain phospholipids, and contribute to VPA\'s efficacy against bipolar disorder.Copyright © 2010 Elsevier B.V. All rights reserved.

Keyword: SCFA

Effects of frozen storage and vacuum packaging on free and composition of Turkish Motal cheese.

Effects of vacuum packaging and frozen storage were studied on the formation of free (FFAs), compounds and microbial counts of Motal cheese samples stored for a period of 180 days. The FFA concentration of Motal cheese samples increased throughout the storage period of 180 days. However, the FFA contents of samples stored at -18 °C showed considerably lower values than those of the samples stored at 4 °C. (C16:0) and oleic (C18:1) were the most abundant FFAs in all the treatments. The compounds detected by headspace solid-phase microextraction (HS-SPME) profile of Motal cheese consisted of 16 esters, 10 , 6 ketones, 4 alcohols, 3 aldehydes, styrene, p-cresol and m-cresol. Results showed that storage at -18 °C can limit the excessive compound formation. Samples stored at 4°C with vacuum packaging showed comparatively high concentration of esters, ketones and alcohols. Samples stored without vacuum packaging at 4°C showed 2-nonanone as the most abundant compound toward the end of storage period. Storage at 4°C under vacuum packaging decreased the mold-yeast counts of samples. Frozen storage could be a suitable method for storing the Motal cheese.

Keyword: SCFA

Formulation of Granules for Site-Specific Delivery of an Antimicrobial Essential Oil to the Animal Intestinal Tract.

Owing to proliferation of antibiotic-resistant bacteria, the use of antibiotics for livestock growth promotion is banned in many countries and alternatives to in-feed antibiotics are needed. Cinnamon essential oil exhibits strong in vitro antibacterial activity; however, direct addition of essential oils to animal feed has limited practicality due to their high volatility, odor, fast decomposition, and poor availability in the lower intestines. To solve these problems, we formulated trans-cinnamaldehyde (CIN) with an adsorbent powder and via a melt-solidification technique. Core granules of an optimized composition contained up to 48% wt/wt CIN. The granules were then coated with an enteric polymer to impart site-specific release of CIN. CIN was mostly retained in simulated gastric fluid and released rapidly (>80% under 2 h) in simulated intestinal fluids. Rapid CIN autoxidation into cinnamic was inhibited by adding 1% vol/vol eugenol, which maintained CIN stability for at least 1 y. The granule formulation increased the antimicrobial activity of CIN against Escherichia coli K88 slightly with a minimum bactericidal concentration of 450 μg/mL for CIN in lauric -based granules compared with 550-600 μg/mL for -based granules and free CIN, respectively. These results encourage the potential use of encapsulated CIN for control of animal enteric pathogens by oral in-feed administration.Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Effects of on growth and poly-3-hydroxybutyrate production in bacteria.

The effects of saturated and unsaturated (lauric , , steric , oleic , linoleic , soybean oil) on Sphaerotilus natans, 0B17 (Pseudomonas sp.), and recombinant Escherichia coli DH5(/pUC19/CAB were studied. Oleic enhances Poly-3-hydroxybutyrate (PHB) production in these three bacterial strains, suggesting that the single double bond of the activates the polyhydroxylkanoate accumulation enzymatic reaction. Under the effect of lauric and linoleic , the growth of S. natans and 0B17 were totally inhibited. However, the enhanced PHB accumulation in recombinant E. coli was observed.

Keyword: SCFA

Enzymatic interesterification of palm stearin with Cinnamomum camphora seed oil to produce zero-trans medium-chain triacylglycerols-enriched plastic fat.

It is known that Cinnamomum camphora seed oil (CCSO) is rich in medium-chain (MCFAs) or medium-chain triacylglycerols (MCTs). The purpose of the present study was to produce zero-trans MCTs-enriched plastic fat from a lipid mixture (500 g) of palm stearin (PS) and CCSO at 3 weight ratios (PS:CCSO 60:40, 70:30, 80:20, wt/wt) by using lipase (Lipozyme TL IM, 10% of total substrate) as a catalyst at 65 °C for 8 h. The major of the products were (C16:0, 42.68% to 53.42%), oleic (C18:1, 22.41% to 23.46%), and MCFAs (8.67% to 18.73%). Alpha-tocopherol (0.48 to 2.51 mg/100 g), γ-tocopherol (1.70 to 3.88 mg/100 g), and δ-tocopherol (2.08 to 3.95 mg/100 g) were detected in the interesterified products. The physical properties including solid fat content (SFC), slip melting point (SMP), and crystal polymorphism of the products were evaluated for possible application in shortening or margarine. Results showed that the SFCs of interesterified products at 25 °C were 9% (60:40, PS:CCSO), 18.50% (70:30, PS:CCSO), and 29.2% (80:20, PS:CCSO), respectively. The β\' crystal form was found in most of the interesterified products. Furthermore, no trans were detected in the products. Such zero-trans MCT-enriched fats may have a potential functionality for shortenings and margarines which may become a new type of nutritional plastic fat for daily diet.© 2012 Institute of Food Technologists®

Keyword: SCFA

Effects of irrigation regimes on composition, antioxidant and antifungal properties of volatiles from fruits of Koroneiki cultivar grown under Tunisian conditions.

The olive tree is generally grown under rain-fed conditions. However, since the yield response to irrigation is great, even with low amounts of water, there is increasing interest in irrigated agriculture. The main goal of this study was, therefore, to investigate the effect of irrigation regimes on olive (Olea europaea L., cv. Koroneiki) obtained from an intensively-managed orchard in a semi-arid area with a Mediterranean climate in Tunisia. Different irrigation treatments 50% ETc, 75% ETc and 100% ETc were applied to the olive orchard. Accordingly, the effects of three irrigation regimes on compounds, composition and biological activities of Koroneiki cultivar were studied. The total profile of the constituents of all samples revealed the predominance of 3-ethenylpyridine (from 14.9-19.6%), phenylethyl alcool (from 7.8-19.2%) and benzaldehyde (from 9.0 to 13.8%). During watering level treatments studied, the major were oleic, and linoleic. Antioxidant activity of the fresh fruit volatiles cultivated at a watering level of 100% ETc was higher than that obtained under 50 and 75% Etc. The results of antifungal activity showed that the fruits volatiles of the three irrigation treatments had varying degrees of growth inhibition against the microorganisms tested.

Keyword: SCFA

Developing FT-NIR and PLS1 Methodology for Predicting Adulteration in Representative Varieties/Blends of Extra Virgin Olive Oils.

It was previously demonstrated that Fourier transform near infrared (FT-NIR) spectroscopy and partial least squares (PLS1) were successfully used to assess whether an olive oil was extra virgin, and if adulterated, with which type of vegetable oil and by how much using previously developed PLS1 calibration models. This last prediction required an initial set of four PLS1 calibration models that were based on gravimetrically prepared mixtures of a specific variety of extra virgin olive oil (EVOO) spiked with adulterants. The current study was undertaken after obtaining a range of EVOO varieties grown in different countries. It was found that all the different types of EVOO varieties investigated belonged to four distinct groups, and each required the development of additional sets of specific PLS1 calibration models to ensure that they can be used to predict low concentrations of vegetable oils high in linoleic, oleic, or , and/or refined olive oil. These four distinct sets of PLS1 calibration models were required to cover the range of EVOO varieties with a linoleic content from 1.3 to 15.5\xa0% of total . An FT-NIR library was established with 66 EVOO products obtained from California and Europe. The quality and/or purity of EVOO were assessed by determining the FT-NIR Index, a measure of the content of EVOO. The use of these PLS1 calibration models made it possible to predict the authenticity of EVOO and the identity and quantity of potential adulterant oils in minutes.

Keyword: SCFA

Zinc metalloproteinase ProA directly activates Legionella pneumophila PlaC glycerophospholipid:cholesterol acyltransferase.

Enzymes secreted by Legionella pneumophila, such as phospholipases A (PLAs) and glycerophospholipid:cholesterol acyltransferases (GCATs), may target host cell lipids and therefore contribute to the establishment of Legionnaires disease. L. pneumophila possesses three proteins, PlaA, PlaC, and PlaD, belonging to the GDSL family of lipases/acyltransferases. We have shown previously that PlaC is the major GCAT secreted by L. pneumophila and that the zinc metalloproteinase ProA is essential for GCAT activity. Here we characterized the mode of PlaC GCAT activation and determined that ProA directly processes PlaC. We further found that not only cholesterol but also ergosterol present in protozoa was palmitoylated by PlaC. Such ester formations were not induced by either PlaA or PlaD. PlaD was shown here to possess lysophospholipase A activity, and interestingly, all three GDSL enzymes transferred short chain to sterols. The three single putative catalytic amino (Ser-37, Asp-398, and His-401) proved essential for all PlaC-associated PLA, lysophospholipase A, and GCAT activities. A further four cysteine residues are important for the PLA/GCAT activities as well as their oxidized state, and we therefore conclude that PlaC likely forms at least one disulfide loop. Analysis of cleavage site and loop deletion mutants suggested that for GCAT activation deletion of several amino within the loop is necessary rather than cleavage at a single site. Our data therefore suggest a novel enzyme inhibition/activation mechanism where a disulfide loop inhibits PlaC GCAT activity until the protein is exported to the external space where it is ProA-activated.

Keyword: SCFA

Beneficial Effects of Co-Ultramicronized Palmitoylethanolamide/Luteolin in a Mouse Model of Autism and in a Case Report of Autism.

Autism spectrum disorder (ASD) is a condition defined by social communication deficits and repetitive restrictive behaviors. Association of the amide palmitoylethanolamide (PEA) with the flavonoid luteolin displays neuroprotective and antiinflammatory actions in different models of central nervous system pathologies. We hypothesized that association of PEA with luteolin might have therapeutic utility in ASD, and we employed a well-recognized autism animal model, namely sodium valproate administration, to evaluate cognitive and motor deficits.Two sets of experiments were conducted. In the first, we investigated the effect of association of ultramicronized PEA with luteolin, co-ultramicronized PEA-LUT® (co-ultraPEA-LUT®) in a murine model of autistic behaviors, while in the second, the effect of co-ultraPEA-LUT® in a patient affected by ASD was examined.Co-ultraPEA-LUT® treatment ameliorated social and nonsocial behaviors in valproic -induced autistic mice and improved clinical picture with reduction in stereotypes in a 10-year-old male child.These data suggest that ASD symptomatology may be improved by agents documented to control activation of mast cells and microglia. Co-ultraPEA-LUT® might be a valid and safe therapy for the symptoms of ASD alone or in combination with other used drugs.© 2016 John Wiley & Sons Ltd.

Keyword: SCFA

Nicotinic Receptor GPR109A Exerts Anti-Inflammatory Effects Through Inhibiting the Akt/mTOR Signaling Pathway in MIN6 Pancreatic β cells.

We found that activation of the nicotinic receptor GPR109A, expressed by the MIN6 murine pancreatic β cell line, inhibits nitric oxide accumulation induced by IFN-γ and TNF-α, implicating an anti-inflammatory effect of GPR109A in MIN6 cells. Nevertheless, the mechanism of its anti-inflammatory effect is still unknown. In this study, we used to stimulate MIN6 cells to induce inflammatory cytokine production and explored the mechanism by which GPR109A exerts anti-inflammatory effects.RT-PCR and immunocytochemical staining were used to detect the expression of GPR109A in MIN6 cells. Western blotting was used to detect the activation of the Akt/mTOR signaling pathway and expression of the inflammatory cytokine INF-γ, in MIN6 cells, following treatments with and +nicotinic , or with different concentrations of nicotinic and 3-hydroxybutyrate.In MIN6 cells, GPR109A transcripts and protein are expressed and GPR109A protein is mainly located in the cell membrane and cytoplasm. enhanced the phosphorylation of Akt and p70S6K and elevated the expression of IFN-γ. Co-treatment with nicotinic , which is an agonist of GPR109A, inhibited the -induced phosphorylation of Akt, mTOR, and p70S6K, as well as the expression of IFN-γ.GPR109A may inhibit inflammatory cytokine production, induced by , by MIN6 cells possibly via inhibiting the Akt/mTOR signaling pathway.© 2017 by the Association of Clinical Scientists, Inc.

Keyword: SCFA

Central composite design for the optimization of supercritical carbon dioxide fluid extraction of from Borago officinalis L. flower.

In the present study, and essential oils of the flower of borage (Borago officinalis\u2002L.) were obtained by supercritical carbon dioxide fluid extraction under different conditions. The extracts obtained were compared to oils of borage flower oil isolated by hydrodistillation. The obtained oils were analyzed by gas chromatography mass spectrometry. The compounds were identified according to their retention indices and mass spectra. The experimental parameters of supercritical fluid extraction (SFE) were optimized using a central composite design after a full factorial experimental design. Extraction yields based on SFE varied in the range of 0.02% to 1.96% (w/w), and the oil yield based on the hydrodistillation was 0.05% (v/w). The optimum conditions of SFE were obtained at a pressure of 350 atm, a temperature of 65 °C, a methanol modifier volume of 100 μL, and static and dynamic extraction time of 10 min. Main components of the extracts under optimum SFE conditions were , linoleic , γ-linolenic , and oleic . The results indicated that by using the suitable extraction conditions, SFE is more effective than the conventional hydrodistillation method in the extraction of and the preservation of its quality.SFE is a good technique for the extraction of oils from plants. The extraction yields by SFE are more than the conventional method. SFE is used on a large scale for production of essential oils and pharmaceutical products from plants.© 2011 Institute of Food Technologists®

Keyword: SCFA

Chemical composition and pharmacological significance of Anethum Sowa L. Root.

Medicinal herbs are used for the treatment of different ailments since antiquity. Different parts of Anethum sowa L. is used in folk medicine as a carminative for the treatment of flatulence, colic and hiccups of infants and children, antioxidant, antimicrobial and antispasmodic agent. The aim of our present study is to evaluate the chemical composition of the essential oil, proximate and elemental composition, amino , fatty profile and thermal behaviour of its root part as well as different pharmacological activities like antioxidant, antimicrobial and cytotoxicity of the root essential oil.The air-dried roots of Anethum sowa L. were subjected to hydro-distillation to yield the essential oil. The antioxidant activity of the essential oil was studied by DPPH radical scavenging activity. The antimicrobial activity was tested against four Gram-positive, six Gram-negative bacteria and four fungi species. The minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) for each examined microorganism were determined using the micro-dilution method. The LC value of the oil was also evaluated by brine shrimp lethality assay. The subsequent proximate analysis was also done by AOAC methods. The elemental analysis of the root powder was analysed by ICP-MS, AAS and FP system. The fatty was extracted by hot and cold extraction method and the analyses were carried out by GC. The amino profile was done by the amino analyzer. The DTA, DTG and TG of the root powder were taken by the thermogravimetric analyzer.A total of 24 constituents was identified and quantified in the essential oil and its water extract portion by GC and GC-MS. Apiol (81.99 and 74.779%) was found the highest phenylpropanoid constituent followed by m-diaminobenzene (10.446 and 8.778%) in the essential oil and aqueous extract portion. On the other hand, β-butyrolactone (5.13%) and isobutyl acetone (3.73%) were found in the major constituents in the water extract part. The IC value of the essential oil was found to be 3.07\xa0mg/mL by DPPH radical assay methods. The LC value of the brine shrimp cytotoxicity assay of the essential oil was observed at 0.81\xa0μg/mL. The essential oil showed better activity on Gram-negative bacteria than Gram-positive bacteria and fungi. The proximate composition showed that root contained 5.29% ash, 2.01% protein, 54.09% crude fibre, 0.15% essential oil and 1.14% fatty oil for hot extract and 0.23% for cold extract on the dried basis. The (33.81 & 31.58%) and linoleic (30.03 & 23.79%) were the major saturated and unsaturated fatty acids in the cold and hot extracted root powder respectively. Ca (23,600\xa0mg/kg), Mg (7620.33\xa0mg/kg) and K (1286.15\xa0mg/kg) were the most predominant elements followed by Ni (1187.30\xa0mg/kg), Se (913.79\xa0mg/kg), Li (317.84\xa0mg/kg), Na (288.72\xa0mg/kg) and Fe (206.88\xa0mg/kg). The toxic elements were found to be within the permissible limit. Glutamic (19.37%), glycine (14.53%) and lysine (17.08%) were found as the major amino acids. The decomposition rates were obtained by TG, DTG and DTA curve of the powder sample at various temperature ranges.The results demonstrated that the root part of Anethum sowa L. is a rich source of mineral elements, essential amino and fatty acids. The essential oil is the highly potential as bioactive oil for pharmaceuticals and medical applications, possessing antioxidant, antimicrobial and cytotoxic activities. The thermal analysis suggested as a simple, effective and rapid method to characterize the Anethum sowa L. species as well as to assess for herbal formulation.

Keyword: SCFA

Essential oil composition of lady\'s mantle (Alchemilla xanthochlora Rothm.) growing wild in Alpine pastures.

The fraction from the aerial part of Alchemilla xanthochlora Rothm. (Rosaceae) was obtained by hydrodistillation, and the chemical composition of the obtained oil was determined by GC/FID and GC/MS. Several compounds were found, belonging to the classes of aldehydes, alcohols, terpenes, esters, and hydrocarbons. The major constituents were cis-3-hexenol (11.20+/-0.02%), linalool (10.36+/-0.75%), oct-1-en-3-ol (8.98+/-1.43%), and nonanal (7.83+/-0.93%), followed by myrtenol (4.65+/-0.40%), hexadecanoic (4.59+/-0.79%), cis-3-hexenyl acetate (3.92+/-0.15%) and alpha-terpineol (3.10+/-0.26%).

Keyword: SCFA

Influence of Carotino oil on in vitro rumen fermentation, metabolism and apparent biohydrogenation of .

The study appraised the effects of Carotino oil on in vitro rumen fermentation, gas production, metabolism and apparent biohydrogenation of oleic, linoleic and linolenic . Carotino oil was added to a basal diet (50% concentrate and 50% oil palm frond) at the rate of 0, 2, 4, 6 and 8% dry matter of the diet. Rumen inoculum was obtained from three fistulated Boer bucks and incubated with 200\u2009mg of each treatment for 24\u2009h at 39°C. Gas production, fermentation kinetics, in vitro organic matter digestibility (IVOMD), (VFA), in vitro dry matter digestibility (IVDMD), metabolizable energy and free were determined. Carotino oil did not affect (P > 0.05) gas production, metabolizable energy, pH, IVOMD, IVDMD, methane, total and individual VFAs. However, Carotino oil decreased (P < 0.05) the biohydrogenation of linoleic and linolenic but enhanced (P < 0.05) the biohydrogenation of oleic . After 24\u2009h incubation, the concentrations of stearic, , pentadecanoic, myristic, myristoleic and lauric decreased (P < 0.05) while the concentration of linolenic, linoleic, oleic and transvaccenic and conjugated linoleic (CLAc9t11) increased (P < 0.05) with increasing levels of Carotino oil. Carotino oil seems to enhance the accumulation of beneficial unsaturated without disrupting rumen fermentation.© 2014 Japanese Society of Animal Science.

Keyword: SCFA

Does the fat tailed Damara ovine breed have a distinct lipid metabolism leading to a high concentration of branched chain in tissues?

Fat tailed sheep breeds are known for their adaptation to nutritional stress, among other harsh production conditions. Damara sheep, native to Southern Africa, have recently been exported to other areas of the world, particularly Australia, aiming to produce lamb in semi-arid regions. Damaras have a unique hanging fat tail, a fat depot able to be mobilized under nutritional stress. In this article we perform an in-depth characterization of the profiles of the fat tail in underfed and control Damara rams. Profiles were very similar between experimental groups, with the exception of (16:0) that was lower (P = 0.014) in underfed animals. However, the most striking result was the very high proportions of non-terminal branched chain found in the fat tail adipose tissue, as well as the gastrocnemius muscle of Damara rams. The muscle of Dorper and Merino rams used in the same experiment did not present non-terminal branched chain , suggesting that Damara rams have a unique lipid metabolism. Herein, we interpret this trait relating it to a higher ability of Damara sheep to digest fibrous fodder and to putative differences in the propionate metabolism by comparison to other sheep breeds.

Keyword: SCFA

[Metabonomic profiling of plasma metabolites in Wistar rats to study the effect of aging by means of GC/TOFMS-based techniques].

The global metabolite profiles of endogenous compounds of Wistar rats from 12 to 20 weeks old were investigated to take deep insight into and get better understanding of the pathogenesis of development and aging. Plasma from Wistar rats at 12, 14, 16, 18, and 20 weeks old were analyzed using GC/TOFMS. Multivariate data analysis was then used to process the metabonomic data which indicated excellent separation between different weeks and showed that the metabolic profiles of the samples changed with age, enabling age-related metabolic trajectories to be visualized. Decreased concentrations of citric , cis-aconitic , 9-(z)-hexadecenoic along with increased levels of hexanedioic , alpha-tocopherol, 3-indole propionic , etc contributed to the separation. Several major metabolic pathways were identified to be involved in metabolic regulation. This suggests that GC/TOFMS-based metabonomics is a powerful alternative approach to identifying potential biomarkers and investigating the physiological developments of aging and it is important to employ suitable age-match control group in metabonomic study of physiological monitoring, drug safety assessment, and disease diagnosis, etc.

Keyword: SCFA

Some physicochemical characteristics of pinus (Pinus halepensis Mill., Pinus pinea L., Pinus pinaster and Pinus canariensis) seeds from North Algeria, their lipid profiles and contents.

Physicochemical characteristics of seeds of some pinus species (Pinus halepensis Mill., Pinus pinea L., Pinus pinaster and Pinus canariensis) grown in North Algeria were determined. The results showed that the seeds consist of 19.8-36.7% oil, 14.25-26.62% protein, 7.8-8.6% moisture. Phosphorus, potassium and magnesium were the predominant elements present in seeds. Pinus seed\'s oil physicochemical properties show values (4.9-68.9), iodine values (93.3-160.4) and saponification values (65.9-117.9). Oil analysis showed that the major unsaturated for the four species were linoleic (30-59%) and oleic (17.4-34.6%), while the main saturated was (5-29%). Gas Chromatography and Mass Spectrometry analysis of P. halepensis Mill., P. pinaster and P. canariensis oils indicated that the major compound was the limonene with relative percentage of 3.1, 7.5 and 10.8, respectively.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Conserved valproic--induced lipid droplet formation in Dictyostelium and human hepatocytes identifies structurally active compounds.

Lipid droplet formation and subsequent steatosis (the abnormal retention of lipids within a cell) has been reported to contribute to hepatotoxicity and is an adverse effect of many pharmacological agents including the antiepileptic drug valproic (VPA). In this study, we have developed a simple model system (Dictyostelium discoideum) to investigate the effects of VPA and related compounds in lipid droplet formation. In mammalian hepatocytes, VPA increases lipid droplet accumulation over a 24-hour period, giving rise to liver cell damage, and we show a similar effect in Dictyostelium following 30 minutes of VPA treatment. Using (3)H-labelled polyunsaturated (arachidonic) or saturated () , we shown that VPA treatment of Dictyostelium gives rise to an increased accumulation of both types of in phosphatidylcholine, phosphatidylethanolamine and non-polar lipids in this time period, with a similar trend observed in human hepatocytes (Huh7 cells) labelled with [(3)H]arachidonic . In addition, pharmacological inhibition of β-oxidation in Dictyostelium phenocopies accumulation, in agreement with data reported in mammalian systems. Using Dictyostelium, we then screened a range of VPA-related compounds to identify those with high and low lipid-accumulation potential, and validated these activities for effects on lipid droplet formation by using human hepatocytes. Structure-activity relationships for these VPA-related compounds suggest that lipid accumulation is independent of VPA-catalysed teratogenicity and inositol depletion. These results suggest that Dictyostelium could provide both a novel model system for the analysis of lipid droplet formation in human hepatocytes and a rapid method for identifying VPA-related compounds that show liver toxicology.

Keyword: SCFA

[Lipid synthesis by an acidic tolerant Rhodotorula glutinis].

Acetic , as a main by-product generated in the pretreatment process of lignocellulose hydrolysis, significantly affects cell growth and lipid synthesis of oleaginous microorganisms. Therefore, we studied the tolerance of Rhodotorula glutinis to acetic and its lipid synthesis from substrate containing acetic . In the mixed sugar medium containing 6 g/L glucose and 44 g/L xylose, and supplemented with acetic , the cell growth was not:inhibited when the acetic concentration was below 10 g/L. Compared with the control, the biomass, lipid concentration and lipid content of R. glutinis increased 21.5%, 171% and 122% respectively when acetic concentration was 10 g/L. Furthermore, R. glutinis could accumulate lipid with acetate as the sole carbon source. Lipid concentration and lipid yield reached 3.20 g/L and 13% respectively with the initial acetic concentration of 25 g/L. The lipid composition was analyzed by gas chromatograph. The main composition of lipid produced with acetic was , stearic , oleic , linoleic and linolenic , including 40.9% saturated and 59.1% unsaturated . The lipid composition was similar to that of plant oil, indicating that lipid from oleaginous yeast R. glutinis had potential as the feedstock of biodiesel production. These results demonstrated that a certain concentration of acetic need not to be removed in the detoxification process when using lignocelluloses hydrolysate to produce microbial lipid by R. glutinis.

Keyword: SCFA

Oral bioavailability of the ether lipid plasmalogen precursor, PPI-1011, in the rabbit: a new therapeutic strategy for Alzheimer\'s disease.

Docosahexaenoic (DHA) and DHA-containing ethanolamine plasmalogens (PlsEtn) are decreased in the brain, liver and the circulation in Alzheimer\'s disease. Decreased supply of plasmalogen precursors to the brain by the liver, as a result of peroxisomal deficits is a process that probably starts early in the AD disease process. To overcome this metabolic compromise, we have designed an orally bioavailable DHA-containing ether lipid precursor of plasmalogens. PPI-1011 is an alkyl-diacyl plasmalogen precursor with at sn-1, DHA at sn-2 and lipoic at sn-3. This study outlines the oral pharmacokinetics of this precursor and its conversion to PlsEtn and phosphatidylethanolamines (PtdEtn).Rabbits were dosed orally with PPI-1011 in hard gelatin capsules for time-course and dose response studies. Incorporation into PlsEtn and PtdEtn was monitored by LC-MS/MS. Metabolism of released lipoic was monitored by GC-MS. To monitor the metabolic fate of different components of PPI-1011, we labeled the sn-1 , sn-2 DHA and glycerol backbone with (13)C and monitored their metabolic fates by LC-MS/MS.PPI-1011 was not detected in plasma suggesting rapid release of sn-3 lipoic via gut lipases. This conclusion was supported by peak levels of lipoic metabolites in the plasma 3 hours after dosing. While PPI-1011 did not gain access to the plasma, it increased circulating levels of DHA-containing PlsEtn and PtdEtn. Labeling experiments demonstrated that the PtdEtn increases resulted from increased availability of DHA released via remodeling at sn-2 of phospholipids derived from PPI-1011. This release of DHA peaked at 6 hrs while increases in phospholipids peaked at 12 hr. Increases in circulating PlsEtn were more complex. Labeling experiments demonstrated that increases in the target PlsEtn, 16:0/22:6, consisted of 2 pools. In one pool, the intact precursor received a sn-3 phosphoethanolamine group and desaturation at sn-1 to generate the target plasmalogen. The second pool, like the PtdEtn, resulted from increased availability of DHA released during remodeling of sn-2. In the case of sn-1 18:0 and 18:1 plasmalogens with [(13)C(3)]DHA at sn-2, labeling was the result of increased availability of [(13)C(3)]DHA from lipid remodeling. Isotope and repeated dosing (2 weeks) experiments also demonstrated that plasmalogens and/or plasmalogen precursors derived from PPI-1011 are able to cross both the blood-retinal and blood-brain barriers.Our data demonstrate that PPI-1011, an ether lipid precursor of plasmalogens is orally bioavailable in the rabbit, augmenting the circulating levels of unesterified DHA and DHA-containing PlsEtn and PtdEtn. Other ethanolamine plasmalogens were generated from the precursor via lipid remodeling (de-acylation/re-acylation reactions at sn-2) and phosphatidylethanolamines were generated via de-alkylation/re-acylation reactions at sn-1. Repeated oral dosing for 2 weeks with PPI-1011 resulted in dose-dependent increases in circulating DHA and DHA-containing plasmalogens. These products and/or precursors were also able to cross the blood-retinal and blood-brain barriers.

Keyword: SCFA

Suppression of monosodium urate crystal-induced cytokine production by butyrate is mediated by the inhibition of class I histone deacetylases.

Acute gouty arthritis is caused by endogenously formed monosodium urate (MSU) crystals, which are potent activators of the NLRP3 inflammasome. However, to induce the release of active interleukin (IL)-1β, an additional stimulus is needed. Saturated long-chain free (FFAs) can provide such a signal and stimulate transcription of pro-IL-1β. In contrast, the short-chain butyrate possesses anti-inflammatory effects. One of the mechanisms involved is inhibition of histone deacetylases (HDACs). Here, we explored the effects of butyrate on MSU+FFA-induced cytokine production and its inhibition of specific HDACs.Freshly isolated peripheral blood mononuclear cells (PBMCs) from healthy donors were stimulated with MSU and (C16.0) in the presence or absence of butyrate or a synthetic HDAC inhibitor. Cytokine responses were measured with ELISA and quantitative PCR. HDAC activity was measured with fluorimetric assays.Butyrate decreased C16.0+MSU-induced production of IL-1β, IL-6, IL-8 and IL-1β mRNA in PBMCs from healthy donors. Similar results were obtained in PBMCs isolated from patients with gout. Butyrate specifically inhibited class I HDACs. The HDAC inhibitor, panobinostat and the potent HDAC inhibitor, ITF-B, also decreased ex vivo C16.0+MSU-induced IL-1β production.In agreement with the reported low inhibitory potency of butyrate, a high concentration was needed for cytokine suppression, whereas synthetic HDAC inhibitors showed potent anti-inflammatory effects at nanomolar concentrations. These novel HDAC inhibitors could be effective in the treatment of acute gout. Moreover, the use of specific HDAC inhibitors could even improve the efficacy and reduce any potential adverse effects.Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

Keyword: SCFA

Effect of on the β-oxidation system and thioesterase of Lactococcus lactis subspecies lactis.

The influence of on the β-oxidation system and thioesterase of Lactococcus lactis was investigated in this study. The results showed that (C8:0-C16:0) significantly inhibited the growth of Lactococcus lactis, and laurate (C12:0) had the highest bactericidal effects. We detected the maximum activity of β-oxidation at different incubation times (8, 12, and 18h) to be 6.460, 7.751, and 8.203, respectively, and the maximum activity of thioesterase at different incubation times (8, 12, and 18h) to be 19.498, 27.180, and 12.800, respectively. were seen to induce the β-oxidation system and activity of thioesterase; decanoic (C10:0) and (C16:0) were also seen to induce the β-oxidation system of Lactococcus lactis, but the induced ability was significantly different. Octanoic (C8:0) and (C16:0) were seen to induce thioesterase activity in Lactococcus lactis. When 1mM (C16:0) was added to M17 broth, the activity of thioesterase increased 5-fold after 2min; however, adding octanoic (C8:0) changed the activity little. Evidence showed that the ability to induce the β-oxidation system and thioesterase activity was related to the \' chain lengths.Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Increasing the sensitivity of an LC-MS method for screening material extracts for organic extractables via mobile phase optimization.

Organic extractables (substances extracted from materials used in pharmaceutical packaging) are discovered, identified, and quantified via screening of extracts with analytical methods including liquid chromatography with mass spectrometric detection (LC-MS). Because extractables include a large number of diverse compounds that are typically present in plastic extracts at low levels, the LC-MS methods must be broad scope and sensitive. To accomplish these objectives, screening studies typically couple gradient reversed-phase separations with electrospray MS detection (both positive and negative ion modes). While such methods are generally applicable for a number of extractables, they are not optimal for some commonly encountered extractables due to either poor chromatographic performance (e.g., peak tailing) or poor MS response. Modifications to mobile phase composition (e.g., pH adjustment) were examined to improve the performance of an LC-MS screening method. The use of 0.1% acetic with 1 mM ammonium acetate (pH 3.6) as the aqueous portion of the mobile phase provided favorable sensitivities for a number of extractables both in positive and negative ion modes. In positive ion mode, the acidic mobile phase improved responses for moderately weak basic compounds by increasing their degree of protonation. For very weak basic compounds such as amides, ammonium ions in the mobile phase promoted proton adduct responses. In negative ion mode, an acidic mobile phase containing acetate anion improved ESI responses for acidic compounds, primarily due to gas phase effects.

Keyword: SCFA

Acetoacetate as regulator of -induced uncoupling involving liver mitochondrial ADP/ATP antiporter and aspartate/glutamate antiporter.

The effect of acetoacetate on palmitate-induced uncoupling with the involvement of ADP/ATP antiporter and aspartate/glutamate antiporter has been studied in liver mitochondria. The incubation of mitochondria with acetoacetate during succinate oxidation in the presence of rotenone, oligomycin, and EGTA suppresses the accumulation of conjugated dienes. This is considered as a display of antioxidant effect of acetoacetate. Under these conditions, acetoacetate does not influence the respiration of mitochondria in the absence or presence of palmitate but eliminates the ability of carboxyatractylate or aspartate separately to suppress the uncoupling effect of this . The action of acetoacetate is eliminated by beta-hydroxybutyrate or thiourea, but not by the antioxidant Trolox. In the absence of acetoacetate, the palmitate-induced uncoupling is limited by a stage sensitive to carboxyatractylate (ADP/ATP antiporter) or aspartate (aspartate/glutamate antiporter); in its presence, it is limited by a stage insensitive to the effect of these agents. In the presence of Trolox, ADP suppresses the uncoupling action of palmitate to the same degree as carboxyatractylate. Under these conditions, acetoacetate eliminates the recoupling effects of ADP and aspartate, including their joint action. This effect of acetoacetate is eliminated by beta-hydroxybutyrate or thiourea. It is supposed that the stimulating effect of acetoacetate is caused both by increase in the rate of transfer of anion from the inner monolayer of the membrane to the outer one, which involves the ADP/ATP antiporter and aspartate/glutamate antiporter, and by elimination of the ability of ADP to inhibit this transport. Under conditions of excessive production of reactive oxygen species in mitochondria at a high membrane potential and in the presence of small amounts of , such effect of acetoacetate can be considered as one of the mechanisms of antioxidant protection.

Keyword: SCFA

Comparative researches on two direct transmethylation without prior extraction methods for analysis in vegetal matrix with low fat content.

The aim of our work was to compare two methods, both based on direct transmethylation with different reagents, BF3/MeOH (boron trifluoride in methanol) or HCl/MeOH (hydrochloride in methanol), in catalysis, without prior extraction, to find the fast, non-expensive but enough precise method for 9 principal (lauric, myristic, , stearic, oleic, linoleic, linolenic, arahidic and behenic ) analysis in vegetal matrix with low fat content (forage from grassland), for nutrition and agrochemical studies.Comparatively, between the average values obtained for all analysed by the two methods based on direct transmethylation without prior extraction no significantly difference was identified (p > 0.05). The results of for the same forage sample were more closely to their average value, being more homogenous for BF3/MeOH than HCl/MeOH, because of the better accuracy and repeatability of this method. Method that uses BF3/MeOH reagent produces small amounts of interfering compounds than the method using HCl/MeOH reagent, results reflected by the better statistical parameters.The fast and non-expensive BF3/methanol method was applied with good accuracy and sensitivity for the determination of free or combined (saturated and unsaturated) in forage matrix with low fat content from grassland. Also, the final extract obtained by this method, poorer in interfering compounds, is safer to protect the injector and column from contamination with heavy or non- compounds formed by transmethylation reactions.

Keyword: SCFA

[Changes in amino and contents as well as activity of some related enzymes in apple fruit during aroma production].

Aroma volatiles from apple (Malus domestica Borkh. var. Starkrimson) fruit at different stages of maturity were collected by solid adsorbent-Tenax-GC and determined by thermodesorption and GC-MS. Production of propyl acetate, butyl acetate, ethyl 2-methyl-butanoate and total ester volatiles and changes in concentration of the precursors of aroma biosynthsis--free amino and and activities of lipoxygenases (LOX) and alcohol acetyltransferase (AAT) in apple fruits during ripening were studied. The results showed that propyl acetate and total esters were very low when the endogenous ethylene formation of the fruit was very low. At the stage of the increase in ethylene production, the rate of formation of propyl acetate and total esters increased. Butyl acetate appeared at the beginning of ethylene rise and increased thereafter. Ethyl 2-methyl-butanoate was produced at the beginning of climacteric stage and then increased sharply (Figs.1). These facts suggest that the aroma production is closely related to ethylene production. Among the 14 free amino detected in fruit, isoleucine which is considered to be the biosynthetic precursor of some branched chain esters showed a great increase during fruit ripening while the others decreased or remained stable (Table 1). The accumulation of isoleucine suggested that isoleucine supply in fruit may not limit the biosynthesis of esters with branched chain alkyl groups. Concentrations of free such as , linolenic, oleic, linoleic, stearic increased before the increase of aroma production, decreased with the increase of aroma production and showed an increase at postclimacteric stages (Fig.2). LOX activity increased at climacteric stages and declined rapidly thereafter. AAT activity increased sharply at the early stage of fruit maturity when the aroma was very low and remained at a stable high level during fruit ripening (Fig.3) indicating that the AAT activity is not the limiting factor for aroma formation in apple fruit.

Keyword: SCFA

Nrf2 affects the efficiency of mitochondrial oxidation.

Transcription factor Nrf2 (NF-E2 p45-related factor 2) regulates the cellular redox homoeostasis and cytoprotective responses, allowing adaptation and survival under conditions of stress. The significance of Nrf2\xa0in intermediary metabolism is also beginning to be recognized. Thus this transcription factor negatively affects synthesis. However, the effect of Nrf2 on oxidation is currently unknown. In the present paper, we report that the mitochondrial oxidation of long-chain () and short-chain (hexanoic) is depressed in the absence of Nrf2 and accelerated when Nrf2 is constitutively active. Addition of stimulates respiration in heart and liver mitochondria isolated from wild-type mice. This effect is significantly weaker when Nrf2 is deleted, whereas it is stronger when Nrf2 activity is constitutively high. In the absence of glucose, addition of differentially affects the production of ATP in mouse embryonic fibroblasts from wild-type, Nrf2-knockout and Keap1 (Kelch-like ECH-associated protein 1)-knockout mice. In acute tissue slices, the rate of regeneration of FADH2 is reduced when Nrf2 is absent. This metabolic role of Nrf2 on oxidation has implications for chronic disease conditions including cancer, metabolic syndrome and neurodegeneration.

Keyword: SCFA

Effect of synthetic ligands of PPAR α, β/δ, γ, RAR, RXR and LXR on the composition of phospholipids in mice.

Nuclear hormone receptors are transcription factors that can be activated by nutrition-derived ligands and alter the expression of various specific target genes. Stearoyl-Coenzyme A desaturase (SCD1) converts (16:0) to palmitoleic (16:1n-7) as well as stearic (18:0) to oleic (18:1n-9). At the same time, elongase 6 (ELOVL6) elongates 16:1n-7 and 18:1n-9 to vaccenic (18:1n-7) and eicosenoic (20:1n-9). We examined how synthetic selective ligands of nuclear hormone receptors alter the gene expression of hepatic enzymes in mice. In addition, we examined how the regulation of these two enzymes influences composition of phospholipids in liver and plasma. Mice were gavaged daily for 1 week with synthetic ligands of peroxisome proliferator-activated receptor (PPAR) α, β/δ, γ, liver X receptor (LXR), retinoic receptor (RAR) and retinoid-X receptor (RXR) for 1 week. Phospholipids from liver and plasma were analysed using ESI-MS/MS and GC after saponification. Hepatic gene expression of SCD1 and ELOVL6 was measured using QRT-PCR. SCD1 and ELOVL6 expression increased after the gavage of LXR and RXR ligands. The analysis of composition of total phospholipids in plasma and liver showed increased percentage contributions of the SCD1 and ELOVL6 products 18:1n-9, 18:1n-7 and 20:1n-9 after LXR and RXR ligand application. Analysis of total phospholipids from plasma and liver revealed a significant increase in monounsaturated bound in phosphatidylcholine (PtdCho) and lysophosphatidylcholine (PtdEtn) after LXR and RXR ligand administration. Increased hepatic gene expression of SCD1 and ELOVL6 after gavage of selective RXR or LXR ligands to mice resulted in increased concentrations of their metabolic products in phospholipids of liver and plasma.

Keyword: SCFA

Assessment of myocardial metabolism in diabetic rats using small-animal PET: a feasibility study.

This feasibility study was undertaken to determine whether kinetic modeling in conjunction with small-animal PET could noninvasively quantify alterations in myocardial perfusion and substrate metabolism in rats.All small-animal PET was performed on either of 2 tomographs. Myocardial blood flow and substrate metabolism were measured in 10 male Zucker diabetic rats (ZDF, fa/fa) and 10 lean littermates (Lean, Fa/+) using (15)O-water, 1-(11)C-glucose, 1-(11)C-acetate, and 1-(11)C-palmitate. Animals were 12.0 +/- 1.4-wk old.Consistent with a type 2 diabetic phenotype, the ZDF animals showed higher plasma hemoglobin A(1c), insulin, glucose, and free (FFA) levels than their lean controls. Myocardial glucose uptake (mL/g/min) was not significantly different between the 2 groups. However, higher glucose plasma levels in the ZDF rats resulted in higher myocardial glucose utilization (nmol/g/min) (Lean, 629 +/- 785, vs. ZDF, 1,737 +/- 1,406; P = 0.06). Similarly, myocardial FFA uptake (mL/g/min) was not significantly different between the 2 groups, (Lean, 0.51 +/- 28, vs. ZDF, 0.72 +/- 0.19; P = not significant) However, due to higher FFA plasma levels, utilization and oxidation (nmol/g/min) were significantly higher in the ZDF group (Lean, 519 +/- 462, vs. ZDF, 1,623 +/- 712, P < .001; and Lean, 453 +/- 478, vs. ZDF, 1,636 +/- 730, P < .01).Noninvasive measurements of myocardial substrate metabolism in ZDF rats using small-animal PET are consistent with the expected early metabolic abnormalities that occur in this well-characterized model of type 2 diabetes mellitus. Thus, small-animal PET demonstrates significant promise in providing a means to link the myocardial metabolic abnormalities that occur in rat of disease with the human condition.

Keyword: SCFA

Influence of pig rennet on composition, molecule profile, texture and sensory properties of Pecorino di Farindola cheese.

Pig rennet is traditionally used in Pecorino di Farindola cheese. In this study, different Pecorino cheeses obtained using calf, kid and pig rennets were compared in terms of molecule profile, texture and sensory properties during ripening.The rennet type influenced the composition of cheeses, though , myristic and oleic were always predominant. The analysis of volatiles by SPME-GC/MS showed that Pecorino from calf rennet, at the end of ripening, was the least \'evolved\' in terms of profile. SPME-GC/MS analysis revealed that cheeses from calf rennet showed the slowest accumulation of free over ripening time. data permitted the differentiation of cheese samples ripened from 30 to 180 days according to the rennet used. Texture analysis differentiated cheeses made with pig and calf rennet from those made with kid rennet, which were less hard and more elastic than the former. Also sensory analysis differentiated cheese samples on the basis of rennet type, and cheeses made with pig rennet showed the lowest elasticity, bitter taste and fruity and hay flavour intensities.Pig rennet is fundamental to determine the quality parameters of Pecorino di Farindola cheese and could be used to impart peculiar quality features to ewe\'s milk cheeses.© 2014 Society of Chemical Industry.

Keyword: SCFA

Antiproliferative effect of methanolic extraction of tualang honey on human keloid fibroblasts.

Keloid is a type of scar which extends beyond the boundaries of the original wound. It can spread to the surrounding skin by invasion. The use of Tualang honey is a possible approach for keloid treatment. The objective of this study was to determine the antiproliferative effect of methanolic extraction of Tualang honey to primary human keloid fibroblasts and to identify the compounds in methanol extraction of Tualang honey.Crude Tualang honey was extracted with methanol and then dried using rota vapor to remove remaining methanol from honey. Normal and keloid fibroblasts were verified and treated with the extracted honey. Cell proliferation was tested with [3-(4,5-dimethylthiazol-2-yi)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) assay. Extraction of Tualang honey using methanol was carried out and the extracted samples were analysed using gas chromatography-mass spectrometry (GC-MS). The result was analysed using SPSS and tested with Kruskal-Wallis and Mann-Whitney tests.Methanolic extraction of honey has positive anti proliferative effect on keloid fibroblasts in a dose-dependent manner. The presence of such as , stearic , oleic , linoleic and octadecanoic may contribute to the anti-proliferative effect in keloid fibroblasts.The methanolic honey extraction has an antiproliferative effect on keloid fibroblasts and a range of compounds has been identified from Tualang honey. The antiproliferative effect of keloid fibroblasts towards Tualang honey may involve cell signaling pathway. Identifying other compounds from different organic solvents should be carried out in future.

Keyword: SCFA

Identification of Predominant Phytochemical Compounds and Cytotoxic Activity of Wild Olive Leaves (Olea europaea L. ssp. sylvestris) Harvested in South Portugal.

This study has been aimed at providing a qualitative and quantitative evaluation of selected phytochemicals such as phenolic acids, flavonoids, oleuropein, fatty acids profile, and volatile oil compounds, present in wild olive leaves harvested in Portugal, as well as at determining their antioxidant and cytotoxic potential against human melanoma HTB-140 and WM793, prostate cancer DU-145 and PC-3, hepatocellular carcinoma Hep G2 cell lines, as well as normal human skin fibroblasts BJ and prostate epithelial cells PNT2. Gallic, protocatechuic, p-hydroxybenzoic, vanillic acids, apigenin 7-O-glucoside, luteolin 7-O-glucoside, and rutin were identified in olive leaves. The amount of oleuropein was equal to 22.64 g/kg dry weight. (E)-Anethole (32.35%), fenchone (11.89%), and (Z)-3-nonen-1-ol (8%) were found to be the main constituents of the oil volatile fraction, whereas , oleic, and alpha-linolenic were determined to be dominating fatty acids. Olive leaves methanol extract was observed to exerted a significant, selective cytotoxic effect on DU-145 and PC-3 cell lines. Except the essential oil composition, evaluated wild olive leaves, with regard to their quantitative and qualitative composition, do not substantially differ from the leaves of other cultivars grown for industrial purposes and they reveal considerable antioxidant and cytotoxic properties. Thus, the wild species may prove to be suitable for use in traditional medicine as cancer chemoprevention.© 2017 Wiley-VHCA AG, Zurich, Switzerland.

Keyword: SCFA

Pharmacological activities of the organic extracts and fatty composition of the petroleum ether extract from Haplophyllum tuberculatum leaves.

Haplophyllum tuberculatum is used in traditional medicine to treat many disorders including inflammation and pain. The aim of this study is to investigate the organic extracts from H. tuberculatum leaves against inflammation, gastric ulcer and pain.Acute toxicity was studied in vivo to determine the toxic doses of the organic extracts. Anti-inflammatory activity was also evaluated in vivo using carrageenan-induced paw edema in Wistar rats. Gastroprotective activity was tested using the HCl/ethanol-induced gastric ulcer test in rats. Peripheral and central analgesic activities were assessed using the acetic -induced writhing test and the hot-plate method, respectively. The chemical composition of the fatty acids in the petroleum ether (PE) extract was determined with GC-MS.At 25, 50 and 100mg/kg PE extract was the most active against inflammation. Percentages inhibition 5h after carrageenan-injection were 51.12; 86.71% and 96.92%, respectively. The same extract at 100mg/kg showed good analgesic activities using the acetic -induced writhing test and the hot-plate method. The chloroform, ethyl acetate (EtOAc) and butanolic (n-BuOH) extracts exhibited strong anti-inflammatory, gastroprotective and analgesic activities at 100mg/kg. The GC-FID analysis revealed that the PE extract was rich in γ-linolenic (45.50%) followed by (18.48%), linoleic (10.73%), erucic (4.72), stearic (3.96%) and oleic (2.57%).The results of the present study support the traditional use of the leaves of H. tuberculatum and may possibly serve as prospective material for further development of safe new phytochemical anti-inflammatory, gastroprotective and/or analgesic agents.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: SCFA

Greater dietary fat oxidation in obese compared with lean men: an adaptive mechanism to prevent liver fat accumulation?

Liver fat represents a balance between input, secretion, and oxidation of . As humans spend the majority of a 24-h period in a postprandial state, dietary make an important contribution to liver fat metabolism. We compared hepatic partitioning in healthy lean (n = 9) and abdominally obese (n = 10) males over 24 h. Volunteers received three mixed meals adjusted for basal metabolic rate. U-13C-labeled were incorporated into the meals, and [2H2]palmitate was infused intravenously to distinguish between sources of incorporated into VLDL-TG. Immunoaffinity chromatography was used to isolate VLDL-TG of hepatic origin. Liver and whole body oxidation was assessed by isotopic enrichment of 3-hydoxybutyrate and breath CO2. We found a similar contribution of dietary to VLDL-TG in the two groups over 24 h. The contribution of from splanchnic sources was higher (P < 0.05) in the abdominally obese group. Ketogenesis occurred to a significantly greater extent in abdominally obese compared with lean males, largely due to lessened downregulation of postprandial ketogenesis (P < 0.001). The appearance of 13C in breath CO2 was also greater (P < 0.001) in abdominally obese compared with lean men. Hepatic elongation and desaturation of were higher (P < 0.05) in abdominally obese than in lean males. Oxidation of dietary and hepatic desaturation and elongation of occurred to a greater extent in abdominally obese men. These alterations may represent further pathways for redirection of into export from the liver or oxidation to prevent liver fat accumulation.

Keyword: SCFA

Antibacterial activity of extracted bioactive molecules of Schinus terebinthifolius ripened fruits against some pathogenic bacteria.

The aim of this work is to identify the chemical constituents and the bioactivity of essential oil (EO), acetone extract (ACE) and n-hexane extract (HexE) of S. terebinthifolius ripened fruits using GC-MS. Total phenolic content and antioxidant activity of extracts were determined using the Folin-Ciocalteu and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assays, respectively. The toxicity against the growth of Acinetobacter baumannii, Bacillus subtilis, Escherichia coli, Micrococcus flavus, Pseudomonas aeruginosa, Sarcina lutea, and Staphylococcus aureus was determined with measuring the inhibition zones (IZs) using the disc diffusion method at the concentrations from 125 to 2000\u202fμg/mL, also, the minimum inhibitory concentrations (MICs) using 96-well micro-plates and ranged from 4 to 2000\u202fμg/mL. The major components in EO were α-pinene (36.9%), and α-phellandrene (32.8%). The major components in ACE were oleic (38.7%), α-phellandrene (13.33%), and δ-cadinene (11.1%), while the major methyl esters of detected in HexE were oleic (12.8%), and (10.9%). The EO showed good activity against the growth of Staph. aureus and P. aeruginosa with MIC values of 16\u202fμg/mL and 32\u202fμg/mL, the ACE showed broad activity against the studied bacterial pathogens with MIC values ranged from of 4-128\u202fμg/mL against the studied bacterial isolates, while HexE, however, showed weak antibacterial activity. The IC values of EO, ACE and HexE were 15.11\u202f±\u202f0.99, 118.16\u202f±\u202f1.7 and 324.26\u202f±\u202f2.45\u202fμg/mL, respectively, compared to IC of Tannic (23.83\u202f±\u202f1.9\u202fμg/mL) and butylated hydroxytoluene (BHT, 2.9\u202f±\u202f0.1\u202fμg/mL). Data suggested that the ripened fruits of S. terebinthifolius have potent antioxidant and antibacterial activities.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Constituents of leaves and flowers essential oils of Helichrysum pallasii (Spreng.) Ledeb. growing wild in Lebanon.

The chemical compositions of the essential oils obtained from leaves and flowers of Helichrysum pallasii were analyzed by gas chromatography and gas chromatography-mass spectrometry. Among the 102 identified constituents, hexadecanoic (16.2%), (Z,Z)-9,12-octadecadienoic (6.8%), tetradecanoic (2.6%), and (Z)-caryophyllene (4.2%) were the main constituent of the oil from leaves, while in the oil from flowers hexadecanoic (14.7%), (Z,Z)-9,12-octadecadienoic (14.2%), (Z)-caryophyllene (3.6%), and delta-cadinene (3.1%) predominated. The oils were both characterized by sesquiterpenes (33.4% for leaves and 33.7% for flowers, respectively) and and esters (30.3% in leaves and 35% in flowers, respectively). The in vitro activity of the essential oils of the plant against some microorganisms in comparison with chloramphenicol by the broth dilution method was determined. The oils exhibited a weak activity as inhibitors of growth of Staphylococcus epidermidis in vitro (minimum inhibitory concentration = 100 microg/mL).

Keyword: SCFA

Selective N-acylethanolamine-hydrolyzing amidase inhibition reveals a key role for endogenous palmitoylethanolamide in inflammation.

Identifying points of control in inflammation is essential to discovering safe and effective antiinflammatory medicines. Palmitoylethanolamide (PEA) is a naturally occurring lipid amide that, when administered as a drug, inhibits inflammatory responses by engaging peroxisome proliferator-activated receptor-alpha (PPAR-alpha). PEA is preferentially hydrolyzed by the cysteine amidase N-acylethanolamine-hydrolyzing amidase (NAAA), which is highly expressed in macrophages. Here we report the discovery of a potent and selective NAAA inhibitor, N-[(3S)-2-oxo-3-oxetanyl]-3-phenylpropanamide [(S)-OOPP], and show that this inhibitor increases PEA levels in activated leukocytes and blunts responses induced by inflammatory stimuli both in vitro and in vivo. These effects are stereoselective, mimicked by exogenous PEA, and abolished by PPAR-alpha deletion. (S)-OOPP also attenuates inflammation and tissue damage and improves recovery of motor function in mice subjected to spinal cord trauma. The results suggest that PEA activation of PPAR-alpha in leukocytes serves as an early stop signal that contrasts the progress of inflammation. The PEA-hydrolyzing amidase NAAA may provide a previously undescribed target for antiinflammatory medicines.

Keyword: SCFA

, essential oil, and phenolics modifications of black cumin fruit under NaCl stress conditions.

This research evaluated the effect of saline conditions on fruit yield, , and essential oils compositions and phenolics content of black cumin (Nigella sativa). This plant is one of the most commonly found aromatics in the Mediterranean kitchen. Increasing NaCl levels to 60 mM decreased significantly the fruits yield by 58% and the total amount by 35%. composition analysis indicated that linoleic was the major (58.09%) followed by oleic (19.21%) and (14.77%) . Salinity enhanced the linoleic percentage but did not affect the unsaturation degree of the pool and thus the oil quality. The essential oil yield was 0.39% based on the dry weight and increased to 0.53, 0.56, and 0.72% at 20, 40, and 60 mM NaCl. Salinity results on the modification of the essential oil chemotype from p-cymene in controls to γ-terpinene/p-cymene in salt-stressed plants. The amounts of total phenolics were lower in the treated plants. Salinity decreased mainly the amount of the major class, benzoics , by 24, 29, and 44% at 20, 40, and 60 mM NaCl. The results suggest that salt treatment may regulate bioactive compounds production in black cumin fruits, influencing their nutritional and industrial values.

Keyword: SCFA

Association between sn-2 fatty profiles of breast milk and development of the infant intestinal microbiome.

Increasing evidence shows that host diet and gut microbes are related. Previous studies have shown the effects of specific dietary fatty acids (FAs) on intestinal , but little is known about the effect of the stereospecifically numbered sn-2 position in triglycerides (TG) of human milk on the gut microbiome of infants. This study aimed at examining possible effects of sn-2 FAs of human milk on the gut microbial development of breastfeeding babies. Sn-2 FAs and intestinal were assessed by GC-MS and high-throughput 16S rRNA sequencing, respectively. The results showed that breast milk from mothers in China contained ten major sn-2 FAs dominated by (C, 54.42%), oleic (C n-9, 14.95%), linoleic (LA, C n-6, 12.81%), myristic (C, 4.50%) and C (3.17%). Total long chain unsaturated fatty acids (LCUFA) decreased from colostrum to mature milk, while total saturated fatty acids (SFA) showed no significant difference during lactation. A significant association between sn-2 FAs in milk and infant gut was found between decanoic (C), myristic (C), stearic (C), C, arachidonic (AA, C n-6), docosahexaenoic (DHA, C n-3) with Bacteroides, Enterobacteriaceae, Veillonella, Streptococcus, and Clostridium. These microbes were involved in short-chain fatty (SCFA) production and other functions, and significantly increased at 13-15 d after breastfeeding was initiated. C and DHA were relevant to most of the microbes. This study demonstrated the relatively steady profiles of sn-2 FAs in breast milk and gut of infants, together with their correlation during the breastfeeding period. The above results provided important information for designing the configuration of FAs in next-generation formulas for Chinese infants.

Keyword: SCFA

Process optimization and characterization of fragrant oil from red pepper (Capsicum annuum L.) seed extracted by subcritical butane extraction.

Red pepper seeds account for 450-500\u2009g\u2009kg of the total pepper weight and are often discarded as waste. In this study, process optimization and characterization of fragrant oil from roasted red pepper seed extracted by subcritical butane extraction were carried out.The optimal conditions of extraction were a temperature of 74.61\u2009°C, a time of 68.65\u2009min and a liquid/solid ratio of 30.24:1. The oil had a refractive index (25\u2009°C) of 1.471, a relative density of 0.900, an value of 1.421\u2009mg\u2009g oil, an iodine value of 127.035\u2009g per 100\u2009g, a saponification value of 184.060\u2009mg KOH g , an unsaponifiable matter content of 12.400\u2009g\u2009kg , a peroxide value of 2.465\u2009meq. O kg and a viscosity of 52.094 cP. The main in the oil were linoleic (72.95%) followed by (11.43%) and oleic (10.00%). The oil showed desirable thermal and oxidative stability. A total of 19 compounds, mostly aldehydes and alkenes, were identified from the oil.The results indicated that the method is appropriate for the preparation of fragrant red pepper seed oil, and the oil is suitable for used as edible oil. © 2016 Society of Chemical Industry.© 2016 Society of Chemical Industry.

Keyword: SCFA

Effect of Roasting Temperatures on the Properties of Bitter Apricot (Armeniaca sibirica L.) Kernel Oil.

compounds and quality changes of bitter apricot (Armeniaca sibirica L.) kernel oil (AKO) with different roasting conditions were determined. Bitter apricot kernels were roasted at 120, 130, 140 and 150°C for 15 min. Unroasted bitter apricot kernel oil was used as the control. Quality indicators included color, value and peroxide value, , total phenols and oxidative stability. Peroxide values of the tested oils were 0.46-0.82 meq/kg, values were 0.60-1.40 mg KOH/g, and total phenol contents were 54.1-71.5 μg GAE/g. Oleic was the major , followed by linoleic, , stearic and palmitoleic . Roasting increased the oxidative stability of bitter AKO. compounds were tentatively identified and semi-quantified. Among the 53 volatiles identified, benzaldehyde and benzyl alcohol were the major components. These two aroma compounds increased significantly during roasting and contributed sweet and almond flavors. Pyrazines were also prevalent and significantly increased with roasting. Sensory evaluation showed that roasted, nutty, sweet and oily aromas increased as roasting temperature increased.Practical applications: Bitter apricot kernels cannot be consumed directly, thus it is potentially beneficial to find uses for them, especially in China where bitter apricot processing is a significant industry. Roasted bitter AKO with a pleasant aroma could be prepared and might find use as an edible oil. The roasting process gave the bitter AKO a pleasant flavor. This study provided preliminary information on production parameters and potential quality control parameters.

Keyword: SCFA

Analysis of compounds and triglycerides of seed oils extracted from different poppy varieties (Papaver somniferum L.).

Poppy seed oil (Oleum Papaveris Seminis) is used for culinary and pharmaceutical purposes, as well as for making soaps, paints, and varnishes. Astonishingly, hardly anything was yet known about the compounds of this promising comestible. Likewise, there are no current published data about the triglyceride (TAG) composition of poppy seed oils available. In this investigation solid-phase microextraction (SPME) with DVB/Carboxen/PDMS Stable-Flex fiber was applied to the study of compounds of several seed oil samples from Papaver somniferum L. (Papaveraceae). 1-Pentanol (3.3-4.9%), 1-hexanal (10.9-30.9%), 1-hexanol (5.3-33.7%), 2-pentylfuran (7.2-10.0%), and caproic (2.9-11.5%) could be identified as the main compounds in all examined poppy seed oil samples. Furthermore, the TAG composition of these oils was analyzed by MALDI-ReTOF- and ESI-IT-MS/MS. The predominant TAG components were found to be composed of linoleic, oleic, and , comprising approximately 70% of the oils. TAG patterns of the different poppy varieties were found to be very homogeneous, showing also no significant differences in terms of the applied pressing method of the plant seeds.

Keyword: SCFA

Demonstration of bioprocess factors optimization for enhanced mono-rhamnolipid production by a marine Pseudomonas guguanensis.

We identified that Pseudomonas guguanensis produced macromolecular mono-rhamnolipid (1264.52\u202fDa) upon sensing n-hexadecane/diesel/kerosene from its surroundings. Permutation experiments were done to improve the laboratory-scale mono-rhamnolipid production (ie, a three-fold increase) using RSM validation. Consequently, maximal mono-rhamnolipids production [40-50\u202fmg/L] and emulsification abilities [65-70%] were encountered on day 8 using vegetable oil, peptone\u202f+\u202fyeast extract. EI values for the rhamnolipids were found to be 78±1.75% at 12.5\u202fmg/\u202fmL. Production and secretion of rhamnolipids were accompanied by aggregation of cells at day 6 as pictured in SEM. Pure monorhamnolipids of P. guguanensis was found to lower the surface tension of water to 32.98±0.3\u202fmN/m than the crude and CFSs of P. aeruginosa indicating efficient activity. Utilization and subsequent removal of hexadecane was 77.2% and the breakdown products were fatty acids [decanoic, hexadecanoic, octadecanoic acids and methyl stearates] as signified in Head-space GC-MS. The breakdown products of hexadecane are also present in the synthesized rhamnolipids suggesting their biosynthetic role. Rapid degradation of hexadecane, diesel and kerosene by this emulsifier combined with non-pathogenic trait of P. guguanensis identifies this organism as a viable option to remove n-alkanes from aquatic environments.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: SCFA

Microbial Quality of and Biochemical Changes in Fresh Soft, -Curd Xinotyri Cheese Made from Raw or Pasteurized Goat\'s Milk.

The microbiological quality of and changes in the main physicochemical parameters, together with the evolution of proteolysis, lipolysis and profiles of soft Xinotyri, a traditional Greek -curd cheese (pH≈4.4, moisture 65%, salt 1%) made from raw (RMC) or pasteurized (PMC) goat\'s milk without starters, were evaluated during aerobic storage at 4 C for 60 days. No statistically significant differences between the total nitrogen (TN) and nitrogen fraction (% of TN) contents, the degradation of intact α- or β-caseins, total free amino (FAA) contents, and the ratio of hydrophilic and hydrophobic peptides in the water-soluble fraction of RMC and PMC were found. Threonine, alanine and lysine were the principal FAAs. Oleic, , capric and caprylic , and ethyl hexonate, ethyl octanoate, ethyl decanoate, ethanol, 3-methyl butanol, phenyl ethyl alcohol and acetone were the most abundant free and compounds, respectively. Cheese lipolysis evolved slowly at 4 C, and milk pasteurization had no significant effect on it. Mesophilic lactic bacteria (LAB) were predominant in fresh cheese samples. PMC samples had significantly lower levels of enterococci and enterobacteria than RMC samples, while yeasts grew at similar levels during storage at 4 C. All cheese samples (25 g) were free of and . Coagulase-\u2028-positive staphylococci exceeded the 5-log safety threshold in fresh RMC samples, whereas they were suppressed (<100 CFU/g) in all PMC samples. Consequently, pasteurization of raw goat milk\'s and utilization of commercially defined or natural mesophilic LAB starters are recommended for standardizing the biochemical, microbial and safety qualities of fresh soft Xinotyri cheese.

Keyword: SCFA

Essential oils and composition of Tunisian and Indian cumin (Cuminum cyminum L.) seeds: a comparative study.

Cumin (Cuminum cyminum L.) seeds of two geographic origins, Tunisia (TCS) and India (ICS), were studied regarding their and essential oil composition.Oil yields were 17.77 and 15.40% for TCS and ICS respectively. Petroselinic (C18:1n-12) was the major in both varieties, with a higher proportion being found in TCS (55.90% of total (TFA)) than in ICS (41.42% TFA). Moreover, the most predominant were , petroselenic and linoleic , accounting for more than 91% TFA in both varieties. The unsaturated content was high: 70.95% TFA in TCS and 62.17% TFA in ICS. Essential oil yields differed significantly (P < 0.05) between the two varieties: 1.21 and 1.62% for ICS and TCS respectively. A total of 40 compounds were identified, 34 of which were present in both essential oils. The two varieties displayed different chemotypes: γ-terpinene/1-phenyl-1,2-ethanediol for TCS and cuminaldheyde/γ-terpinene for ICS.The study revealed that the biochemical composition of cumin seeds is origin-dependent and that cumin seeds are rich in an unusual , petroselinic . Besides, cumin essential oil is a rich source of many compounds, including cuminaldehyde and γ-terpinene. The overall results suggest the exploitation of cumin seeds as a low-cost renewable source for industrial processing in the fields of cosmetics, perfumes and pharmaceuticals.Copyright © 2011 Society of Chemical Industry.

Keyword: SCFA

High Amounts of n-Alkanes in the Composition of Asphodelus aestivus Brot. Flower Essential Oil from Cyprus.

There is only a couple of reports indicating essential oil composition of Asphodelus species in the literature. However, from the members of this genus many non- secondary metabolites were isolated. In Cyprus, Asphodelus aestivus Brot. can be found abundantly in all regions of the island. This plant has various ethnobotanical uses in Cyprus. There is no report on the volatiles nor the essential oil composition of A. aestivus. The smell of A. aestivus flowers resembles that of a cat pee which caught our attention. Therefore, we have carried out GC, GC/MS analysis of the essential oil (yield: 0.01 v/w) obtained from Asphodelus aestivus flowers. Seventeen compounds were identified in the essential oil comprising 96.2% of the oil. The major components of the essential oil were hexadecanoic 35.6%, pentacosane 17.4%, tricosane 13.4% and heptacosane 8.4%. In our results, we expected to see sulfur containing cat pee odorants due to the odor of the flower whereas high amounts of n-alkanes, saturated and minor amounts of acyclic diterpenes were observed.

Keyword: SCFA

components from flower-heads of Centaurea nicaeensis All., C. parlatoris Helder and C. solstitialis L. ssp. schouwii (DC.) Dostál growing wild in southern Italy and their biological activity.

The constituents of the flowerheads of Centaurea nicaeensis All., C. parlatoris Helder and C. solstitialis L. ssp. schouwii (DC.) Dostál were extracted by hydrodistillation and analysed by GC and GC-MS. Altogether 113 components were identified. and hydrocarbons were the most abundant components in the oils. Caryophyllene and caryophyllene oxide were the main compounds of the sesquiterpene fraction. The study on the biological activity of the oils shows no significant activity.

Keyword: SCFA

Fate of LCFA in the co-digestion of cow manure, food waste and discontinuous addition of oil.

Different concentrations of oily waste were added in a discontinuous mode and recurrently to anaerobic continuous stirred tank reactors fed with cow manure and food waste. Four continuous stirred tank reactors were run in parallel. A control reactor (R1) received no additional oil and R2, R3 and R4 received increasing concentrations of oil in two different experimental approaches. First, the lipids composition was forced to change suddenly, in three moments, without changing the total chemical oxygen demand (COD) fed to the reactors. The only long chain (LCFA) detected onto the R1 solid matrix was (C16:0). Nevertheless in the solid matrix of R2, R3 and R4C16:0 and stearic were detected. For occasional increase in the oil concentration up to 7.7gCOD(oil)/L(reactor) (55% Oil(COD)/Total(COD)) no statistical differences were detected between the reactors, in terms of methane production, effluent soluble COD, effluent and total and solids removal. Therefore this experiment allowed to conclude that cow manure-food waste co-digestion presents sufficient buffer capacity to endure solid-associated LCFA concentration up to 20-25gCOD-LCFA/kgTS. In a second experiment higher concentrations of oil were added, raising occasionally the concentration in the reactors to 9, 12, 15 and 18gCOD(oil)/L(reactor). All pulses had a positive effect in methane production, with the exception of the highest oil pulse concentration, that persistently impaired the reactor performance. This experiment demonstrates that threshold values for LCFA and C16:0 accumulation onto the solid matrix, of about 180-220gCOD-LCFA/kgTS and 120-150gCOD-C16:0/kgTS, should not be surpassed in order to prevent persistent reactor failure, as occurs in some full scale co-digestion plants.

Keyword: SCFA

Simultaneous Determination of Essential Oil Components and in Fennel using Gas Chromatography with a Polar Capillary Column.

Cultivated and wild growing samples of fennel (Foeniculum vulgare Mill., Apiaceae) from R. Macedonia were studied for their volatiles and composition. The main essential oil components isolated via hydrodistillation were: trans-anethole (>80%), estragole (< 6%), limonene (< 6%), anisaldehyde (< 1%) and 0.5 % fenchone. An alternative method for characterization of both the non-polar and non fractions was developed using n-hexane and dichloromethane (3:1, v/v) in a Soxhlet extraction followed by transesterification. The obtained extracts were then characterized and the dominant was 18:1 (petroselinic and oleic ) 75.0-82.8%, followed by 18:2 (linoleic ) 10.8-16.2% and other : (4.3-6.9%), stearic (1.2-1.7%) and myristic (0-2.9%). The results for the fraction after Soxhlet extraction and transesterification did not significantly differ from results obtained after hydrodistillation, especially for the main components (trans-anethole, estragole, fenchone and limonene), implying that the developed method can be used for simultaneous determination of volatiles and .

Keyword: SCFA

Suppression of adipogenesis by valproic through repression of USF1-activated synthesis in adipocytes.

VPA (valproic ), a short-chain that is a HDAC (histone deacetylase) inhibitor, is known to suppress adipogenesis. In the present study, we identified the molecular mechanism of VPA-mediated suppression of adipogenesis in adipocytes. VPA suppressed the accumulation of intracellular triacylglycerol. The expression levels of PPARγ (peroxisome-proliferator-activated receptor γ) and C/EBPα (CCAAT/enhancer-binding protein α), which are key regulators of adipogenesis, as well as the expression of SCD (stearoyl-CoA desaturase), were decreased by the treatment with VPA. Moreover, glycerol release was decreased in the VPA-treated cells, even though the transcription levels of ATGL (adipose triacylglycerol lipase), HSL (hormone-sensitive lipase) and MGL (monoacylglycerol lipase), all of which are involved in lipolysis, were elevated by the treatment with VPA. It is noteworthy that the expression level of FAS ( synthase) was significantly suppressed when the cells were cultured in medium containing VPA. Furthermore, VPA-mediated suppression of the accumulation of the intracellular triacylglycerols was prevented by the treatment with , a major product of FAS. The results of promoter-luciferase and chromatin immunoprecipitation assays demonstrated that USF1(upstream stimulating factor 1) bound to the E-box of the promoter region of the FAS gene. In addition, the expression of USF1 was decreased by the treatment with VPA. siRNA-mediated knockdown of the expression of the USF1 gene repressed adipogenesis along with the decreased expression of the FAS gene. The overexpression of USF1 enhanced both adipogenesis and the expression of FAS in VPA-treated cells. These results indicate that VPA suppressed adipogenesis through the down-regulation of USF1-activated synthesis in adipocytes.

Keyword: SCFA

Transformation of raw ewes\' milk applying "Grana" type pressed cheese technology: Development of extra-hard "Gran Ovino" cheese.

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes\' milk; and to produce a new typology of raw ewes\' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic bacteria (LAB) were in the range 10-10\u202fCFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 10\u202fCFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3\u202fh from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 10\u202fCFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese were , oleic and myristic and the saturated /unsaturated ratio was 2.17. Forty-one compounds, including , esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: SCFA

[Study on fat-soluble components in different parts of miao ethnomedicine Pileostegia viburnoides by the herbal blitzkrieg extractor].

To study the fat-soluble components in different parts of Pileostegia viburnoides.The fat-soluble components in different parts of Pileostegia viburnoides were extracted by Herbal Blitzkrieg Extractor (HBE), and its constituents were analyzed by GC-MS.37 compounds from the stems, 14 compounds from the leaves were separated and identified. The ratios of identification of total compounds were 90. 63% and 81.61%, respectively.This result provides the scientific basis for ex-ploiting resources of Miao ethnomedicine Pileostegia viburnoides.

Keyword: SCFA

[Pyrolysis of the Lysimachia foenum-graecum Hance extract by online pyrolysis-gas chromatography-mass spectrometry].

In order to study the pyrolytic properties of Lysimachia foenum-graecum Hance extract, it was pyrolysed and detected by online pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). The pyrolytic experimental conditions were designed to simulate the real combustion conditions inside a burning cigarette. The sample was heated at 30 degrees C/s from 300 to 900 degrees C (held for 5 s) under the flow of 9% oxygen in nitrogen. The pyrolytic components and components were compared. The results showed that 64 pyrolytic components were detected, with 88.27% of the total peak area, including linoleic ethyl ester (10.33%), hexadecanoic , ethyl ester (9.12%), 9,12,15-octadecatrienoic , (Z,Z,Z) - (8.03%), 2-furan-carboxaldehyde, 5-(hydroxymethyl) - (6.02%), neophytadiene (5.12%), heptadecanoic , ethyl ester (4.50%), acetic , phenyl ester (3.51%), 5-methoxy-2, 2-di-methylindan-1-one (2.73%). The number of pyrolytic components was more than that of the components, and 20 components were identified in both pyrolytic components and components, including higher and their esters, neophytadiene, 2-furancarboxaldehyde, (hydroxymethyl)-, and 2 (5H)-furanone, 3-hydroxy-4, 5-dimethyl-. The on-line pyrolysis was similar to the real cigarette combustion conditions. The method is a simple, rapid and good qualitative method for the pyrolysis.

Keyword: SCFA

Physiological and molecular genetic studies on two elicitors for improving the tolerance of six Egyptian soybean cultivars to cotton leaf worm.

Cotton leaf worm (Spodoptera littoralis) is considered one of the most destructive agricultural pests in Egypt. Six soybean cultivars (Giza-21, Giza-22, Giza-35, Giza-82, Giza-83 and Giza-111) were grown under natural infection with cotton leaf worm. The effect of two elicitors, methyl jasmonate and sodium nitroprusside on enhancing the ability of susceptible cultivars to tolerate (Spodoptera littoralis) was studied. Giza-35 and Giza-111 showed tolerance performance under natural infection compared to Giza-22 and Giza-82 as sensitive ones, while Giza-83 and Giza-21 showed moderate tolerance. Both treatments positively affected seed yield and its components and composition. Extracted showed variable changes in treated plants compared with the untreated controls. Plants treated with the two elicitors showed an increase in Linoleic and Linolenic and decrease in and Palmitolic content. Treatment with methyl jasmonate was found to be more effective than sodium nitroprusside and enhanced resistance of the susceptible cultivars. Eight IRAP and iPBS retrotransposon-based markers were used to detect genetic differences among studied soybean cultivars and to develop molecular genetic markers for cotton leaf worm infestation. The technique successfully identified soybean genotypes in addition to nineteen molecular markers related to soybean tolerance.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: SCFA

Emex spinosa (L.) Campd. ethyl acetate fractions effects on inflammation and oxidative stress markers in carrageenan induced paw oedema in mice.

Emex spinosa (L.) Campd. (E. spinosa) locally known as "hillaioua" has always been used in folk medicine for the treatment of inflammation and pain. It is still being exploited by pharmaceutical companies for its potential remedial effects.In this study, the effects of E. spinosa (L.) Campd. against acute inflammation, pain and oxidative damage were evaluated.Total phenols and flavonoids were evaluated. Anti-inflammatory and analgesic activities the E. spinosa ethyl acetate fractions of the aerial (Es EtOAc-AP) and underground (Es EtOAc-R) parts were assessed on carrageenan-induced paw oedema (100\u202fmg/kg BW) and acetic -induced writhing response (50, 100 and 150\u202fmg/kg BW), respectively. The E. spinosa fractions effects on oxidative stress markers and inflammatory parameters were determined. Gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify various chemical components.The ethyl acetate fractions were shown to be the most active thanks to their phenolic and flavonoid contents richness. Intraperitoneal administration of E. spinosa ethyl acetate fractions at 100\u202fmg/kg BW, one hour before carrageenan injection, significantly inhibited the oedema formation by 89.31% and 97.7% for the aerial and underground parts respectively when compared to the reference drug "dexamethasone"\u202f(51.9%). Besides, a significant increase (p\u202f≤\u202f0.001) of the dermal antioxidant enzymes (the superoxide dismutase (SOD)), catalase (CAT) and glutathione peroxidase (GPx) was observed five hours after carrageenan administration. The best restoration was obtained with Es EtOAc-R (82.04%, 93.55% and 93.55% respectively for SOD, CAT and GPx activities). Moreover, EtOAc-fractions treated mice proved their ability to restore both of CRP and fibrinogen (p\u202f<\u202f0.001). In addition, E. spinosa EtOAc-fractions attenuated abdominal contractions (p\u202f<\u202f0.05) by 71.69% and 82.41% for the aerial part and roots respectively at 150\u202fmg/kg BW against 100% for dichlofenac sodium used as standard drug. The phytochemical analysis of Es EtOAc-AP and Es EtOAc-R by GC-MS may explain the obtained results. The analysis of the fractions demonstrated the presence of and linoleic known for their anti-inflammatory and analgesic capacities.These findings explain the traditional use of E. spinosa in folk medicine and suggest that E. spinosa fractions could be a promising herbal drug.Copyright © 2018. Published by Elsevier B.V.

Keyword: SCFA

Compared with stearic , increased the yield of milk fat and improved feed efficiency across production level of cows.

The effects of dietary and stearic on feed intake, yields of milk and milk components, and feed efficiency of dairy cows were evaluated in an experiment with a crossover arrangement of treatments with a covariate period. Cows with a wide range of milk production (38 to 65 kg/d) were used to determine if response to fat supplementation varied according to production level. Thirty-two Holstein cows (143 ± 61 d in milk) were assigned randomly to a treatment sequence within level of milk production. Treatments were diets supplemented (2% of diet dry matter) with (PA; 97.9% C16:0) or stearic (SA; 97.4% C18:0). Treatment periods were 21 d and cows were fed a nonfat supplemented diet for 14 d immediately before the first treatment period. The final 4d of each period were used for sample and data collection. Milk production measured during the covariate period (preliminary milk yield) was used as the covariate. No interactions were detected between treatment and preliminary milk yield for the production response variables measured. Compared with SA, the PA treatment increased milk fat concentration (3.66 vs. 3.55%) and yield (1.68 vs. 1.59 kg/d), and 3.5% fat-corrected milk yield (47.5 vs. 45.6 kg/d). Treatment did not affect dry matter intake, milk yield, milk protein yield, body weight, or body condition score. Milk protein concentration was lower for PA compared with SA treatment (3.24 vs. 3.29%). The PA treatment increased feed efficiency (3.5% fat-corrected milk yield/dry matter intake) compared with SA (1.48 vs. 1.40). The increase in milk fat yield by PA was entirely accounted for by a 24% increase in 16-carbon output into milk. Yields of de novo (3.2%) and preformed (2.9%) were only slightly decreased by PA relative to SA. The PA treatment increased plasma concentration of nonesterified (96.3 vs. 88.2 μEq/L) and glucose (56.6 vs. 55.7 mg/dL) compared with SA, but insulin and β-hydroxybutyrate were not altered by the treatments. Results demonstrate that is more effective than stearic in improving milk fat concentration and yield as well as efficiency of feed conversion to milk. Responses were independent of production level and without changes in body condition score or body weight. Further studies are required to test the consistency of these responses across different types of diets.Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Aframomum stipulatum (Gagnep) K. Schum and Aframomum giganteum (Oliv. & Hanb) K. Schum as Aroma Tincto Oleo Crops resources: essential oil, , sterols, tocopherols, and tocotrienols composition of different fruit parts of Congo varieties.

Today, few known plant species provide both an essential oil (EO) and a vegetable oil (VO). Seed and husk of two Aframomum species were investigated and compared in terms of EO, , tocopherols, and tocotrienols.EO yield reaches 15.3 g kg(-1) in the seeds and 3.2 g kg(-1) in the husks, while VO yield is 180.0 g kg(-1) in the seeds and 25.0 g kg(-1) in the husks. β-Pinene, 1,8-cineol, α-selinene, terpine-4-ol, linalool, myrtenal and β-caryophyllene are the major compounds of seed and husk EO. analysis of two Aframomum species shows that oleic, linoleic, and were the major compounds of VO. Total sterol contents reached 4.3 g kg(-1) in seed VO and 8.5 g kg(-1) in husk VO. An appreciable amount of tocopherols (0.52 g kg(-1) ) was found in seed VO.The seed and husk oil of A. stipulatum and A. giganteum fruits are rich sources of many bioactive constituents such as , sterols, tocopherols and tocotrienols. These tropical wild fruits can be considered as new Aroma Tincto Oleo Crops (ATOC) resources that contain both EOs and VOs.Copyright © 2012 Society of Chemical Industry.

Keyword: SCFA

Does valproate therapy in epileptic patients contribute to changing atherosclerosis risk factors? The role of lipids and free .

We aimed to demonstrate the relationship between the valproate (VPA) treatment versus lipid and serum free (FFAs) profiles to be the potential atherosclerosis risk factor in epileptic patients.Fasting blood samples were taken from 21 adult VPA-treated patients and 21 controls. The profiles of lipids, FFAs, clinical parameters and body mass index (BMI) were evaluated.No significant differences between the study group and controls were found for any of the studied parameters. However, significant differences in the total cholesterol (CHOL), low-density-lipoprotein cholesterol (LDL), triglycerides, the CHOL/HDL (high-density-lipoprotein cholesterol) ratio, and Atherogenic Index of Plasma were observed for overweight patients when compared to those of normal weight. Patients with uncontrolled epilepsy tended to have significantly lower level than seizure-free patients. Oleic was found to be positively correlated with VPA concentration for patients with uncontrolled epilepsy, and with the dose corrected VPA concentration for all the patients. The was however negatively correlated with stearic for both the controls and the patients with uncontrolled epilepsy. PLS method revealed CHOL, LDL, triglycerides and myristic to be positively interrelated for the whole group under the study, whereas these parameters were found to be negatively correlated with VPA concentration, and positively with BMI. Furthermore, high sensitivity C-reactive protein was found to be negatively correlated with levels.Overweight VPA-treated patients are exposed to higher risk of atherosclerosis. Alterations in FFAs are likely to depend on seizures control, and on VPA levels.Copyright © 2016. Published by Elsevier Urban & Partner Sp. z o.o.

Keyword: SCFA

1\'-Acetoxychavicol acetate ameliorates age-related spatial memory deterioration by increasing serum ketone body production as a complementary energy source for neuronal cells.

1\'-Acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Alpinia galangal. Here, we examined the effect of ACA on learning and memory in senescence-accelerated mice prone 8 (SAMP8). In mice that were fed a control diet containing 0.02% ACA for 25 weeks, the learning ability in the Morris water maze test was significantly enhanced in comparison with mice that were fed the control diet\xa0alone. In the Y-maze test, SAMP8 mice showed decreased spontaneous alterations in comparison with senescence-accelerated resistant/1 (SAMR1) mice, a homologous control, which was improved by ACA pretreatment. Serum metabolite profiles were obtained by GC-MS analysis, and each metabolic profile was plotted on a 3D score plot. Based upon the diagram, it can be seen that the distribution areas for the three groups were completely separate. Furthermore, the contents of β-hydroxybutyric and in the serum of SAMP8-ACA mice were higher than those of SAMP8-control mice and SAMR1-control mice. We also found that SAMR1 mice did not show histological abnormalities, whereas histological damage in the CA1 region of the hippocampus in SAMP8-control mice was observed. However, SAMP8-ACA mice were observed in a similar manner as SAMR1 mice. These findings confirm that ACA increases the serum concentrations of β-hydroxybutyric and levels and thus these fuels might contribute to the maintenance of the cognitive performance of SAMP8 mice.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: SCFA

Soraphen, an inhibitor of the acetyl-CoA carboxylase system, improves peripheral insulin sensitivity in mice fed a high-fat diet.

Inhibition of the acetyl-CoA carboxylase (ACC) system, consisting of the isozymes ACC1 and ACC2, may be beneficial for treatment of insulin resistance and/or obesity by interfering with de novo lipogenesis and beta-oxidation. We have evaluated effects of pharmacological inhibition of ACC by soraphen (SP) on high fat (HF) diet-induced insulin resistance in mice.Male C57Bl6/J mice were fed control chow, a HF diet or a HF diet supplemented with SP (50 or 100 mg/kg/day).Body weight gain and total body fat content of SP-treated animals were significantly reduced compared with HF-fed mice. Fractional synthesis of palmitate was significantly reduced in mice treated with SP, indicative for ACC1 inhibition. Plasma beta-hydroxybutyrate levels were significantly elevated by SP, reflecting simultaneous inhibition of ACC2 activity. Mice treated with SP showed improved peripheral insulin sensitivity, as assessed by hyperinsulinaemic euglycaemic clamps.Pharmacological inhibition of the ACC system is of potential use for treatment of key components of the metabolic syndrome.

Keyword: SCFA

Rapid detection of radiation-induced hydrocarbons in cooked ham.

Solid phase microextraction (SPME) coupled with either gas chromatography-ionization flame detector (CG-FID) or multidimensional gas chromatography-mass spectrometry (MDGC-MS) was evaluated for its ability to detect hydrocarbons produced during the irradiation of cooked ham. The chromatogram of an irradiated sample obtained using GC-FID showed a complex pattern of peaks, with several co-eluting peaks superimposed, indicating that the method was unlikely to resolve adequately the hydrocarbons formed during irradiation. Using SPME-MDGC-MS 1-tetradecene (C(1-14:1)), n-pentadecane (C(15:0)), 1-hexadecene (C(1-16:1)), n-heptadecane (C(17:0)) and 8-heptadecene (C(8-17:1)) were detected in cooked ham irradiated at 0.5, 2, 4 and 8kGy. This method allows the detection of most n-alkanes and n-alkenes produced during the irradiation of the majority of in cooked ham, namely oleic , stearic and . SPME is rapid and inexpensive and does not require organic solvents. The proposed SPME-MDGC-MS method allows the determination of radiolytic markers in cooked ham in less than 115min.Copyright © 2011 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Acetate anion-triggered peroxygenation of non-native substrates by wild-type cytochrome P450s.

Cytochrome P450SPα (P450SPα) and cytochrome P450BSβ (P450BSβ) belonging to the CYP152 family of enzymes (CYP152s) can utilize H2O2 efficiently as an oxidant for the generation of compound I. Although P450SPα and P450BSβ have very high substrate specificity and catalyse hydroxylation of long-chain exclusively, we found that they can oxidize non-native substrates such as styrene simply by including medium chain length n-alkyl carboxylic as "decoy molecules." Although we had assumed that acetic did not serve as a decoy molecule, P450SPα and P450BSβ efficiently catalysed oxidation of non-native substrates when the reaction was carried out at a high concentration of acetate anion. The turnover rate for epoxidation of styrene catalysed by P450BSβ in the presence of 1 M acetate anion reached 590 ± 30 min(-1).

Keyword: SCFA

Agonist-induced activation releases peroxisome proliferator-activated receptor beta/delta from its inhibition by palmitate-induced nuclear factor-kappaB in skeletal muscle cells.

The mechanisms by which elevated levels of free cause insulin resistance are not well understood, but there is a strong correlation between insulin resistance and intramyocellular lipid accumulation in skeletal muscle. In addition, accumulating evidence suggests a link between inflammation and type 2 diabetes. The aim of this work was to study whether the exposure of skeletal muscle cells to palmitate affected peroxisome proliferator-activated receptor (PPAR) beta/delta activity. Here, we report that exposure of C2C12 skeletal muscle cells to 0.75 mM palmitate reduced (74%, P<0.01) the mRNA levels of the PPARbeta/delta-target gene pyruvatedehydrogenase kinase 4 (PDK-4), which is involved in utilization. This reduction was not observed in the presence of the PPARbeta/delta agonist L-165041. This drug prevented palmitate-induced nuclear factor (NF)-kappaB activation. Increased NF-kappaB activity after palmitate exposure was associated with enhanced protein-protein interaction between PPARbeta/delta and p65. Interestingly, treatment with the PPARbeta/delta agonist L-165041 completely abolished this interaction. These results indicate that palmitate may reduce utilization in skeletal muscle cells by reducing PPARbeta/delta signaling through increased NF-kappaB activity.

Keyword: SCFA

[Chemical constituents of ethyl acetate fraction of Tetrastigma planicaule].

To investigate the chemical constituents of ethyl acetate of Tetrastigma planicaule.The ethyl acetate fraction was isolated and purified by repeated silica gel chromatography and recrystallization. The structures of compounds were identified by physical-chemical methods and spectral data such as MS, 1H-NMR, 13C-NMR, etc.Six compounds were identified as gheddic (I), tricosanol (II), beta-sitosterol (III), (IV), ethyl gallate (V) and vanillin (VI), respectively.Compounds I, II, V and VI are isolated from this plant for the first time.

Keyword: SCFA

Valproic selectively inhibits conversion of arachidonic to arachidonoyl-CoA by brain microsomal long-chain acyl-CoA synthetases: relevance to bipolar disorder.

Several drugs used to treat bipolar disorder (lithium and carbamazepine), when administered chronically to rats, reduce the turnover of arachidonic , but not docosahexaenoic , in brain phospholipids by decreasing the activity of an arachidonic -selective phospholipase A(2). Although chronic valproic produces similar effects on brain arachidonic and docosahexaenoic turnover, it does not alter phospholipase A(2) activity, suggesting that it targets a different enzyme in the turnover pathway.By isolating rat brain microsomal long-chain acyl-CoA synthetases (Acsl), we show in vitro that valproic is a non-competitive inhibitor of Acsl, as it reduces the maximal velocity of the reaction without changing the affinity of the substrate for the enzyme. While valproic inhibited the synthesis of arachidonoyl-CoA, palmitoyl-CoA, and docosahexaenoyl-CoA, the K (i )for inhibition of arachidonoyl-CoA synthesis (14.1 mM) was approximately one fifth the K (i) for inhibiting palmitoyl-CoA (85.4 mM) and docosahexaenoyl-CoA (78.2 mM) synthesis. As chronic administration of valproic in bipolar disorder achieves whole-brain levels of 1.0 to 1.5 mM, inhibition of arachidonoyl-CoA formation can occur at brain concentrations that are therapeutically relevant to this disease. Furthermore, brain microsomal Acsl did not produce valproyl-CoA.This study shows that valproic acts as a non-competitive inhibitor of brain microsomal Acsl, and that inhibition is substrate-selective. The study supports the hypothesis that valproic acts in bipolar disorder by reducing the brain arachidonic cascade, by inhibiting arachidonoyl-CoA formation.

Keyword: SCFA

Effect of metabolic stressors on survival and growth of in\xa0vitro cultured ovine preantral follicles and enclosed oocytes.

The present study was undertaken to study the effect of metabolic stressors like elevated levels of ammonia, urea, Non-esterified (NEFA) and β-hydroxybutyric (BHB) on preantral follicle growth, survival, growth rates of oocytes enclosed in preantral follicles (PFs), maturation rates of oocytes recovered from cultured follicles, hormone production (estrogen and progesterone), reactive oxygen species (ROS) as well as superoxide dismutase (SOD) activity. Small pre-antral follicles (SPFs, 100-250\xa0μm) and large pre-antral follicles (LPFs, 250-450\xa0μm) were isolated from slaughterhouse ovaries by a mechanical cum enzymatic method. SPFs and LPFs were cultured in\xa0vitro for 14 and 7 days respectively and examined for their growth, survival and growth rates of enclosed oocytes in PFs exposed with different concentration of ammonia (0, 100, 150, 200, 250, 300 and 400\xa0μM), urea (0, 4, 4.5, 5, 5.5,6, 7 and 8\xa0mM), NEFA [Basal NEFA (70\xa0μM): stearic , SA (10\xa0μM)+, PA(20\xa0μM)+oleic , OA(40\xa0μM), b) Medium combo (140\xa0μM): SA (20\xa0μM)+ PA(40\xa0μM)+ OA(80\xa0μM), c) High combo (210\xa0μM): SA (30\xa0μM)+PA(60\xa0μM)+OA(120\xa0μM), d) Very high Combo (280\xa0μM): SA(40\xa0μM)+PA(80\xa0μM)+OA(160\xa0μM)] and BHB (0, 0.5, 0.75, and 1\xa0μM). Results indicated that ammonia, urea, NEFA and BHB caused inhibition of survival and growth of in\xa0vitro cultured ovine PFs and enclosed oocytes at the levels of 300\xa0μM, 8\xa0mM, high combo level of NEFA and 0.75\xa0μM respectively. Our study may contribute to the identification of the mechanisms involved in decline of fertility due to metabolic and nutritional stress in ruminants.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: SCFA

Physicochemistry, microbiology, composition and profile of traditional Söğle tulum (goat\'s skin bag) cheese.

Söğle cheese is a type of traditional tulum (goat\'s skin bag) cheese produced from goat\'s milk or a mixture of goat\'s and sheep\'s milk in Küçük Söğle and Büyük Söğle villages, Antalya, Turkey. This is the first study that aimed to investigate the physicochemistry, microbiology, composition and profile of traditional Söğle cheeses ripened in goat\'s skins for three months in wells. The pH and titratable acidity values, total solids, fat, protein and salt contents of the Söğle cheese samples ranged from 4.9 to 5.5, from 1.4 to 2.6%, from 46.5 to 55.0%, from 2.0 to 4.5%, from 34.9 and 42.2% and from 4.1 to 8.2%, respectively. The highest proportion of in all cheese samples was (C16:0). The lactobacillus, lactococcus, total aerobic mesophilic bacteria and yeast and mold counts in the cheese samples were found to be between 7.1 and 8.5 log cfu/g, 7.8 and 8.7 log cfu/g, 7.6 and 8.5 log cfu/g and 1.0 and 4.8 log cfu/g, respectively. Fifty-three components were identified in the cheese samples including 13 esters, 7 ketones, 5 , 2 aldehydes, 5 alcohols, 12 terpenes, and 9 miscellaneous compounds. Ketones and terpenes were the predominant compounds.

Keyword: SCFA

New aristolochic and other chemical constituents of Aristolochia maurorum growing wild in Jordan.

Investigation of the chemical constituents of Aristolochia maurorum growing wild in Jordan resulted in the isolation and characterisation of one new compound in addition to 19 known compounds. The new compound was identified as aristolochic II alanine amide (14). The other known compounds were the following: (1), β-sitosterol (2), E-ethyl-p-coumarate (3), Z-ethyl-p-coumarate (4), aristolochic IV methyl ester (5), aristolactam I (6), loliolide (7), (+)-dehydrovomifoliol (8), glycerol-1-palmitate (9), aristolochic I (10), E-p-coumaric (11), E-N-coumaroyltyramine (12), β-sitosteryl glucoside (13), aristolochic IV (15), aristolochic III (16), esculetin (17), uracil (18), shepherdine (19) and adenosine (20). The isolated compounds were characterised by different spectroscopic methods including NMR (1D and 2D), UV, IR and HRESIMS.

Keyword: SCFA

Characterization of Fatty , Amino and Volatile Compound Compositions and Bioactive Components of Seven Coffee (Coffea robusta) Cultivars Grown in Hainan Province, China.

Compositions of fatty , amino acids, and volatile compound were investigated in green coffee beans of seven cultivars of Coffea robusta grown in Hainan Province, China. The chlorogenic acids, trigonelline, caffeine, total lipid, and total protein contents as well as color parameters were measured. Chemometric techniques, principal component analysis (PCA), hierarchical cluster analysis (HCA), and analysis of one-way variance (ANOVA) were performed on the complete data set to reveal chemical differences among all cultivars and identify markers characteristic of a particular botanical origin of the coffee. The major fatty acids of coffee were linoleic , , oleic , and arachic . Leucine (0.84 g/100 g DW), lysine (0.63 g/100 g DW), and arginine (0.61 g/100 g DW) were the predominant essential amino acids (EAAs) in the coffee samples. Seventy-nine volatile compounds were identified and semi-quantified by HS-SPME/GC-MS. PCA of the complete data matrix demonstrated that there were significant differences among all cultivars, HCA supported the results of PCA and achieved a satisfactory classification performance.

Keyword: SCFA

Using [H]water to quantify the contribution of de novo palmitate synthesis in plasma: enabling back-to-back studies.

An increased contribution of de novo lipogenesis (DNL) may play a role in cases of dyslipidemia and adipose accretion; this suggests that inhibition of synthesis may affect clinical phenotypes. Since it is not clear whether modulation of one step in the lipogenic pathway is more important than another, the use of tracer methods can provide a deeper level of insight regarding the control of metabolic activity. Although [H]water is generally considered a reliable tracer for quantifying DNL in vivo (it yields a homogenous and quantifiable precursor labeling), the relatively long half-life of body water is thought to limit the ability of performing repeat studies in the same subjects; this can create a bottleneck in the development and evaluation of novel therapeutics for inhibiting DNL. Herein, we demonstrate the ability to perform back-to-back studies of DNL using [H]water. However, this work uncovered special circumstances that affect the data interpretation, i.e., it is possible to obtain seemingly negative values for DNL. Using a rodent model, we have identified a physiological mechanism that explains the data. We show that one can use [H]water to test inhibitors of DNL by performing back-to-back studies in higher species [i.e., treat nonhuman primates with platensimycin, an inhibitor of synthase]; studies also demonstrate the unsuitability of [C]acetate.

Keyword: SCFA

Characterisation of Fecal Soap Fatty Acids, Calcium Contents, Bacterial Community and Short-Chain Fatty Acids in Sprague Dawley Rats Fed with Different sn-2 Triacylglycerols Diets.

The structure of dietary triacylglycerols is thought to influence fatty and calcium absorption, as well as intestinal population of the host. In the present study, we investigated the impact of (PA) esterified at the sn-2 position on absorption of fatty and calcium and composition of intestinal microorganisms in rats fed high-fat diets containing either low sn-2 PA (12.1%), medium sn-2 PA (40.4%) or high sn-2 PA (56.3%), respectively. Fecal fatty profiles in the soaps were measured by gas chromatography (GC), while fecal calcium concentration was detected by ICP-MS. The fecal microbial composition was assessed using a 16S rRNA high-throughput sequencing technology and fecal short-chain fatty acids were detected by ion chromatograph. Dietary supplementation with a high sn-2 PA fat significantly reduced total fecal contents of fatty acids soap and calcium compared with the medium or low sn-2 PA fat groups. Diet supplementation with sn-2 PA fat did not change the entire profile of the gut community at phylum level and the difference at genera level also were minimal in the three treatment groups. However, high sn-2 PA fat diet could potentially improve total short-chain fatty acids content in the feces, suggesting that high dietary sn-2 PA fat might have a beneficial effect on host intestinal health.

Keyword: SCFA

Influence of calcium chloride in the high temperature acetification by strain Acetobacter aceti WK for vinegar.

To improve the thermotolerant properties (TTP) of acetic bacteria (AAB) cells for high temperature acetification.At high temperature (36 ± 1°C), the acetification rate (ETA) is usually lower than at 30 ± 1°C. The addition of 0·15% calcium chloride (CaCl2 ) may decrease the negative effect of the increase of temperature from 30 ± 1°C to 36 ± 1°C on the ETA. The effect of CaCl2 on the thermotolerant properties of acetic bacteria cells was investigated. The CaCl2 increased the content of phospholipids (phosphotidylcholine and phosphatidylglycerol), (cis-vaccenic , and myristic ) and the activities of membrane-bound enzymes involved in acetification, alcohol dehydrogenase and aldehyde dehydrogenase. Transmission electron microscope images revealed a more compact cell wall with CaCl2. Process consistency at 36 ± 1°C was tested in nine sequential acetification cycles using 0·15% (w/v) CaCl2. High ETAs (9·33 ± 0·6; 8·67 ± 0·8 and 9·67 ± 0·7 g l(-1) day(-1)) were obtained during the last three cycles.The results confirm that changes of the content of lipid, activities of membrane-bound enzymes and cell-wall thickness occurred with added CaCl2.High temperature acetification (HTA) with additions of CaCl2 was investigated. Significant reductions in the overall production costs result from lower cooling costs associated with HTA.© 2015 The Society for Applied Microbiology.

Keyword: SCFA

Selective cannabinoid-1 receptor blockade benefits and triglyceride metabolism significantly in weight-stable nonhuman primates.

The goal of this study was to determine whether administration of the CB₁ cannabinoid receptor antagonist rimonabant would alter flux in nonhuman primates. Five adult baboons (Papio Sp) aged 12.1 ± 4.7 yr (body weight: 31.9 ± 2.1 kg) underwent repeated metabolic tests to determine and TG flux before and after 7 wk of treatment with rimonabant (15 mg/day). Animals were fed ad libitum diets, and stable isotopes were administered via diet (d₃₁-tripalmitin) and intravenously (¹³C₄-palmitate, ¹³C₁-acetate). Plasma was collected in the fed and fasted states, and blood lipids were analyzed by GC-MS. DEXA was used to assess body composition and a hyperinsulinemic euglycemic clamp used to assess insulin-mediated glucose disposal. During the study, no changes were observed in food intake, body weight, plasma, and tissue endocannabinoid concentrations or the quantity of liver-TG originating from de novo lipogenesis (19 ± 6 vs. 16 ± 5%, for pre- and posttreatment, respectively, P = 0.39). However, waist circumference was significantly reduced 4% in the treated animals (P < 0.04), glucose disposal increased 30% (P = 0.03), and FFA turnover increased 37% (P = 0.02). The faster FFA flux was consistent with a 43% reduction in these used for TRL-TG synthesis (40 ± 3 vs. 23 ± 4%, P = 0.02) and a twofold increase in TRL-TG turnover (1.5 ± 0.9 vs. 3.1 ± 1.4 μmol·kg⁻¹·h⁻¹, P = 0.03). These data support the potential for a strong effect of CB₁ receptor antagonism at the level of adipose tissue, resulting in improvements in fasting turnover of at the whole body level, central adipose storage, and significant improvements in glucose homeostasis.

Keyword: SCFA

Isolation of alpha-glucosidase inhibitors produced by an endophytic fungus, Colletotrichum sp. TSC13 from Taxus sumatrana.

Colletotrichum sp. have potential to act as antidiabetic agent, due to its alpha-glucosidase inhibitory. Therefore, the objective of present study was to isolate and identify the bioactive compounds responsible for the alpha-glucosidase inhibitory activity in Colletotrichum sp. TSC13. The methanol extract of TSC13 mycelia, was partitioned with n-hexane, chloroform and ethyl acetate. The n-hexane fraction exhibited the strongest alpha-glucosidase inhibitory activity. Column chromatography of this fraction resulted in 8 sub-fractions (F1-8). Fraction 3 (F3) which showed 71.4 +/- 2.4% inhibition was analysed further. Analysis using GC-MS after methylation of F3 and comparison to spectra databases and confirmation using authentic sample standards showed that F3 had two saturated methyl esters, and stearic methyl esters and three unsaturated methyl esters, oleic , linoleic and linoleinic methyl esters. Unsaturated showed higher activity than the saturated and the methyl esters form of unsaturated showed slightly less active than the free . Further analysis using an ethyl acetate extract, it was confirmed that most of the were present in the form of free . Therefore, it was concluded that the alpha-glucosidase inhibitor compounds in Colletotrichum sp. TSC13 were unsaturated . This is the first report that a Colletotrichum sp. from T. sumatrana has alpha-glucosidase inhibitory activity.

Keyword: SCFA

Chemically transformable configurations of mercaptohexadecanoic self-assembled monolayers adsorbed on Au(111).

Carboxyl-terminated self-assembled monolayers (SAMs) are commonly used in a variety of applications, with the assumption that the molecules form well-ordered monolayers. In this work, near-edge X-ray absorption fine structure measurements verify that well-ordered monolayers can be formed using acetic in the solvent. Disordered monolayers with unbound molecules present in the film result using only ethanol. A stark reorientation occurs upon deprotonation of the end group by rinsing in a KOH solution. This reorientation of the end group is reversible with tilted-over, hydrogen-bound carboxyl groups while the carboxylate ion end groups are upright. C(1s) photoemission shows that SAMs formed and rinsed with acetic in ethanol have protonated end groups, while SAMs formed without acetic have a large fraction of carboxylate-terminated molecules.

Keyword: SCFA

Young women partition towards ketone body production rather than VLDL-TAG synthesis, compared with young men.

Before the menopause, women are relatively protected against CVD compared with men. The reasons for this sex difference are not completely understood, but hepatic metabolism may play a role. The present study aimed to investigate the utilisation of plasma NEFA by the liver and to determine whether they are partitioned differently into ketone bodies and VLDL-TAG in healthy, lean young men and women. Volunteers were studied during a prolonged overnight fast (12-19 h) using an intravenous infusion of [U-¹³C]palmitate. After 12 h fasting, the women had a more advantageous metabolic profile with lower plasma glucose (P < 0·05) and TAG (P < 0·05) but higher plasma NEFA (P < 0·05) concentrations. Plasma 3-hydroxybutyrate (3-OHB) concentrations rose more in women than in men, and the transfer of ¹³C from [U-¹³C]palmitate to plasma [¹³C]3-OHB reached a plateau 6-7 h after the start of the infusion in women but was still increasing at 6 h in men. This implies a slower 3-OHB production rate and/or dilution by other precursor pools in men. In women, the high isotopic enrichment of plasma 3-OHB suggested that systemic plasma were the major source of 3-OHB production. However, in men, this was not observed during the course of the study (P < 0·01). There were no sex differences for the incorporation of ¹³C into VLDL1- or VLDL2-TAG. The ability of young women to partition towards ketone body production rather than VLDL-TAG may contribute to their more advantageous metabolic profile compared with young men.

Keyword: SCFA

components from the roots of Solanum pseudocapsicum.

The components obtained by hydrodistillation of Solanum pseudocapsicum roots were analyzed by gas chromatography-mass spectrometry. A total of 41 compounds, representing 50% of the oil, were identified. The oil was found to contain (26.8%), terpenoids (7.6%), and aldehydes (5.3%) as the major components. The dominant compounds were hexadecanoic (24.1%), 2-methoxy-3-isopropylpyrazine (2.8%), and 15-methylhexadecanoic (2.1%). Other notable components include beta-elemene and delta-elemene. The high proportion of in this plant could contribute to its medicinal properties.

Keyword: SCFA

Chemical Characterization of Lodoicea maldivica Fruit.

In the present study, we report the attempt to characterize the chemical composition of fruit kernel of Lodoicea maldivica coco nucifera palm (commonly named as \'Coco de mer\') by gas chromatographic method. The analysis was performed by HS-SPME and GC/MS techniques to determine aroma, sterol, and composition profiles in the internal and external pulp of two distinct coconuts. Although no qualitative differences in flavour composition were observed between the two analysed coconuts and the relative two pulp parts, variations in the abundance levels of the prominent compounds have been recorded. The averaged quantity of total phytosterols, resulting from the two analysed \'Coco de mer\' samples, was almost constant in both kernels coconut, being 24.5 μg/g (of dry net matter) for the external, and 26.9 μg/g (of dry net matter) for the internal portion. In both coconuts, the pattern composition was characterized by seven saturated ranged from C14:0 (myristic) to C20:0 (arachidic) and two monounsaturated , the palmitoleic (C16:1, ω7) and the oleic (C18:1, ω9). (C16:0) was the predominant one with an average contribution of about 49.0%, followed by pentadecanoic 16.5%, stearic (C18:0) 11.6%, and myristic (C14:0) 9.9% in all two examined kernel portions.© 2017 Wiley-VHCA AG, Zurich, Switzerland.

Keyword: SCFA

molecular markers of VOO Thermo-oxidation: Effect of heating processes, macronutrients composition, and olive ripeness on the new emitted aldehydic compounds.

Heating operation has been applied to Chétoui extra-virgin olive oils (EVOOs) extracted from fruits with several ripening stages (RS). The studied samples, were subjected to microwave and conventional heating. Results showed that heated VOOs after 2.5\u202fh and 7\u202fmin of conventional and microwave heating, respectively, gave rise to a drastically decrease of LOX products and allowed the detection of toxic new formed aldehydic volatiles (alkanal: nonanal, alkenals: (Z)-2-heptenal and (E)-2-decenal, and alkadienals: (E.E)-2.4-decadienal), which can be used as markers of VOO degradation. Their abundance in the VOO headspaces depends on their boiling points, the rate of their possible degradation to yield other compounds, on the heating processes and on the rate of macronutrients. The emission rate of the new synthesized volatiles during heating processes was mainly attributed to enzymatic oxidation of some . Hexanal, (Z)-2-heptenal, (E)-2-octenal, (E)-2-nonenal, (E,E) and (E,Z)-2,4-decadienal, and (E,E)-2,4-nonadienal, derived from linoleic , and heptanol, octanal, nonanal, decanal, (E) and (Z)-2-decenal, (E)-2-undecenal, and (E,E)-2,4-nonadienal, are emitted after degradation of oleic . During thermo-oxidation, the ECN (LLO, and OLnO), and the ECN (OLO, and PLO\u202f+\u202fSLL) compounds decreased, whereas, the ECN (OOO, and PPO), and the ECN (SOO) compounds increased when temperature and heating time increased. The several variations of the studied biochemical compounds depend to the heating processes. Ripening stage of olive fruits can be used as a tool to monitor the emission rate of the aldehydic volatiles, but cannot be used for a chemometric discrimination.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: SCFA

Abundances of Triacylglycerol Positional Isomers and Enantiomers Comprised of a Dipalmitoylglycerol Backbone and Short- or Medium-chain Fatty Acids in Bovine Milk Fat.

Bovine milk fat (BMF) is composed of triacylglycerols (TAG) rich in (P), oleic (O), and short-chain or medium-chain fatty acids (SCFAs or MCFAs). The composition and binding positions of the fatty acids on the glycerol backbone determine their physical and nutritional properties. SCFAs and MCFAs are known to characteristically bind to the sn-3 position of the TAGs in BMF; however, there are very few non-destructive analyses of TAG enantiomers binding the fatty acids at this position. We previously reported a method to resolve the enantiomers of TAGs, binding both long-chain saturated fatty and unsaturated fatty at the sn-1 and 3 positions, in palm oil, fish oil, and marine mammal oil using chiral HPLC. Here, we further developed a method to resolve several TAG enantiomers containing a dipalmitoyl (PP) glycerol backbone and one SCFA (or MCFA) in BMF. We revealed that the predominant TAG structure in BMF was homochiral, such as 1,2-dipalmitoyl-3-butyroyl-sn-glycerol. This is the first quantitative determination of many TAG enantiomers, which bind to a SCFA or MCFA, in BMF was evaluated simultaneously. Furthermore, the results indicated that the amount ratios of the positional isomers and enantiomers of TAGs consisting of a dipalmitoyl (PP) glycerol backbone and SCFA (or MCFA), resembled the whole TAG structures containing the other diacylglycerol backbones consisting of P, O, myristic , and/or stearic in BMF.

Keyword: SCFA

Lipid metabolism in liver of rat exposed to cadmium.

We investigated the effect of exposition to cadmium (Cd, 15ppm for 8 weeks) through drinking water on liver lipid metabolism in adult male Wistar rats. As compared to metal non-exposed (control) rats, the serum triglycerides, cholesterol and LDL+VLDL cholesterol concentrations increased. This was associated to a decrease of lipoprotein lipase activity in post heparinic plasma. The VLDL secretion from liver was not modified. Cd treatment increased triglycerides and decreased esterified cholesterol contents in liver. The high triglyceride mass was related to the increased glycerol-3-phosphate acyltransferase mRNA expression. In addition, the liver synthesis increased, as determined by an increment of synthetase and isocitrate dehydrogenase activities, and [(14)C]-acetate incorporation into saponifiable lipid fraction. The relative percentage of (16:0) and total saturated were increased compared with control. Hepatic glucose-6-phosphate dehydrogenase, malic dehydrogenase and cholesteryl ester hydrolase activities were unchanged. In liver, the Cd treatment decreased triglyceride and cholesterol in mitochondria, also increased triglyceride in cytosol, and cholesterol and phospholipid contents in nuclei, compared with control. In addition, an increase of nuclei phosphatidylcholine synthesis was observed. Cd exposure alters directly or indirectly the serum lipid content and liver lipid metabolism.

Keyword: SCFA

Central administration of palmitoylethanolamide reduces hyperalgesia in mice via inhibition of NF-kappaB nuclear signalling in dorsal root ganglia.

Despite the clear roles played by peroxisome proliferators-activated receptor alpha (PPAR-alpha) in lipid metabolism, inflammation and feeding, the effects of its activation in the central nervous system (CNS) are largely unknown. Palmitoylethanolamide (PEA), a member of the - ethanolamide family, acts peripherally as an endogenous PPAR-alpha agonist, exerting analgesic and anti-inflammatory effects. Both PPAR-alpha and PEA are present in the CNS, but the specific functions of this lipid and its receptor remain to be clarified. Using the carrageenan-induced paw model of hyperalgesia in mice, we report here that intracerebroventricular administration of PEA (0.1-1 microg) 30 min before carrageenan injection markedly reduced mechanical hyperalgesia up to 24 h following inflammatory insult. This effect was mimicked by GW7647 (1 microg), a synthetic PPAR-alpha agonist. The obligatory role of PPAR-alpha in mediating PEA\'s actions was confirmed by the lack of anti-hyperalgesic effects in mutant mice lacking PPAR-alpha. PEA significantly reduced the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in sciatic nerves and restored carrageenan-induced reductions of PPAR-alpha in the L4-L6 dorsal root ganglia (DRG). To investigate the mechanism by which PEA attenuated hyperalgesia, we evaluated inhibitory kB-alpha (IkB-alpha) degradation and p65 nuclear factor kB (NF-kappaB) activation in DRG. PEA prevented IkB-alpha degradation and p65 NF-kappaB nuclear translocation, confirming the involvement of this transcriptional factor in the control of peripheral hyperalgesia. These results add further support to the broad-spectrum of biological and pharmacological effects induced by PPAR-alpha agonists, suggesting a centrally mediated component for these drugs in controlling inflammatory pain.

Keyword: SCFA

Essential oils from stem and leaves of Angelica urumiensis (Mozaffarian) from Iran.

The composition of essential oils of the leaves and stem of Angelica urumiensis were analysed by chromatography-mass spectrometry. Overall, 58 components were identified on the basis of their mass spectra characteristics and retention indices. Twenty-seven compounds were identified in the oil of the leaves, comprising 94.69% of the total oil, in which alpha-cadinol (20.2%), (14.14%), hexahydrofarnesyl acetone (10.03%), 1-dodecanol (7.55%), linoleic (6.37%) and oleic (5.34%) were the major constituents. Oxygenated sesquiterpenes and were the main groups of compounds with 30.7% and 25.85%, respectively. Fifty compounds, representing 96.35% of the total oil, were identified in the stem oil. (13.37), alpha-cadinol (9.24%), (epi)-alpha-cadinol (5.76%) and delta-cadenine (6.11%) were the major compounds. Sesquiterpenes and oxygenated sesquiterpenes dominated in the oil, comprising 28.03% and 20.9%, respectively. The chemical compounds of the essential oils showed that there are only 22 common compounds between two parts.

Keyword: SCFA

Ethanol contamination leads to ethyl esters in hair samples.

The diagnosis of alcoholism is a topical subject of discussion; in fact, many studies have been published on the determination of biochemical markers useful to this target. ethyl esters (FAEE) are minor metabolites of ethanol, and their usefulness has been demonstrated by their detection in hair using a headspace solid-phase microextraction-gas chromatographic-mass spectrometric technique. Environmental contamination in the analysis of drugs of abuse is a well-known focus of discussion between scientists. In the same way, interference from the surroundings could be hypothesized in FAEE detection. To assess the influence of ethanol contamination, an in vitro experiment was performed, leaving hair in an atmosphere saturated with ethanol vapors for 15 days. The spontaneous production of FAEE was demonstrated by analyzing hair day by day. In fact, we observed a constant increase of ethyl myristate, palmitate, and stearate that reached very high concentrations at the end of the investigation. Although the experiment was managed in a stressed way and could not represent real life, its purpose was to focus the attention of researchers on the problem of hair contamination that can occur, for example, with ethanol-containing cosmetics. Therefore, care in interpretation must be taken into account, especially with such a molecule.

Keyword: SCFA

Quantifying protein- interactions using electrospray ionization mass spectrometry.

The application of the direct electrospray ionization mass spectrometry (ESI-MS) assay to quantify interactions between bovine β-lactoglobulin (Lg) and a series of (FA), CH(3)(CH(2))(x)COOH, where x=6 (caprylic , CpA), 8 (capric , CA), 10 (lauric , LA), 12 (myristic , MA), 14 (, PA) and 16 (stearic , SA), is described. Control ESI-MS binding measurements performed on the Lg-PA interaction revealed that both the protonated and deprotonated gas phase ions of the (Lg + PA) complex are prone to dissociate in the ion source, which leads to artificially small association constants (K (a)). The addition of imidazole, a stabilizing solution additive, at high concentration (10 mM) increased the relative abundance of (Lg + PA) complex measured by ESI-MS in both positive and negative ion modes. The K(a) value measured in negative ion mode and using sampling conditions that minimize in-source dissociation is in good agreement with a value determined using a competitive fluorescence assay. The K (a) values measured by ESI-MS for the Lg interactions with MA and SA are also consistent with values expected based on the fluorescence measurements. However, the K (a) values measured using optimal sampling conditions in positive ion mode are significantly lower than those measured in negative ion mode for all of the FAs investigated. It is concluded that the protonated gaseous ions of the (Lg + FA) complexes are kinetically less stable than the deprotonated ions. In-source dissociation was significant for the complexes of Lg with the shorter FAs (CpA, CA, and LA) in both modes and, in the case of CpA, no binding could be detected by ESI-MS. The affinities of Lg for CpA, CA, and LA determined using the reference ligand ESI-MS assay, a method for quantifying labile protein-ligand complexes that are prone to in-source dissociation, were found to be in good agreement with reported values.© American Society for Mass Spectrometry, 2011

Keyword: SCFA

Expression of genes associated with aroma formation derived from the pathway during peach fruit ripening.

Changes in characteristic aroma volatiles, levels of as aroma precursors, and expression patterns of related genes, including lipoxygenase (LOX), hydroperoxide lyase (HPL), alcohol dehydrogenase (ADH), alcohol acyltransferase (AAT), and desaturase (FAD), were studied in peach ( Prunus persica L. Batsch., cv. Yulu) fruit during postharvest ripening at 20 degrees C. Concentrations of n-hexanal, (E)-2-hexenal, (E)-2-hexenol, and (Z)-3-hexenol decreased, whereas the production of (Z)-3-hexenyl acetate, gamma-hexalactone, gamma-octalactone, gamma-decalactone, and delta-decalactone increased with fruit ripening. Lactones showed a clear pattern concomitant with the climacteric rise in ethylene production, with gamma-decalactone being the principal compound at the late ripening stage. Of the LOX family genes, PpLOX2 and PpLOX3 had relatively high transcript levels initially followed by a decline with fruit ripening, while levels of PpLOX1 and PpLOX4 transcripts were upregulated by accumulated ethylene production. Expression of PpHPL1, PpADH1, PpADH2, and PpADH3 showed similar decreasing patterns during ripening. Expression levels of PpAAT1 showed a rapid increase during the first 2 days of postharvest ripening followed by a gradual decrease. Contents of polyunsaturated linoleic and linolenic increased, and saturated levels tended to decline as the fruit ripened. The increased levels of unsaturated closely paralleled increasing expression of PpFAD1 and PpFAD2. The significance of gene expression changes in relation to aroma production is discussed.

Keyword: SCFA

Associations between hepatic metabolism of propionate and palmitate in liver slices from transition dairy cows.

Multiparous Holstein cows (n=95) were used to evaluate changes in hepatic propionate and palmitate metabolism and liver composition over time during the transition period, along with the relationships of these variables with cumulative increases in nonesterified and β-hydroxybutyrate during the periparturient period. Data from 3 previous experiments were used to address the study objectives, accounting for a total of 95 multiparous Holstein cows. Liver slices from biopsies on d -21, 1, and 21 relative to parturition were used to determine conversion of [1-(14)C]palmitate to CO2 and esterified products (EP) and the conversion of [1-(14)C]propionate to CO2 and glucose. Hepatic glycogen content was highest on d -21 and was 26.9 and 36.5% of prepartum values on d 1 and 21, respectively. Liver triglyceride content was lowest at d -21 and was 271 and 446% of prepartum values on d 1 and 21, respectively. We detected no difference in the capacity for the liver to oxidize [1-(14)C]palmitate to CO2 between d -21 and d 1; however, on d 21, oxidation was 84% of prepartum values. The capacity of the liver to convert [1-(14)C]palmitate to EP was 148 and 139% of prepartum values on d 1 and 21, respectively. The capacity of liver to convert [1-(14)C]propionate to CO2 was 127 and 83% of prepartum values on d 1 and 21, and the capacity of liver to convert [1-(14)C]propionate to glucose was 126 and 85% of prepartum values on d 1 and 21, respectively. Correlation relationships suggest that overall, cows with elevated prepartum liver triglyceride content had elevated triglycerides throughout the transition period along with increased [1-(14)C]palmitate oxidation and conversion to EP and a decreased propensity to convert [1-(14)C]propionate to glucose. Cows with increased [1-(14)C]propionate oxidation had increased conversion of [1-(14)C]propionate to glucose throughout the transition period. Overall, conditions that lead to impairments in metabolism during the transition period appear to be associated with impaired postpartum hepatic propionate metabolism.Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Anaerobic biodegradation of oleic and : evidence of mass transfer limitations caused by long chain accumulation onto the anaerobic sludge.

was the main long chain (LCFA) that accumulated onto the anaerobic sludge when oleic was fed to an EGSB reactor. The conversion between oleic and was linked to the biological activity. When was fed to an EGSB reactor it represented also the main LCFA that accumulated onto the sludge. The way of accumulation was different in the oleic and in the fed reactors. When oleic was fed, the biomass-associated LCFA (83% as ) were mainly adsorbed and entrapped in the sludge that became "encapsulated" by an LCFA layer. However, when was fed, the biomass-associated LCFA (the totality as ) was mainly precipitated in white spots like precipitates in between the sludge, which remained "non-encapsulated." The two sludges were compared in terms of the specific methanogenic activity (SMA) in the presence of acetate, propionate, butyrate, and H(2)CO(2), before and after the mineralization of similar amounts of biomass-associated LCFA (4.6 and 5.2 g COD-LCFA/g of suspended solids (VSS), for the oleic and fed sludge, respectively). The "non-encapsulated," sludge exhibited a considerable initial methanogenic activity on all the tested substrates, with the single exception of butyrate. However, with the "encapsulated" sludge only methane production from ethanol and H(2)/CO(2) was detected, after a lag phase of about 50 h. After mineralization of the biomass-associated LCFA, both sludges exhibited activities of similar order of magnitude in the presence of the same individual substrates and significantly higher than before. The results evidenced that LCFA accumulation onto the sludge can create a physical barrier and hinder the transfer of substrates and products, inducing a delay on the initial methane production. Whatever the mechanism, metabolic or physical, that is behind this inhibition, it is reversible, being eliminated after the depletion of the biomass-associated LCFA.

Keyword: SCFA

Oleic is a potent inhibitor of and cholesterol synthesis in C6 glioma cells.

Glial cells play a pivotal role in brain metabolism and membrane biogenesis. However, the potential regulation of lipogenesis and cholesterologenesis by in glial cells has been barely investigated. Here, we show that physiologically relevant concentrations of various saturated, monounsaturated, and polyunsaturated significantly reduce [1-(14)C]acetate incorporation into and cholesterol in C6 cells. Oleic was the most effective at depressing lipogenesis and cholesterologenesis; a decreased label incorporation into cellular , stearic, and oleic was detected, suggesting that an enzymatic step(s) of de novo biosynthesis was affected. To clarify this issue, the activities of acetyl-coenzyme A carboxylase (ACC) and FAS were determined with an in situ digitonin-permeabilized cell assay after incubation of C6 cells with . ACC activity was strongly reduced ( approximately 80%) by oleic , whereas no significant change in FAS activity was observed. Oleic also reduced the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR). The inhibition of ACC and HMGCR activities is corroborated by the decreases in ACC and HMGCR mRNA abundance and protein levels. The downregulation of ACC and HMGCR activities and expression by oleic could contribute to the reduced lipogenesis and cholesterologenesis.

Keyword: SCFA

Amino and composition of an aqueous extract of Channa striatus (Haruan) that exhibits antinociceptive activity.

1. The present study was performed in order to determine the amino and composition of an aqueous extract of the freshwater fish Channa striatus, obtained by soaking (1:2, w/v) fresh fillets overnight in a chloroform:methanol (2:1, v/v) solvent, to elucidate the mechanism responsible for its antinociceptive activity and to clarify the relationship between the presence of the amino and and the expected activity. 2. The aqueous extract was found to contain all amino with the major amino glycine, alanine, lysine, aspartic and proline making up 35.77 +/- 0.58, 10.19 +/- 1.27, 9.44 +/- 0.56, 8.53 +/- 1.15 and 6.86 +/- 0.78% of the total protein, respectively. 3. In addition, the aqueous extract was found to have a high (C16:0) content, which contributed approximately 35.93 +/- 0.63% to total . The other major in the aqueous extract were oleic (C18:1), stearic (C18:0), linoleic (C18:2) and arachidonic (C20:4), contributing 22.96 +/- 0.40, 15.31 +/- 0.33, 11.45 +/- 0.31 and 7.44 +/- 0.83% of total , respectively. 4. Furthermore, the aqueous extract was demonstrated to possess concentration-dependent antinociceptive activity, as expected, when assessed using the abdominal constriction test in mice. 5. It is concluded that the aqueous extract of C. striatus contains all the important amino , but only some of the important , which are suggested to play a key role in the observed antinociceptive activity of the extract, as well as in the traditionally claimed wound healing properties of the extract.

Keyword: SCFA

Biosynthesis of poly-beta-hydroxyalkanoates by Sphingopyxis chilensis S37 and Wautersia sp. PZK cultured in cellulose pulp mill effluents containing 2,4,6-trichlorophenol.

Poly-beta-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications. Kraft cellulose industry effluents containing 2,4,6-trichlorophenol (10 or 20 microg ml(-1)) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating that these polymers are made by 3-hydroxybutyric and 3-hydroxyhexadecanoic for S37 and PZK strains, respectively. Results demonstrated that strains\' growth and PHA production and composition are not modified in cellulose effluents with or without 2,4,6-TCP (10-20 microg ml(-1)). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value substrates.

Keyword: SCFA

Composition and Constituents of Protaetia brevitarsis Larvae.

A total of 48 different oils were identified form P. brevitarsis larvae by gas chromatography/mass spectrometry (GC/MS). (48.67%) were detected as the major group in P. brevitarsis larvae comprising the largest proportion of the compounds, followed by esters (19.84%), hydrocarbons (18.90%), alcohols (8.37%), miscellaneous (1.71%), aldehydes (1.35%) and terpenes (1.16%). The major constituents were 9-hexadecenoic (16.75%), 6-octadecenoic (14.88%) and n-hexadecanoic (11.06%). The composition of was also determined by GC analysis and 16 were identified. The predominant were oleic (C18:1, 64.24%) followed by (C16:0, 15.89%), palmitoleic (C16:1, 10.43%) and linoleic (C18:2, 4.69%) constituting more than 95% of total . The distinguished characteristic of the profile of P. brevitarsis larvae was the high proportion of unsaturated (80.54% of total ) versus saturated (19.46% of total ). Furthermore, small but significant amounts of linoleic, linolenic and γ-linolenic bestow P. brevitarsis larvae with considerable nutritional value. The novel findings of the present study provide a scientific basis for the comprehensive utilization of the insect as a nutritionally promising food source and a possibility for more effective utilization.

Keyword: SCFA

Long-chain saturated induce pro-inflammatory responses and impact endothelial cell growth.

Saturated (SFAs), significant components of enteral and parenteral formulations, have been linked to cardiovascular complications. However, the effect of SFAs upon vascular inflammation is less clear. Endothelial cells (EC) play an important role in the acute inflammatory responses. We, therefore, evaluated the acute effects of different chain-length SFAs upon EC functions.Endothelial cells were cultured with various SFAs. Growth and cytotoxicity were determined by WST-1 assay. Apoptosis and pro-inflammatory adhesion molecule (ICAM-1) expression was assayed using flow cytometry. Activation of NF-kappaB was analyzed using western blot analysis.Long-chain SFAs (C14:0-C20:0) inhibited EC growth in a chain-length dependent manner. Medium-chain SFAs (C6:0-C12:0) did not significantly affect EC growth. In contrast, the short-chain SFA (C4:0) stimulated cellular growth. Stearic induced significantly more EC apoptosis and necrosis than or myristic . Stearic (>10muM) treatment also significantly increased ICAM-1 expression. Stearic \'s pro-inflammatory response was confirmed by phosphorylation of IkappaB-alpha and NF-kappaB in a dose dependent manner.Long-chain SFAs can induce pro-inflammatory responses and significantly impact growth and viability of EC. Our data suggest that the presence of long-chain SFAs in parenteral formulations may have harmful effects on the vascular system.Copyright 2009 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Keyword: SCFA

Lactobacillus parafarraginis ZH1 producing anti-yeast substances to improve the aerobic stability of silage.

Enhancing the aerobic stability of silage is very important in silage production. The objective of this study was to compare the roles of a new bacterial strain Lactobacillus parafarraginis ZH1 with Lactobacillus plantarum Chikuso-1 (LP) and Lactobacillus\xa0buchneri NCIMB 40788 (LB) in improving the aerobic stability of oat silage and identify the anti-yeast substances produced by them. After ensiled for 45\xa0days, either inoculated silages or control silage were of pH values lower than 4.2. The control and LP inoculated silage had poorer aerobic stability than LB and ZH1 inoculated ones (p\xa0<\xa00.05). ZH1 inoculated silage produced more acetic , benzoic and hexadecanoic than LB inoculated one and had the best aerobic stability (p\xa0<\xa00.05). In MRS medium, strain ZH1 produced more hexadecanoic than LP and LB strains, more benzoic than strain LP (p\xa0<\xa00.05), and the equal amount of benzoic to LB (p\xa0>\xa00.05). In addition, benzoic and hexadecanoic had low minimal inhibitory concentrations to target yeasts in the pure culture. In conclusion, L.\xa0parafarraginis ZH1 had stronger anti-yeast potential and more effectively improved the aerobic stability of silage than other strains used in this study.© 2018 Japanese Society of Animal Science.

Keyword: SCFA

Content and composition of sulfoquinovosyldiacylglycerol in conifer leaves grown in Hokkaido, Japan.

Sulfoquinovosyldiacylglycerol (SQDG) contents in conifer leaves and their (FA) compositions were determined. The SQDG content was 16-36 mg/100 g, and was high in Picea glehnii. and alpha-linolenic were the usually predominant FAs. In Picea, the proportion of alpha-linolenic was low, and those of oleic and linoleic were high. The essential oil residues of Abies sachalinensis leaves were found to be a potential source of SQDG material.

Keyword: SCFA

Comparative analysis of metabolomics signals from melanoma and benign skin: a pilot study.

The analysis of organic compounds (VOC) as biomarkers of cancer is both promising and challenging. In this pilot study, we used an untargeted approach to compare metabolomic signatures of melanoma and matched control non-neoplastic skin from the same patient. VOC from fresh (non-fixed) biopsied tissue were collected using the headspace solid phase micro extraction method (HS SPME) and analyzed by gas chromatography and mass spectrometry (GCMS). We applied the XCMS analysis platform and MetaboAnalyst software to reveal many differentially expressed metabolic features. Our analysis revealed increased levels of lauric (C12:0) and (C16:0) in melanoma. The identity of these compounds was confirmed by comparison with chemical standards. Increased levels of these are likely to be a consequence of up-regulated de novo lipid synthesis, a known characteristic of cancer. Increased oxidative stress is likely to cause an additional increase in lauric . Implementation of this study design on larger number of cases will be necessary for the future metabolomics biomarker discovery applications.

Keyword: SCFA

Effects of feeding whole linseed on ruminal composition and microbial population in goats.

The objective of the present study was to evaluate the effect of feeding different levels of whole linseed, as a source of n-3 polyunsaturated (PUFA), on ruminal composition and microbial population in the goat. Twenty-four crossbred Boer goats were assigned to 3 dietary treatments: L0 (control), L10 and L20 containing 0, 10%, or 20% whole linseed, respectively. The ruminal pH and concentration of total (VFA) were not affected by dietary treatments. The feeding of L10 and L20 diets produced higher (\xa0<\xa00.05) molar proportions of acetate and lower (\xa0<\xa00.05) molar proportions of butyrate and valerate than the L0 diet. Molar proportions of myristic (C14:0) and (C16:0) were lower (\xa0<\xa00.05) in the rumen of goats offered L10 and L20 diets than the control diet. However, stearic (C18:0), vaccenic (C18:1 trans-11), conjugated linoleic (CLA, C18:2 trans-10, cis-12) and α-lenolenic (C18:3 n-3) were higher (\xa0<\xa00.05) in the rumen of goats fed L10 and L20 than L0. Both inclusion levels of linseed in the diet (L10 and L20) reduced the ruminal total bacteria, methanogens, and protozoa compared with L0 (\xa0<\xa00.05). The effect of the dietary treatments on cellulolytic bacteria, varied between the individual species. Both inclusion levels of linseed resulted in a significant decrease (\xa0<\xa00.05) in the population of , and compared with L0, with no significant difference between the groups fed linseed diets. The population of was not affected by the different dietary treatments. It was concluded that inclusion of whole linseed in the diet of goats could increase the concentration of PUFA in the rumen, and decrease the population of , , methanogens and protozoa in rumen liquid of goats.

Keyword: SCFA

The influence of the position of palmitate in infant formula triacylglycerols on health outcomes.

The purpose of this review is to discuss recent studies reporting on the influence of the position of in triacylglycerols in infant formula and relevant animal studies. Earlier experiments in rodents show that a diet with a higher proportion of palmitate at the sn-2 position of triacylglycerols improves dietary fat and calcium absorption compared with a diet with a lower sn-2 palmitate content. A high-sn-2 palmitate diet increased fecal short-chain fatty acids, reduced gut inflammation in a colitis model, and altered tissue endocannabinoid concentrations in laboratory rodents. Recent studies in infants confirm that formula with a high sn-2 palmitate content reduces stool fat, , fat soaps, palmitate soaps, and calcium compared with formula with a low sn-2 palmitate content. These effects have been associated with improved bone strength, increased fecal bifidobacteria, and reduced crying in infants. In some studies, findings with formula high in sn-2 palmitate match those seen in breast-fed infants. However, in many studies, high sn-2 palmitate formula remains inferior to breast-feeding. It is concluded that infant formula high in sn-2 palmitate is superior to formula with low sn-2 palmitate but does not fully match human breast milk. Recent studies showing altered gut (human infants) and tissue endocannabinoids (rodent model) suggest the potential for marked physiological impact of high sn-2 palmitate that needs to be explored further in human trials.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: SCFA

Biochemical evaluation of borage (Borago officinalis) rosette leaves through their essential oil and composition.

Borago officinalis rosette leaves were sampled in the region of Amdoun (Tunisia) during different stages of their development. Essential oil contents varied from 0.01% to 0.13% respectively in young and adult leaves. Twenty three compounds were identified. Hydrocarbons, mainly represented by nonadecane (29.8%), tetracosane (11.3%) and heptacosane (4.7%), constituted the major class in the young leaves (45.8%), followed by aldehydes (22.4%). The percentages of these two classes decreased to reach respectively 15% and 8.1% in adult leaves in favour of alcohols (57.9%) where cis-3-hexenol (29.6%) and hexanol (14.5%) were the main compounds. Total amounts increased from 5.03 mg/g DW in young leaves to 32.23 mg/g DW in adult ones. The predominant were alpha-linolenic (C18:3 n-3), stearidonic (C18:4 n-3), gamma-linolenic (C18:3 n-6), (C16: 0) and linoleic (C18:2 n-6) .

Keyword: SCFA

Comparision of the components of unripe and ripe Japanese apricot (Prunus mume Sieb. et Zucc.).

The composition of the components from unripe (I) and ripe (II) Japanese apricot (Prunus mume Sieb. et Zucc.) have been investigated. Seventy-six components were identified, 25 for the first time as constituents of Japanese apricot. The main components were benzaldehyde (I, 59.16%; II, 1.81%), isolongifololyl acetate (II, 19.21%), (I, trace; 10.22%), linalool (I, 9.93%; II, 7.34%), and butyl acetate (II, 8.30%). Unripe Japanese apricot have a green colour due to the main components being aldehydes. On the contrary, ripe Japanese apricot had a fruity note due to the increase of the ratio of esters during ripening.

Keyword: SCFA

composition and compounds of caviar from farmed white sturgeon (Acipenser transmontanus).

The present study was conducted to characterize caviar obtained from farmed white sturgeons (Acipenser transmontanus) subjected to different dietary treatments. Twenty caviar samples from fish fed two experimental diets containing different dietary lipid sources have been analysed for chemical composition, and flavour compounds. make up of caviar was only minimally influenced by dietary composition. Irrespective of dietary treatments, (16:0) and oleic (OA, 18:1 n-9) were the most abundant followed by docosahexaenoic (DHA, 22:6 n-3) and eicopentaenoic (EPA, 20:5 n-3). Thirty-three compounds were isolated using simultaneous distillation-extraction (SDE) and identified by GC-MS. The largest group of volatiles were represented by aldehydes with 20 compounds, representing the 60% of the total volatiles. n-Alkanals, 2-alkenals and 2,4-alkadienals are largely the main responsible for a wide range of flavours in caviar from farmed white surgeon.

Keyword: SCFA

Evaluation of the Key Odorants in Oils from Tubers of Apios americana Medikus.

This study was investigated the chemical composition of oils and aroma evaluation from the tubers of Apios americana Medikus. Theses oils were obtained by the hydrodistillation (HD) and the solvent-assisted flavor evaporation (SAFE) methods. These oils were analyzed by Gas chromatography (GC), GC-mass spectrometry (GC-MS), GC-olfactometry (GC-O), aroma extract dilution analysis (AEDA) and odor activity values (OAV) for the first time. The major compounds in the HD oil were (36.5%), linoleic (10.5%) and nonadecanol (5.7%). Meanwhile, in the SAFE oil, the major compounds were 4-hydroxy-4-methyl-2-pentanone (34.2%), hexanal (11.0%) and hexanol (7.9%). Through aroma evaluation, 20 (HD) and 14 (SAFE) aroma-active compounds were identified by GC-O. As a result, the most intense aroma-active compounds in both extraction methods were 1-octen-3-ol and hexanal, both of which showed high odor activity values (OAV).

Keyword: SCFA

[Fibrate-induced changes in the serum lipid contents of individual C16 and C18 in patients with hyperlipidemia].

Keyword: SCFA

Identification of mosquito biting deterrent constituents from the Indian folk remedy plant Jatropha curcas.

An investigation of the Indian folk remedy plant Jatropha curcas L., was performed to identify the constituents responsible for the mosquito biting deterrent activity of the oil. J. curcas seed oil is burned in oil lamps in India and parts of Africa to repel biting insects, primarily mosquitoes. The seed oil was thoroughly analyzed by 1H NMR, 13C NMR, high-performance liquid chromatography-refractive index, and gas chromatography-flame ionization detection to identify the constituents in the oil. Identified constituents, both free and triglycerides, were evaluated for activity in Aedes aegypti (L.) (Diptera: Culicidae) biting deterrent assays. Furthermore, an oil condensation trap was used to demonstrate that free or triglycerides are partially volatilized during the combustion process. These compounds were found to be responsible for the biting deterrency of the burned oil. Specifically, oleic, , linoleic, and stearic were all active at 25 nmol/cm2 above that of solvent control in Ae. aegypti biting deterrent assays. Oleic, , and linoleic were all more active than stearic in the same bioassay. Evaluation of the triglycerides containing each of these revealed that tripalmitin, tristearin, trilinolein, and triolein all demonstrated significant activity above a solvent control at 10 microg/cm2, whereas tripalmitin was the most active. Due to literature reports suggesting larvicidal activity of the oil, J. curcas seed oil and its free constituents also were evaluated against 1-d-old Ae. aegypti larvae up to 500 ppm. Oleic was the only having larvicidal activity against 1-d-old Ae. aegypti larvae, with an LD50 of 47.9 ppm.

Keyword: SCFA

Characterization of French Coriander Oil as Source of Petroselinic .

Coriander vegetable oil was extracted from fruits of French origin in a 23% yield. The oil was of good quality, with a low amount of free (1.8%) and a concurrently high amount of triacylglycerols (98%). It is a rich source of petroselinic (C18:1n-12), an important renewable building block, making up 73% of all , with also significant amounts of linoleic (14%), oleic (6%), and (3%). The oil was characterized by a high unsaponifiable fraction, comprising a substantial amount of phytosterols (6.70 g/kg). The main sterol markers were β-sitosterol (35% of total sterols), stigmasterol (24%), and Δ⁷-stigmastenol (18%). Squalene was detected at an amount of 0.2 g/kg. A considerable amount of tocols were identified (500 mg/kg) and consisted mainly of tocotrienols, with γ-tocotrienol as the major compound. The phospholipid content was low at 0.3%, of which the main phospholipid classes were phosphatidic (33%), phosphatidylcholine (25%), phosphatidylinositol (17%), and phosphatidylethanolamine (17%). About 50% of all phospholipids were non-hydratable. The β-carotene content was low at 10 mg/kg, while a significant amount of chlorophyll was detected at about 11 mg/kg. An iron content of 1.4 mg/kg was determined through element analysis of the vegetable oil. The influence of fruit origin on the vegetable oil composition was shown to be very important, particularly in terms of the phospholipids, sterols, and tocols composition.

Keyword: SCFA

Comparison of the compositional, microbiological, biochemical and profile characteristics of three Italian PDO fermented sausages.

Three Italian PDO fermented sausages, Varzi, Brianza and Piacentino, were compared for compositional, microbiological, biochemical and profile characteristics. Mean values for the gross composition varied especially due to moisture, fat, total protein and nitrate concentration which reflected differences in the ingredients and some technological parameters. Cell numbers of the major microbial groups were almost similar among sausages. The major differences were found for Brochothrix thermosphacta, enterococci and moulds. Apart from their use as starters, Lactobacillus sakei and Lactobacillus curvatus were the dominant lactic bacteria and, as well as Staphylococcus xylosus, dominated the population of coagulase-negative staphylococci. Sausages differed for the hydrolysis of myofibrillar proteins and secondary proteolysis. Varzi, the sausage subjected to prolonged fermentation at 23-25°C for 10 days before ripening, showed the highest degree of secondary proteolysis. Varzi and Brianza, the two fermented sausages manufactured by using microbial starters, showed the highest concentration and similar profiles of free amino . The peptidase activities contained in the aqueous extracts agreed with the above findings. A total of 52 components, mainly alcohols, aldehydes and terpenes, were identified by solid-phase micro-extraction coupled with gas chromatography-mass spectrometry analysis. The profiles of the three Italian PDO fermented sausages differed in part and, except for terpenes, the highest levels of the other chemical classes were found in Varzi and Brianza sausages. The composition of free of the three Italian PDO sausages was rather similar. Monounsaturated (MUFA) were found at the highest relative percentage followed by saturated (SFA) and polyunsaturated (PUFA) . Oleic, , linoleic and stearic were the main free found in all fermented sausages.

Keyword: SCFA

Milk organic compounds and profile in cows fed timothy as hay, pasture, or silage.

Nutrient composition and organoleptic properties of milk can be influenced by cow diets. The objective of this study was to evaluate the forage type effects on organic compounds, (FA) profile, and organoleptic properties of milk. Timothy grass was fed as hay, pasture, or silage during a period of 27 d to a group of 21 cows in a complete block design based on days in milk. Each cow also received 7.2 kg/d of a concentrate mix to meet their nutrient requirements. Forage dry matter intake averaged 13.9 kg/d and was not different among treatments. Milk yield was higher for cows fed pasture, intermediate for cows fed silage, and lowest for cows fed hay. However, milk fat content was higher for cows fed hay and silage, compared with cows fed pasture. As a result, fat-corrected milk and fat yield were not different among treatments. Increasing the supply of dietary cis-9,cis-12 18:2 (linoleic ) and cis-9,cis-12,cis-15 18:3 (α-linolenic ) when feeding pasture enhanced the concentration of these 2 essential FA in milk fat compared with feeding hay or silage. Moreover, the ratio of 16:0 () to cis-9 18:1 (oleic ), which is closely related to the melting properties of milk fat, was lower in milk from cows on pasture than in milk from cows fed hay or silage. Cows fed hay produced milk with higher levels of several free FA and γ-lactones, but less pentanal and 1-pentanol. More dimethyl sulfone and toluene were found in milk of cows on pasture. Cows fed silage produced milk with higher levels of acetone, 2-butanone, and α-pinene. Results from a sensory evaluation showed that panelists could not detect a difference in flavor between milk from cows fed hay compared with silage. However, a significant number of assessors perceived a difference between milk from cows fed hay compared with milk from cows fed pasture. In a sensory ranking test, the percentage of assessors ranking for the intensity of total (raw milk, fresh milk, and farm milk), sweet (empyreumatic, vanilla, caramel, and sugar), and grassy (grass, leafy vegetable, and plant) flavors was higher for milk from cows fed pasture compared with hay and silage. Using timothy hay, pasture, or silage harvested at a similar stage of development, the current study shows that the taste of milk is affected by the forage type fed to cows. More research is, however, needed to establish a link between the sensory attributes of milk and the observed changes in organic compounds and FA profile.Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Changes on physico-chemical, textural, lipolysis and compounds during the manufacture of dry-cured foal "cecina".

The changes in the physico-chemical and textural properties, lipolysis and compounds during the manufacture of dry-cured foal "cecina" were studied. The pH increased during the last stages of processing but gradually declined over the curing period. TBARS values, hardness and chewiness increased with processing time from 0.14, 2.74 and 0.83 to 3.49 mg malonaldehyde/kg, 20.33 kg and 5.05 kg∗mm, respectively. Ripening time also affected the colour parameters: lightness (L*), redness (a*) and yellowness (b*) (P<0.001). The total average content of free (FFA) increased significantly from 433.7 mg/100 g of fat in the raw pieces to 2655.5 mg/100 g of fat at the end of the drying-ripening stage. The main FFA at the end of the manufacturing process was (C16:0), followed by oleic (C18:1cis9), stearic (C18:0) and linoleic (C18:2n-6). A total of fifty five compounds were identified during the manufacture of dry-cured foal "cecina", including esters, aldehydes, aliphatic hydrocarbons, branched hydrocarbons, alcohols, aromatic hydrocarbons, furans, ketones. Aldehydes reached their maximum level at the end of the post-salting stage. In the final product, esters became the dominant chemical compounds.© 2013.

Keyword: SCFA

Sleep symptoms associated with intake of specific dietary nutrients.

Sleep symptoms are associated with weight gain and cardiometabolic disease. The potential role of diet has been largely unexplored. Data from the 2007-2008 National Health and Nutrition Examination Survey (NHANES) were used (n\xa0=\xa04552) to determine which nutrients were associated with sleep symptoms in a nationally representative sample. Survey items assessed difficulty falling asleep, sleep maintenance difficulties, non-restorative sleep and daytime sleepiness. Analyses were adjusted for energy intake, other dietary factors, exercise, body mass index (BMI) and sociodemographics. Population-weighted, logistic regression, with backwards-stepwise selection, examined which nutrients were associated with sleep symptoms. Odds ratios (ORs) reflect the difference in odds of sleep symptoms associated with a doubling in nutrient. Nutrients that were associated independently with difficulty falling asleep included (in order): alpha-carotene (OR\xa0=\xa00.96), selenium (OR\xa0=\xa00.80), dodecanoic (OR\xa0=\xa00.91), calcium (OR\xa0=\xa00.83) and hexadecanoic (OR\xa0=\xa01.10). Nutrients that were associated independently with sleep maintenance difficulties included: salt (OR\xa0=\xa01.19), butanoic (0.81), carbohydrate (OR\xa0=\xa00.71), dodecanoic (OR\xa0=\xa00.90), vitamin D (OR\xa0=\xa00.84), lycopene (OR\xa0=\xa00.98), hexanoic (OR\xa0=\xa01.25) and moisture (OR\xa0=\xa01.27). Nutrients that were associated independently with non-restorative sleep included butanoic (OR\xa0=\xa01.09), calcium (OR\xa0=\xa00.81), vitamin C (OR\xa0=\xa00.92), water (OR\xa0=\xa00.98), moisture (OR\xa0=\xa01.41) and cholesterol (OR\xa0=\xa01.10). Nutrients that were associated independently with sleepiness included: moisture (OR\xa0=\xa01.20), theobromine (OR\xa0=\xa01.04), potassium (OR\xa0=\xa00.70) and water (OR\xa0=\xa00.97). These results suggest novel associations between sleep symptoms and diet/metabolism, potentially explaining associations between sleep and cardiometabolic diseases.© 2013 European Sleep Research Society.

Keyword: SCFA

Metabolite analysis of endophytic fungi from cultivars of Zingiber officinale Rosc. identifies myriad of bioactive compounds including tyrosol.

Endophytic fungi associated with rhizomes of four cultivars of Zingiber officinale were identified by molecular and morphological methods and evaluated for their activity against soft rot pathogen Pythium myriotylum and clinical pathogens. The bioactive metabolites produced by these isolates were identified by GC-MS analysis of the fungal crude extracts. Understanding of the metabolites produced by endophytes is also important in the context of raw consumption of ginger as medicine and spice. A total of fifteen isolates were identified from the four varieties studied. The various genera identified were Acremonium sp., Gliocladiopsis sp., Fusarium sp., Colletotrichum sp., Aspergillus sp., Phlebia sp., Earliella sp., and Pseudolagarobasidium sp. The endophytic community was unique to each variety, which could be due to the varying host genotype. Fungi from phylum Basidiomycota were identified for the first time from ginger. Seven isolates showed activity against Pythium, while only two showed antibacterial activity. The bioactive metabolites identified in the fungal crude extracts include tyrosol, benzene acetic , ergone, dehydromevalonic lactone, N-aminopyrrolidine, and many bioactive and their derivatives which included linoleic , oleic , myristic , n-hexadecanoic , methyl ester, and methyl linoleate. The presence of these varying bioactive endophytic fungi may be one of the reasons for the differences in the performance of the different ginger varieties.

Keyword: SCFA

Essential-oil and - composition, and antioxidant activity of extracts of Ficaria kochii.

The essential-oil and - composition of the aerial parts of Ficaria kochii (Ledeb.) Iranshahr & Rech.f. native to Iran, and the antioxidant activity of various extracts of this plant were examined. The study by GC-FID and GC/MS analysis of the essential oil resulted in the identification of 61 compounds, representing 86.01% of the total oil composition. Phytol (10.49%), farnesol (7.72%), methyl linoleate (5.57%), and α-farnesene (4.96%) were the main components. The - composition of the aerial parts of F. kochii was also analyzed by GC/MS. The major components were (25.9%), linolenic (25.3%), and linoleic (17.5%). Polyunsaturated (PUFAs) were found in higher amounts than saturated . The possible antioxidant activity of various extracts (prepared by using solvents with different polarity) of the F. kochii aerial parts was evaluated by screening for their 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity, Fe(III) -reducing power, total antioxidant activity, and inhibitory activity in the linoleic -peroxidation system. H(2) O proved to be the most efficient solvent for the extraction of antioxidants, as the H(2) O extract contained the highest amount of phenolic compounds (2.78±0.23 GAE/g dry matter) and also exhibited the strongest antioxidant capacity in all the assays used. The results of the present investigation demonstrated that the aerial parts of F. kochii can be used as natural and safe nutrition supplement in place of synthetic ones.Copyright © 2012 Verlag Helvetica Chimica Acta AG, Zürich.

Keyword: SCFA

Physicochemical evaluation of sheep milk yogurts containing different levels of inulin.

The present study aimed to evaluate the physicochemical parameters of sheep milk yogurt smoothies (SMY) containing inulin at different levels (0, 2, 4, and 6%). Titratable acidity and pH, yogurt bacteria counts, profile, and healthy lipid indices were evaluated during 28 d of refrigerated storage. As expected for yogurts, Streptococcus thermophilus counts decreased 1 to 3 log cycles and Lactobacillus delbrueckii ssp. bulgaricus counts decreased 1 to 2 cycles from d 1 to 28. The protective effect of inulin on bacteria survival and viability in the food matrix was not verified in the prebiotic SMY during storage, regardless of inulin level. Although lower post-acidification was observed in prebiotic SMY due to inulin addition, no changes were verified in short-chain () or polyunsaturated (PUFA). In contrast, an increase in medium- and long-chain (MCFA and LCFA) and monounsaturated (MUFA) was observed during storage in all SMY. The most significant levels of in SMY were oleic , followed by and myristic . A high positive correlation between conjugated linoleic (CLA) and oleic (r=0.978) was observed. The cis-9,trans-11 CLA isomer represented approximately 78% of total PUFA and 2% of total , whereas α-linoleic comprised about 22% PUFA and 1% of total in SMY. The changes during storage were associated with the metabolic activity of the starter bacteria, especially for oleic and cis-9,trans-11 CLA isomer. Thus, the SMY represented a great source of these compounds. We observed that inulin levels did not affect . A nonsignificant decrease in atherogenic index was observed during storage in all SMY, and a positive correlation (r=0.973) was found between atherogenic index and thrombogenic index of SMY. High correlations were observed between lauric and myristic and saturated (r=0.907 and r=0.894, respectively), providing evidence of their atherogenic and thrombogenic potential. A negative correlation was observed between stearic and atherogenic index (r=-0.612) and between oleic and atherogenic index. Sheep milk yogurt could be characterized as a food with low atherogenic and thrombogenic risk because of its healthy lipid composition. Therefore, addition of inulin to SMY could be a good option to improve functionality of this food matrix for dairy companies wishing to enter the functional food market.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: SCFA

Safety and efficacy of Sclerocarya birrea (A.Rich.) Hochst (Marula) oil: A clinical perspective.

Sclerocarya birrea (A.Rich.) Hochst (Marula) nut oil is a popular ingredient in cosmetics such as skin lotions, lipsticks and foundations. The demand for this African oil increased tremendously such that in 2008 almost US$ 20 million was spent on Marula oil for cosmetic product manufacturing. The ethnobotanical literature states that the Zulu people in South Africa amongst others applied the oil to maintain a healthy skin. Scientific studies to support the traditional use as well as the inclusion of Marula oil in cosmetic products is lacking. This study evaluated the irritancy potential (safety), the moisturising and hydrating effects as well occlusivity properties (efficacy) of Marula oil after topical application. In addition, the Marula oil used in this study was comprehensively characterised using two-dimensional gas chromatography coupled to mass spectrometry.Quantification of the fatty methyl esters (FAMEs) was done using a LECO Pegasus 4D GC × GC-MS. To determine the safety and efficacy of Marula oil healthy caucasian adult female volunteers (n = 20) who complied with the inclusion and exclusion criteria for the irritancy patch, moisture efficacy, hydrating and occlusivity tests were recruited for each study. A 2 × magnifying lamp (visual observation), Chromameter®, Aquaflux® and Corneometer® instruments were used to evaluate and monitor the irritancy level, skin , transepidermal water loss, hydrating and occlusive effects of topically applied Marula oil.The GC × GC-MS analysis identified several saturated as well as unsaturated fatty acids. Oleic was the major fatty constituting 69.0% of the oil followed by (15.3%), linoleic (9.2%), palmitoleic (4.1%) and stearic (1.5%). The clinical study revealed that Marula oil is non-irritant (p < 0.001), with moisturising and hydrating properties (p < 0.001) when applied to a lipid-dry (xerosis) skin. Additionally the oil exhibited occlusive effects (p < 0.001) when applied to normal skin. These findings may be linked to the absorption of the oil into the skin due to the high percentage of oleic and the presence of which are known to disturb the stratum corneum intercellular lipids. These fatty acids present in Marula oil are very similar to those present in the epidermis, and can be considered biomimetic.Marula oil rich in fatty acids exhibits moisturising, hydrating and occlusive properties. As the oil is non-irritating and provides a moisturising effect with moderate prevention of transepidermal water loss, average moisture retention properties and noteworthy occlusive effects, its inclusion in cosmetic products based on its traditional use may be justified depending on the application.Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Keyword: barrier function

-Induced Podocyte Apoptosis via the Reactive Oxygen Species-Dependent Mitochondrial Pathway.

Chronic kidney disease (CKD) is often accompanied by hyperlipidemia, which accelerates progression of the disease. Podocyte injury can lead to dysfunction of the glomerular filtration , which is associated with proteinuria, a risk marker for the progression of CKD. Our previous studies demonstrated that (PA) can induce podocyte apoptosis; however, the underlying mechanisms are unclear. In the present study, we investigated the specific molecular mechanisms of PA-induced apoptosis in cultured podocytes.We cultured mouse podocytes and treated them with PA. Then, cell viability was measured using the Cell Counting Kit-8 colorimetric assay, lipid uptake was assessed by Oil Red O staining and boron-dipyrromethene staining, apoptosis was measured by flow cytometry, mitochondrial injury was assessed by JC-1 staining and transmission electron microscopy, and mitochondrial production of reactive oxygen species (ROS) was evaluated by fluorescence microscopy using the MitoSOX Red reagent. The effects of PA on the mitochondria-mediated caspase activation pathway were investigated by examining the expression of caspase-8, cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), B-cell lymphoma 2 (Bcl-2), Bax, Bid, cytochrome c, and Fas-associated protein with death domain (FADD) using western blotting. The translocation of Bax and cytochrome c were detected by immunofluorescence.PA treatment significantly increased lipid accumulation and induced podocyte apoptosis. We investigated whether the two primary apoptosis signaling pathways (death receptor-mediated pathway and mitochondria-mediated pathway) were involved in the execution of PA-induced podocyte apoptosis, and found that the levels of FADD, caspase-8, and Bid did not significantly change during this process. Meanwhile, PA treatment induced an increase in Bax protein expression and a decrease in Bcl-2 protein expression, with Bax translocation to the mitochondria. Furthermore, PA treatment induced mitochondrial impairment, and triggered the release of cytochrome c from the mitochondria to cytosol, with a concomitant dose-dependent increase in the levels of cleaved caspase-9, cleaved caspase-3, and PARP. Meanwhile, PA treatment increased mitochondrial production of ROS, and the mitochondria-targeted antioxidant mitoTEMPO significantly ameliorated PA-induced podocyte apoptosis.Our findings indicated that PA induced caspase-dependent podocyte apoptosis through the mitochondrial pathway, and mitochondrial ROS production participated in this process, thus potentially contributing to podocyte injury.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: barrier function

P38 MAPK Pharmacological Inhibitor SB203580 Alleviates Total Parenteral Nutrition-Induced Loss of Intestinal but Promotes Hepatocyte Lipoapoptosis.

Our previous studies have provided evidence that p38 mitogen-activated protein kinase (MAPK) is involved in total parenteral nutrition (TPN)-associated complications, but its exact effects and mechanisms have not been fully understood. This study aimed to evaluate the roles of p38 MAPK inhibitor SB203580 in the TPN-induced loss of intestinal and liver disease.A rodent model of TPN was used to analyze the roles of SB203580 in TPN-associated complications.Intestinal was evaluated by transepithelial electrical resistance (TER) and paracellular permeability in Caco-2 cells. The (PA) was used to induce hepatic lipoapoptosis in vitro. The lipoapoptosis was detected using Caspase-3/7 and lipid staining.In the present study, we showed that SB203580 treatment significantly suppressed TPN-mediated intestinal permeability in rats. SB203580 treatment significantly inhibited IL-1β-induced an increase in tight junction permeability of Caco-2 cells via repressing the p38/ATF-2 signaling. Unexpectedly, SB203580 treatment enhanced hepatic lipoapoptosis in the model of TPN. (PA)-induced hepatic lipoapoptosis in human liver cells was significantly augmented by the SB203580 treatment.We demonstrate that the p38 MAPK inhibitor SB203508 ameliorates intestinal but promotes hepatic lipoapoptosis in model of TPN.© 2017 The Author(s)Published by S. Karger AG, Basel.

Keyword: barrier function

Chain Length of Free Fatty Acids Influences the Phase Behavior of Stratum Corneum Model Membranes.

The skin, the largest organ of the human body, forms a flexible interface between our internal and external environment that protects our organism from exogenous compounds as well as excessive water loss. The stratum corneum (SC), the outermost layer of mammal epidermis, is mainly responsible for the skin impermeability. The SC is formed by corneocytes embedded in a lipid matrix, which is mostly constituted of ceramides (Cer), free fatty acids (FFA), and cholesterol (Chol), organized in two coexisting crystalline lamellar phases. This arrangement of lipids is crucial to skin . The aim of this paper is to determine the impact of FFA chain length on the phase behavior of SC model lipid membranes using solid-state deuterium NMR and IR spectroscopy. We studied ternary mixtures of N-lignoceroyl-d-erythro-sphingosine (Cer24), cholesterol, and (FFA16) or lignoceric (FFA24) in an equimolar ratio. This proportion replicates the lipid composition found in the SC lipid matrix. Our studies revealed that the phase behavior of Cer24/FFA/Chol ternary mixtures is strongly affected by the length of the FFA. We found the formation of phase-separated crystalline lipid domains when using whereas the use of lignoceric results in a more homogeneous mixture. In addition, it was observed that mixtures with lignoceric form a gel phase, a very unusual feature for SC model mixtures.

Keyword: barrier function

Calcium Uptake via Mitochondrial Uniporter Contributes to -Induced Apoptosis in Mouse Podocytes.

Podocytes are component cells of the glomerular filtration , and their loss by apoptosis is the main cause of proteinuria that leads to diabetic nephropathy (DN). Therefore, insights into podocyte apoptosis mechanism would allow a better understanding of DN pathogenesis and thus help develop adequate therapeutic strategies. Here, we investigated the molecular mechanism of -inhibited cell death in mouse podocytes, and found that increased cell death in a dose- and time-dependent manner. induces apoptosis in podocytes through upregulation of cytosolic and mitochondrial Ca , mitochondrial membrane potential (MMP), cytochrome c release, and depletion of endoplasmic reticulum (ER) Ca . The intracellular calcium chelator, 1,2-bis (2-aminophenoxy) ethane-N,N,N, N\'-tetraacetic tetrakis acetoxymethyl ester (BAPTA-AM), partially prevented this upregulation whereas 2-aminoethoxydiphenyl borate (2-APB), an inositol 1,4,5-triphosphate receptor (IP3R) inhibitor; dantrolene, a ryanodine receptor (RyR) inhibitor; and 4,4\'-diisothiocyanatostibene-2,2\'-disulfonic (DIDS), an anion exchange inhibitor, had no effect. Interestingly, ruthenium red and Ru360, both inhibitors of the mitochondrial Ca uniporter (MCU), blocked -induced mitochondrial Ca elevation, cytochrome c release from mitochondria to cytosol, and apoptosis. siRNA to MCU markedly reduced -induced apoptosis. These data indicate that Ca uptake via mitochondrial uniporter contributes to -induced apoptosis in mouse podocytes. J. Cell. Biochem. 118: 2809-2818, 2017.© 2017 Wiley Periodicals, Inc.

Keyword: barrier function

Branched Fatty Esters of Hydroxy Fatty Acids (FAHFAs) Protect against Colitis by Regulating Gut Innate and Adaptive Immune Responses.

We recently discovered a structurally novel class of endogenous lipids, branched esters of hydroxy stearic acids (PAHSAs), with beneficial metabolic and anti-inflammatory effects. We tested whether PAHSAs protect against colitis, which is a chronic inflammatory disease driven predominantly by defects in the innate mucosal and adaptive immune system. There is an unmet clinical need for safe and well tolerated oral therapeutics with direct anti-inflammatory effects. Wild-type mice were pretreated orally with vehicle or 5-PAHSA (10 mg/kg) and 9-PAHSA (5 mg/kg) once daily for 3 days, followed by 10 days of either 0% or 2% dextran sulfate sodium water with continued vehicle or PAHSA treatment. The colon was collected for histopathology, gene expression, and flow cytometry. Intestinal crypt fractions were prepared for ex vivo bactericidal assays. Bone marrow-derived dendritic cells pretreated with vehicle or PAHSA and splenic CD4 T cells from syngeneic mice were co-cultured to assess antigen presentation and T cell activation in response to LPS. PAHSA treatment prevented weight loss, improved colitis scores (stool consistency, hematochezia, and mouse appearance), and augmented intestinal crypt Paneth cell bactericidal potency via a mechanism that may involve GPR120. In vitro, PAHSAs attenuated dendritic cell activation and subsequent T cell proliferation and Th1 polarization. The anti-inflammatory effects of PAHSAs in vivo resulted in reduced colonic T cell activation and pro-inflammatory cytokine and chemokine expression. These anti-inflammatory effects appear to be partially GPR120-dependent. We conclude that PAHSA treatment regulates innate and adaptive immune responses to prevent mucosal damage and protect against colitis. Thus, PAHSAs may be a novel treatment for colitis and related inflammation-driven diseases.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: barrier function

Maternal metabolic stress may affect oviduct gatekeeper .

We hypothesized that elevated non-esterified fatty acids (NEFA) modify bovine oviduct epithelial cell (BOEC) metabolism and . Hereto, BOECs were studied in a polarized system with 24-h treatments at Day 9: (1) control (0\u2009µM NEFA\u2009+\u20090% EtOH), (2) solvent control (0\u2009µM NEFA\u2009+\u20090.45% EtOH), (3) basal NEFA (720\u2009µM NEFA\u2009+\u20090.45% EtOH in the basal compartment) and (4) apical NEFA (720\u2009µM NEFA\u2009+\u20090.45% EtOH in the apical compartment). FITC-albumin was used for monolayer permeability assessment and related to transepithelial electric resistance (TER). Fatty (FA), glucose, lactate and pyruvate concentrations were measured in spent medium. Intracellular lipid droplets (LD) and FA uptake were studied using Bodipy 493/503 and immunolabelling of FA transporters (FAT/CD36, FABP3 and CAV1). BOEC-mRNA was retrieved for qRT-PCR. Results revealed that apical NEFA reduced relative TER increase (46.85%) during treatment and increased FITC-albumin flux (27.59%) compared to other treatments. In basal NEFA, FAs were transferred to the apical compartment as free FAs: mostly and oleic increased respectively 56.0 and 33.5% of initial FA concentrations. Apical NEFA allowed no FA transfer, but induced LD accumulation and upregulated FA transporter expression (↑CD36, ↑FABP3 and ↑CAV1). Gene expression in apical NEFA indicated increased anti-apoptotic (↑) and anti-oxidative (↑) capacity, upregulated lipid metabolism (↑ ↑ and ↓) and FA uptake (↑). All treatments had similar carbohydrate metabolism and oviduct -specific gene expression (, and ). Overall, elevated NEFAs affected BOEC metabolism and differently depending on NEFA exposure side. Data substantiate the concept of the oviduct as a gatekeeper that may actively alter early embryonic developmental conditions.© 2017 Society for Reproduction and Fertility.

Keyword: barrier function

Molecular Interaction between Intercellular Lipids in the Stratum Corneum and l-Menthol, as Analyzed by Synchrotron X-Ray Diffraction.

l-Menthol increases drug partitioning on the surface of skin, diffusion of drugs in the skin, and lipid fluidity in the stratum corneum and alters the rigidly arranged lipid structure of intercellular lipids. However, l-menthol is a solid at room temperature, and it is difficult to determine the effects of l-menthol alone. In this study, we vaporized l-menthol in order to avoid the effects of solvents. The vaporized l-menthol was applied to the stratum corneum or lipid models comprising composed of ceramides (CER) [EOS], the longest lipid acyl chain of the ceramides in the stratum corneum lipids that is associated with the of the skin; CER [NS], the shorter lipid acyl chain of the ceramides, and the most components in the stratum corneum of the intercellular lipids that is associated with water retention in the intercellular lipid structure of the stratum corneum; cholesterol; and . Synchrotron X-ray diffraction, differential scanning calorimetry, and attenuated total reflection Fourier transform infrared spectroscopy analyses revealed that the lipid models were composed of hexagonal packing and orthorhombic packing structures of different lamellar periods. Taken together, our results revealed that l-menthol strongly affected the lipid model composed of CER [EOS]. Therefore, l-menthol facilitated the permeation of drugs through the skin by liquid crystallization of the longer lamellar structure. Importantly, these simple lipid models are useful for investigating microstructure of the intercellular lipids in the stratum corneum.

Keyword: barrier function

Improving the physical and moisture properties of Lepidium perfoliatum seed gum biodegradable film with stearic and acids.

Stearic and fatty acids (10%, 20% and 30%, W/W gum) were used to improve the properties of Lepidium perfoliatum seed gum (LPSG) film. The impact of the incorporation of fatty acids into the film matrix was studied by investigating the physical, mechanical, and properties of the films. Addition of stearic and fatty acids to LPSG films reduced their water vapor permeability (WVP), moisture content, water solubility and water adsorption. Increasing fatty concentration from 10% to 30%, reduced the elongation at break (EB). Lower values of tensile strength (TS) and elastic modulus (EM) were obtained in the presence of higher fatty acids concentrations. Incorporation of fatty acids led to production of opaque films and the opacity increased as of fatty acids concentration. Results showed that moisture content, water solubility and WVP decreased as the chain length of fatty increased. Therefore, LPSG-fatty acids composite film could be used for packaging in which a low affinity toward water is needed.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: barrier function

From the Cover: Comparative Proteomics Reveals Silver Nanoparticles Alter Fatty Metabolism and Amyloid Beta Clearance for Neuronal Apoptosis in a Triple Cell Coculture Model of the Blood-Brain .

Silver nanoparticles (AgNPs) enter the central nervous system through the blood-brain (BBB). AgNP exposure can increase amyloid beta (Aβ) deposition in neuronal cells to potentially induce Alzheimer\'s disease (AD) progression. However, the mechanism through which AgNPs alter BBB permeability in endothelial cells and subsequently lead to AD progression remains unclear. This study investigated whether AgNPs disrupt the tight junction proteins of brain endothelial cells, and alter the proteomic metabolism of neuronal cells underlying AD progression in a triple cell coculture model constructed using mouse brain endothelial (bEnd.3) cells, mouse brain astrocytes (ALT), and mouse neuroblastoma neuro-2a (N2a) cells. The results showed that AgNPs accumulated in ALT and N2a cells because of the disruption of tight junction proteins, claudin-5 and ZO-1, in bEnd.3 cells. The proteomic profiling of N2a cells after AgNP exposure identified 298 differentially expressed proteins related to fatty metabolism. Particularly, AgNP-induced production was observed in N2a cells, which might promote Aβ generation. Moreover, AgNP exposure increased the protein expression of amyloid precursor protein (APP) and Aβ generation-related secretases, PSEN1, PSEN2, and β-site APP cleaving enzyme for APP cleavage in ALT and N2a cells, stimulated Aβ40 and Aβ42 secretion in the culture medium, and attenuated the gene expression of Aβ clearance-related receptors, P-gp and LRP-1, in bEnd.3 cells. Increased Aβ might further aggregate on the neuronal cell surface to enhance the secretion of inflammatory cytokines, MCP-1 and IL-6, thus inducing apoptosis in N2a cells. This study suggested that AgNP exposure might cause Aβ deposition and inflammation for subsequent neuronal cell apoptosis to potentially induce AD progression.© The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Keyword: barrier function

DHA upregulates FADS2 expression in primary cortical astrocytes exposed to vitamin A.

The fads2 gene encoding delta6-desaturase, the rate-limiting enzyme of the LCPUFA biosynthesis is expressed in astrocytes. Dietary fatty acids, which cross the blood-brain , may regulate the transcription of lipogenic enzymes through activation of transcription factors such as peroxisome proliferator-activated receptors (PPARs). The PPARs form the transcription complex with retinoid X receptors (RXRs) that are activated by 9-cis retinoic , a metabolite of vitamin A (VA). The study examines whether challenge of astrocytes with VA, prior 24-h treatment with (PA), alpha-linolenic (ALA) or docosahexaenoic (DHA) has the effect on the FADS2 expression. RT-qPCR showed that in astrocytes not challenged with VA, PA increased fads2 gene expression and DHA decreased it. However, in VA-primed astrocytes, PA doubled the FADS2 mRNA levels, while DHA increased fads2 gene expression, oppositely to non-primed cells. Furthermore, similar changes were seen in VA-primed astrocytes with regard to delta6-desaturase protein levels following PA and DHA treatment. ALA did not have any effect on the FADS2 mRNA and protein levels in either VA-primed or non-primed astrocytes. These findings indicate that in the presence of vitamin A, DHA upregulates fads2 gene expression in astrocytes.

Keyword: barrier function

Palmitoylethanolamide Regulates Production of Pro-Angiogenic Mediators in a Model of β Amyloid-Induced Astrogliosis In Vitro.

Aβ-induced astrogliosis can worsen the eziopathogenesis of Alzheimer disease (AD) by the release of proinflammatory and pro-oxidant mediators. Activated glial cells may release also pro-angiogenic molecules. The role of angiogenesis in AD is still controversial: although angiogenesis brings oxygen and nutrients to injured tissue, it may also exacerbate reactive gliosis. Moreover, by altering blood-brain permeability pro-angiogenic mediators promote passage of inflammatory/immune-competent cells into the brain, thereby exacerbating gliosis. The release of proangiogenic factors during astrogliosis may thus be a key-step in controlling AD progression. The endogenous fatty amide, palmitoylethanolamide (PEA), is a pleiotropic mediator exerting anti-inflammatory, antinociceptive and antiangiogenic effects in several in vitro and in vivo models of chronic-degenerative disease. In this study, we investigated the effects of PEA in AD angiogenesis and neuroinflammation by using conditioned medium from untreated and Aβ-treated C6 rat astroglioma cells and HUVEC human endothelial cells. PEA (10-8-10-6 M) concentration-dependently reduced expression of pro-inflammatory and pro-angiogenic markers in Aβ (1 μg/mL)-stimulated C6 cells. Moreover, culture medium from PEA-treated C6 cells reduced HUVEC cell proliferation as compared to cells treated with conditioned medium from Aβ-treated C6 cells. Immunocytochemical analysis revealed that PEA treatment inhibited nuclear levels of mitogen-activated protein kinase 1 (the main pro-angiogenic pathway) and cytoplasmic vascular endothelial growth factor in HUVEC cells receiving C6 conditioned medium. Finally, the peroxisome proliferator-activated receptor alpha inhibitor GW6471, added to Aβ-treated C6 cells blocked all PEA effects in this model, suggesting that PEA acts through a proliferator-activated receptor alpha-dependent mechanism on astroglial cells. Collectively, these data support the potential therapeutic utility of PEA in AD.

Keyword: barrier function

WAX INDUCER1 (HvWIN1) transcription factor regulates free fatty biosynthetic genes to reinforce cuticle to resist Fusarium head blight in barley spikelets.

Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most devastating diseases of wheat and barley. Resistance to FHB is highly complex and quantitative in nature, and is most often classified as resistance to spikelet infection and resistance to spread of pathogen through the rachis. In the present study, a resistant (CI9831) and a susceptible (H106-371) two-row barley genotypes, with contrasting levels of spikelet resistance to FHB, pathogen or mock-inoculated, were profiled for metabolites based on liquid chromatography and high resolution mass spectrometry. The key resistance-related (RR) metabolites belonging to fatty acids, phenylpropanoids, flavonoids and terpenoid biosynthetic pathways were identified. The free fatty acids (FFAs) linoleic and acids were among the highest fold change RR induced (RRI) metabolites. These FFAs are deposited as cutin monomers and oligomers to reinforce the cuticle, which acts as a to pathogen entry. Quantitative real-time PCR studies revealed higher expressions of KAS2, CYP86A2, CYP89A2, LACS2 and WAX INDUCER1 (HvWIN1) transcription factor in the pathogen-inoculated resistant genotype than in the susceptible genotype. Knockdown of HvWIN1 by virus-induced genes silencing (VIGS) in resistant genotype upon pathogen inoculation increased the disease severity and fungal biomass, and decreased the abundance of FFAs like linoleic and acids. Notably, the expression of CYP86A2, CYP89A2 and LAC2 genes was also suppressed, proving the link of HvWIN1 in regulating these genes in cuticle biosynthesis as a defense response.© The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Keyword: barrier function

Long-term exposition to a high fat diet favors the appearance of β-amyloid depositions in the brain of C57BL/6J mice. A potential model of sporadic Alzheimer\'s disease.

The sporadic and late-onset form of Alzheimer\'s disease (AD) constitutes the most common form of dementia. This non-familiar form could be a consequence of metabolic syndrome, characterized by obesity and the development of a brain-specific insulin resistance known as type III diabetes. This work demonstrates the development of a significant AD-like neuropathology due to these metabolic alterations.C57BL/6J mice strain were divided into two groups, one fed with a diet rich in (high-fat diet, HFD) since their weaning until 16 months of age, and another group used as a control with a regular diet. The analyses were carried out in the dentate gyrus area of the hippocampus using a Thioflavin-S stain and immunofluorescence assays.The most significant finding of the present research was that HFD induced the deposition of the βA peptide. Moreover, the diet also caused alterations in different cell processes, such as increased inflammatory reactions that lead to a decrease in the neuronal precursor cells. In addition, the results show that there were also dysregulations in normal autophagy and apoptosis, mechanisms related to βA formation.The present findings confirm that HFD favors the formation of βA depositions in the brain, a key feature of AD, supporting the metabolic hypothesis of sporadic AD.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: barrier function

Saturated and Unsaturated Fatty Acids Differently Modulate Colonic Goblet Cells In Vitro and in Rat Pups.

High-fat diets induce intestinal alterations and promote intestinal diseases. Little is known about the effects of long-chain fatty acids (LCFAs) on mucin 2 (MUC2) production by goblet cells, which are crucial for intestinal protection.We investigated the effects of LCFAs on the differentiation of colonic goblet cells, MUC2 expression, and colonic .Upon reaching confluence, human colonic mucus-secreting HT29-MTX cells were stimulated (21 d) with a saturated LCFA ( or stearic ), a monounsaturated LCFA (oleic ), or a polyunsaturated LCFA (linoleic, γ-linolenic, α-linolenic, or eicosapentaenoic ). In addition, rat pups underwent oral administration of oil (palm, rapeseed, or sunflower oil) or water (10 μL/g body weight, postnatal days 10-15). Subsequently, colon goblet cells were studied by Western blotting, reverse transcriptase-quantitative polymerase chain reaction, and immunohistochemistry and colonic transmucosal electrical resistance was measured by using Ussing chambers.In vitro, enhanced MUC2 production (140% of control) and hepatocyte nuclear factor 4α expression, whereas oleic, linoleic, γ-linolenic, α-linolenic, and eicosapentaenoic acids reduced MUC2 expression (at least -50% of control). All unsaturated LCFAs decreased the expression of human atonal homolog 1, a transcription factor controlling goblet cell differentiation (at least -31% vs. control). In vivo, rats fed palm oil had higher concentrations (3-fold) in their colonic contents and increased mucus granule surfaces in their goblet cells (>2-fold) than did all other groups. Palm oil also increased colonic transmucosal electrical resistance (245% of control), yet had no effect on occludin and zonula occludens-1 expression. In contrast, sunflower and rapeseed oils decreased goblet cell number when compared with control (at least -10%) and palm oil (at least -14%) groups.Palm oil in rat pups and in HT29-MTX cells increase the production of MUC2 and strengthen the intestinal . In contrast, unsaturated LCFAs decrease MUC2 expression. These data should be taken into account in the context of preventive or therapeutic nutritional programs.© 2015 American Society for Nutrition.

Keyword: barrier function

Kidney Proximal Tubule Lipoapoptosis Is Regulated by Fatty Transporter-2 (FATP2).

Albuminuria and tubular atrophy are among the highest risks for CKD progression to ESRD. A parsimonious mechanism involves leakage of albumin-bound nonesterified fatty acids (NEFAs) across the damaged glomerular filtration and subsequent reabsorption by the downstream proximal tubule, causing lipoapoptosis. We sought to identify the apical proximal tubule transporter that mediates NEFA uptake and cytotoxicity. We observed transporter-mediated uptake of fluorescently labeled NEFA in cultured proximal tubule cells and microperfused rat proximal tubules, with greater uptake from the apical surface than from the basolateral surface. Protein and mRNA expression analyses revealed that kidney proximal tubules express transmembrane fatty transporter-2 (FATP2), encoded by , but not the other candidate transporters CD36 and free fatty receptor 1. Kidney FATP2 localized exclusively to proximal tubule epithelial cells along the apical but not the basolateral membrane. Treatment of mice with lipidated albumin to induce proteinuria caused a decrease in the proportion of tubular epithelial cells and an increase in the proportion of interstitial space in kidneys from wild-type but not mice. microperfusion and experiments with NEFA-bound albumin at concentrations that mimic apical proximal tubule exposure during glomerular injury revealed significantly reduced NEFA uptake and palmitate-induced apoptosis in microperfused proximal tubules and or FATP2 shRNA-treated proximal tubule cell lines compared with wild-type or scrambled oligonucleotide-treated cells, respectively. We conclude that FATP2 is a major apical proximal tubule NEFA transporter that regulates lipoapoptosis and may be an amenable target for the prevention of CKD progression.Copyright © 2018 by the American Society of Nephrology.

Keyword: barrier function

Sexually dimorphic brain fatty composition in low and high fat diet-fed mice.

In this study, we analyzed the fatty profile of brains and plasma from male and female mice fed chow or a western-style high fat diet (WD) for 16 weeks to determine if males and females process fatty acids differently. Based on the differences in fatty acids observed in\xa0vivo, we performed in\xa0vitro experiments on N43 hypothalamic neuronal cells to begin to elucidate how the fatty milieu may impact brain inflammation.Using a comprehensive mass spectrometry fatty analysis, which includes a profile for 52 different fatty isomers, we assayed the plasma and brain fatty composition of age-matched male and female mice maintained on chow or a WD. Additionally, using the same techniques, we determined the fatty composition of N43 hypothalamic cells following exposure to and linoleic , alone or in combination.Our data demonstrate there is a sexual dimorphism in brain fatty content both following the consumption of the chow diet, as well as the WD, with males having an increased percentage of saturated fatty acids and reductions in ω6-polyunsaturated fatty acids when compared to females. Interestingly, we did not observe a sexual dimorphism in fatty content in the plasma of the same mice. Furthermore, exposure of N43 cells to the ω6-PUFA linoleic , which is higher in female brains when compared to males, reduces -induced inflammation.Our data suggest male and female brains, and not plasma, differ in their fatty profile. This is the first time, to our knowledge, lipidomic analyses has been used to directly test the hypothesis there is a sexual dimorphism in brain and plasma fatty composition following consumption of the chow diet, as well as following exposure to the WD.

Keyword: barrier function

AIF loss deregulates hematopoiesis and reveals different adaptive metabolic responses in bone marrow cells and thymocytes.

Mitochondrial metabolism is a tightly regulated process that plays a central role throughout the lifespan of hematopoietic cells. Herein, we analyze the consequences of the mitochondrial oxidative phosphorylation (OXPHOS)/metabolism disorder associated with the cell-specific hematopoietic ablation of apoptosis-inducing factor (AIF). AIF-null (AIF ) mice developed pancytopenia that was associated with hypocellular bone marrow (BM) and thymus atrophy. Although myeloid cells were relatively spared, the B-cell and erythroid lineages were altered with increased frequencies of precursor B cells, pro-erythroblasts I, and basophilic erythroblasts II. T-cell populations were dramatically reduced with a thymopoiesis blockade at a double negative (DN) immature state, with DN1 accumulation and delayed DN2/DN3 and DN3/DN4 transitions. In BM cells, the OXPHOS/metabolism dysfunction provoked by the loss of AIF was counterbalanced by the augmentation of the mitochondrial biogenesis and a shift towards anaerobic glycolysis. Nevertheless, in a caspase-independent process, the resulting excess of reactive oxygen species compromised the viability of the hematopoietic stem cells (HSC) and progenitors. This led to the progressive exhaustion of the HSC pool, a reduced capacity of the BM progenitors to differentiate into colonies in methylcellulose assays, and the absence of cell-autonomous HSC repopulating potential in vivo. In contrast to BM cells, AIF thymocytes compensated for the OXPHOS breakdown by enhancing fatty β-oxidation. By over-expressing CPT1, ACADL and PDK4, three key enzymes facilitating fatty β-oxidation (e.g., assimilation), the AIF thymocytes retrieved the ATP levels of the AIF cells. As a consequence, it was possible to significantly reestablish AIF thymopoiesis in vivo by feeding the animals with a high-fat diet complemented with an antioxidant. Overall, our data reveal that the mitochondrial signals regulated by AIF are critical to hematopoietic decision-making. Emerging as a link between mitochondrial metabolism and hematopoietic cell fate, AIF-mediated OXPHOS regulation represents a target for the development of new immunomodulatory therapeutics.

Keyword: barrier function

Renal Lipotoxicity-Associated Inflammation and Insulin Resistance Affects Actin Cytoskeleton Organization in Podocytes.

In the last few decades a change in lifestyle has led to an alarming increase in the prevalence of obesity and obesity-associated complications. Obese patients are at increased risk of developing hypertension, heart disease, insulin resistance (IR), dyslipidemia, type 2 diabetes and renal disease. The excess calories are stored as triglycerides in adipose tissue, but also may accumulate ectopically in other organs, including the kidney, which contributes to the damage through a toxic process named lipotoxicity. Recently, the evidence suggests that renal lipid accumulation leads to glomerular damage and, more specifically, produces dysfunction in podocytes, key cells that compose and maintain the glomerular filtration . Our aim was to analyze the early mechanisms underlying the development of renal disease associated with the process of lipotoxicity in podocytes. Our results show that treatment of podocytes with produced intracellular accumulation of lipid droplets and abnormal glucose and lipid metabolism. This was accompanied by the development of inflammation, oxidative stress and endoplasmic reticulum stress and insulin resistance. We found specific rearrangements of the actin cytoskeleton and slit diaphragm proteins (Nephrin, P-Cadherin, Vimentin) associated with this insulin resistance in -treated podocytes. We conclude that lipotoxicity accelerates glomerular disease through lipid accumulation and inflammation. Moreover, saturated fatty acids specifically promote insulin resistance by disturbing the cytoarchitecture of podocytes. These data suggest that renal lipid metabolism and cytoskeleton rearrangements may serve as a target for specific therapies aimed at slowing the progression of podocyte failure during metabolic syndrome.

Keyword: barrier function

Naturally occurring ω-Hydroxyacids.

ω-Hydroxyacids are fatty acids bearing a hydroxyl group on the terminal carbon. They are found in mammals and higher plants and are often involved in providing a permeability , the primary purpose of which is to reduce water loss. Some ω-hydroxyacid derivatives may be involved in waterproofing and signalling. The purpose of this review was to survey the known natural sources of ω-hydroxyacids. ω-Hydroxyacids are produced by two different P450-dependent mechanisms. The longer (30-34 carbons) ω-hydroxyacids are produced by chain extension from until the chain extends across the membrane in which the extension is taking place, and then the terminal carbon is hydroxylated. Shorter fatty acids can be hydroxylated directly to produce C16 and C18 ω-hydroxyacids found in plants and 20-eicosatetraenoic (20-HETE) by a different P450. The C16 and C18 ω-hydroxyacids are components of polymers in plants. The long-chain ω-hydroxyacids are found in epidermal sphingolipids, in giant-ring lactones from the sebum of members of the equidae, as a component of meibum and in carnauba wax and wool wax.© 2017 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Keyword: barrier function

Metabolomic profiles reveal key metabolic changes in heat stress-treated mouse Sertoli cells.

Heat stress (HS) is a potential harmful factor for male reproduction. However, the effect of HS on Sertoli cells is largely unknown. In this study, the metabolic changes in Sertoli cell line were analyzed after HS treatment. Metabolomic analysis revealed that carnitine, 2-hydroxy , nicotinic , niacinamide, adenosine monophosphate, glutamine and creatine were the key changed metabolites. We found the expression levels of BTB factors (Connexin43, ZO-1, Vimentin, Claudin1, Claudin5) were disrupted in TM-4 cells after HS treatment, which were recovered by the addition of carnitine. RT-PCR indicated that the mRNA levels of inflammatory cytokines (IL-1α, IL-1β, IL-6) were increased after HS treatment, and their related miRNAs (miR-132, miR-431, miR-543) levels were decreased. Our metabolomic data provided a novel understanding of metabolic changes in male reproductive cells after HS treatment and revealed that HS-induced changes of BTB factors and inflammatory status might be caused by the decreased carnitine after HS treatment.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: barrier function

Sebum lipids influence macrophage polarization and activation.

As lipids are known to regulate macrophage functions, it is reasonable to suppose that a sebocyte-macrophage axis mediated by sebum lipids may exist.To investigate if sebocytes could contribute to the differentiation, polarization and of macrophages with their secreted lipids.Oil Red O lipid staining and Raman spectroscopy were used to assess the dermal lipid content and penetration. Immunohistochemistry was used to analyse the macrophage subsets. Human peripheral blood monocytes were differentiated in the presence of either supernatant from human SZ95 sebocytes or major sebum lipid components and activated with Propionibacterium acnes. Macrophage surface markers and their capacity to uptake fluorescein isothiocyanate-conjugated P.\xa0acnes were detected by fluorescence-activated cell sorting measurements. Cytokine protein levels were evaluated by enzyme-linked immunosorbent assay and Western blot analysis.Sebaceous gland-rich skin had an increased dermal lipid content vs. sebaceous gland-poor skin to which all the tested sebum component lipids could contribute by penetrating the dermoepidermal . Of the lipids, oleic and linoleic promoted monocyte differentiation into alternatively activated macrophages. Moreover, linoleic also had an anti-inflammatory effect in P.\xa0acnes-activated macrophages, inhibiting the secretion of interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α. Squalene, , stearic and oleic augmented the secretion of IL-1β, even in the absence of P. acnes, whereas oleic had a selective effect of inducing IL-1β but downregulating IL-6 and TNF-α secretion.Our results suggest a role for sebaceous glands in modulating innate immune responses via their secreted lipids that are of possible pathological and therapeutic relevance.© 2017 British Association of Dermatologists.

Keyword: barrier function

Effects of elevated growth temperature and heat shock on the lipid composition of the inner and outer membranes of Yersinia pseudotuberculosis.

Differences in the distribution of individual phospholipids between the inner (IM) and outer membranes (OM) of gram-negative bacteria have been detected in mesophilic Escherichia, Erwinia and Salmonella species but have never been investigated in the psychrotrophic Yersinia genus. Therefore, the influence of an elevated growth temperature and heat shock on the phospholipid and fatty (FA) compositions of the fractionated Yersinia pseudotuberculosis envelope was investigated. The shift of the growth temperature from 8\xa0°C to 37\xa0°C to mimic the switch from saprophytic to parasitic growth of this bacteria and the exposure of the cells to heat shock, which was induced by a sharp increase in the temperature from 8\xa0°C to 45\xa0°C, increased the lysophosphatidylethanolamine content from zero and 1% to 6% and 10% in the IM and OM, respectively. These changes were accompanied by a decrease in the phosphatidylethanolamine (PE) content and a drastic increase (up to 3-fold higher) in the phosphatidylglycerol (PG) level in the OM of the bacteria, which increases the net negative charge of the cell envelope. The levels of the predominant saturated (16:0) and cyclopropane FAs were approximately 1.5- and 7.5-fold higher, respectively, but the content of the predominant unsaturated palmitoleic (16:1n-7) and cis-vaccenic (18:1n-7) FAs was approximately 10-30-fold lower in both membranes that were isolated from the cells grown at elevated temperatures. Due to these changes, reflecting the process of "homeoviscous adaptation", the ratio between the unsaturated and saturated FAs decreased but remained higher in the IM than that in the OM. Simultaneously, no significant changes were observed in the FA composition of cells subjected to heat shock, demonstrating a difference between the responses of the heat-shocked and heat-adapted Y.\xa0pseudotuberculosis. The unique ability of Y.\xa0pseudotuberculosis to reciprocally regulate the ratio of anionic PG and net neutral PE and therefore adjust the negative charge of the OM may be a common strategy used by pathogenic bacteria to promote the of the OM.Copyright © 2016. Published by Elsevier B.V.

Keyword: barrier function

TNF-α stimulates endothelial transcytosis and promotes insulin resistance.

Persistent elevation of plasma TNF-α is a marker of low grade systemic inflammation. (PA) is the most abundant type of saturated fatty in human body. PA is bound with albumin in plasma and could not pass through endothelial freely. Albumin-bound PA has to be transported across monolayer endothelial cells through intracellular transcytosis, but not intercellular diffusion. In the present study, we discovered that TNF-α might stimulate PA transcytosis across cardiac microvascular endothelial cells, which further impaired the insulin-stimulated glucose uptake by cardiomyocytes and promoted insulin resistance. In this process, TNF-α-stimulated endothelial autophagy and NF-κB signaling crosstalk with each other and orchestrate the whole event, ultimately result in increased expression of fatty transporter protein 4 (FATP4) in endothelial cells and mediate the increased PA transcytosis across microvascular endothelial cells. Hopefully the present study discovered a novel missing link between low grade systemic inflammation and insulin resistance.

Keyword: barrier function

2-Chlorohexadecanoic induces ER stress and mitochondrial dysfunction in brain microvascular endothelial cells.

Peripheral leukocytes induce blood-brain (BBB) dysfunction through the release of cytotoxic mediators. These include hypochlorous (HOCl) that is formed via the myeloperoxidase-HO-chloride system of activated phagocytes. HOCl targets the endogenous pool of ether phospholipids (plasmalogens) generating chlorinated inflammatory mediators like e.g. 2-chlorohexadecanal and its conversion product 2-chlorohexadecanoic (2-ClHA). In the cerebrovasculature these compounds inflict damage to brain microvascular endothelial cells (BMVEC) that form the morphological basis of the BBB. To follow subcellular trafficking of 2-ClHA we synthesized a \'clickable\' alkyne derivative (2-ClHyA) that phenocopied the biological activity of the parent compound. Confocal and superresolution structured illumination microscopy revealed accumulation of 2-ClHyA in the endoplasmic reticulum (ER) and mitochondria of human BMVEC (hCMEC/D3 cell line). 2-ClHA and its alkyne analogue interfered with protein palmitoylation, induced ER-stress markers, reduced the ER ATP content, and activated transcription and secretion of interleukin (IL)-6 as well as IL-8. 2-ClHA disrupted the mitochondrial membrane potential and induced procaspase-3 and PARP cleavage. The protein kinase R-like ER kinase (PERK) inhibitor GSK2606414 suppressed 2-ClHA-mediated activating transcription factor 4 synthesis and IL-6/8 secretion, but showed no effect on endothelial dysfunction and cleavage of procaspase-3. Our data indicate that 2-ClHA induces potent lipotoxic responses in brain endothelial cells and could have implications in inflammation-induced BBB dysfunction.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: barrier function

Position of Proline Mediates the Reactivity of S-Palmitoylation.

Palmitoylation, a post-translational modification in which a saturated 16-carbon chain is added predominantly to a cysteine residue, participates in various biological functions. The position of proline relative to other residues being post-translationally modified has been previously reported as being important. We determined that proline is statistically enriched around cysteines known to be S-palmitoylated. The goal of this work was to determine how the position of proline influences the palmitoylation of the cysteine residue. We established a mass spectrometry-based approach to investigate time- and temperature-dependent kinetics of autopalmitoylation in vitro and to derive the thermodynamic parameters of the transition state associated with palmitoylation; to the best of our knowledge, our work is the first to study the kinetics and activation properties of the palmitoylation process. We then used these thermochemical parameters to determine if the position of proline relative to the modified cysteine is important for palmitoylation. Our results show that peptides with proline at the -1 position of cysteine in their sequence (PC) have lower enthalpic barriers and higher entropic barriers in comparison to the same peptides with proline at the +1 position of cysteine (CP); interestingly, the free-energy barriers for both pairs are almost identical. Molecular dynamics studies demonstrate that the flexibility of the cysteine backbone in the PC-containing peptide when compared to the CP-containing peptide explains the increased entropic and decreased enthalpic observed experimentally.

Keyword: barrier function

Profiling of epidermal lipids in a mouse model of dermatitis: Identification of potential biomarkers.

Lipids are important structural and functional components of the skin. Alterations in the lipid composition of the epidermis are associated with inflammation and can affect the of the skin. SHARPIN-deficient cpdm mice develop a chronic dermatitis with similarities to atopic dermatitis in humans. Here, we used a recently-developed approach named multiple reaction monitoring (MRM)-profiling and single ion monitoring to rapidly identify discriminative lipid ions. Shorter fatty acyl residues and increased relative amounts of sphingosine ceramides were observed in cpdm epidermis compared to wild type mice. These changes were accompanied by downregulation of the Fasn gene which encodes fatty synthase. A profile of diverse lipids was generated by fast screening of over 300 transitions (ion pairs). Tentative attribution of the most significant transitions was confirmed by product ion scan (MS/MS), and the MRM-profiling linear intensity response was validated with a C17-ceramide lipid standard. Relative quantification of sphingosine ceramides CerAS(d18:1/24:0)2OH, CerAS(d18:1/16:0)2OH and CerNS(d18:1/16:0) discriminated between the two groups with 100% accuracy, while the free fatty acids cerotic , 16-hydroxy , and docosahexaenoic (DHA) had 96.4% of accuracy. Validation by liquid chromatography tandem mass spectrometry (LC-MS/MS) of the above-mentioned ceramides was in agreement with MRM-profiling results. Identification and rapid monitoring of these lipids represent a tool to assess therapeutic outcomes in SHARPIN-deficient mice and other mouse models of dermatitis and may have diagnostic utility in atopic dermatitis.

Keyword: barrier function

Autophagy Protects against -Induced Apoptosis in Podocytes in vitro.

Autophagy is a highly conserved degradation process that is involved in the clearance of proteins and damaged organelles to maintain intracellular homeostasis and cell integrity. Type 2 diabetes is often accompanied by dyslipidemia with elevated levels of free fatty acids (FFAs). Podocytes, as an important component of the filtration , are susceptible to lipid disorders. The loss of podocytes causes proteinuria, which is involved in the pathogenesis of diabetic nephropathy. In the present study, we demonstrated that (PA) promoted autophagy in podocytes. We further found that PA increased the production of reactive oxygen species (ROS) in podocytes and that NAC (N-acetyl-cysteine), a potent antioxidant, significantly eliminated the excessive ROS and suppressed autophagy, indicating that the increased generation of ROS was associated with the -induced autophagy in podocytes. Moreover, we also found that PA stimulation decreased the mitochondrial membrane potential in podocytes and induced podocyte apoptosis, while the inhibition of autophagy by chloroquine (CQ) enhanced -induced apoptosis accompanied by increased ROS generation, and the stimulation of autophagy by rapamycin (Rap) remarkably suppressed -induced ROS generation and apoptosis. Taken together, these in vitro findings suggest that PA-induced autophagy in podocytes is mediated by ROS production and that autophagy plays a protective role against PA-induced podocyte apoptosis.

Keyword: barrier function

Dynamic alterations in the gut microbiota and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut microbiota and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal was more impaired. Compared with the control diet, the MCD diet induced gradual gut microbiota dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut microbiota and metabolome.

Keyword: barrier function

Modulation of stratum corneum lipid composition and organization of human skin equivalents by specific medium supplements.

Our in-house human skin equivalents contain all stratum corneum (SC) lipid classes, but have a reduced level of free fatty acids (FAs), of which a part is mono-unsaturated. These differences lead to an altered SC lipid organization and thereby a reduced compared to human skin. In this study, we aimed to improve the SC FA composition and, consequently, the SC lipid organization of the Leiden epidermal model (LEM) by specific medium supplements. The standard FA mixture (consisting of , linoleic and arachidonic acids) supplemented to the medium was modified, by replacing protonated with deuterated or by the addition of deuterated arachidic to the mixture, to determine whether FAs are taken up from the medium and are incorporated into SC of LEM. Furthermore, supplementation of the total FA mixture or that of alone was increased four times to examine whether this improves the SC FA composition and lipid organization of LEM. The results demonstrate that the deuterated FAs are taken up into LEMs and are subsequently elongated and incorporated in their SC. However, a fourfold increase in supplementation does not change the SC FA composition or lipid organization of LEM. Increasing the concentration of the total FA mixture in the medium resulted in a decreased level of very long chain FAs and an increased level of mono-unsaturated FAs, which lead to deteriorated SC lipid properties. These results indicate that SC lipid properties can be modulated by specific medium supplements.© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: barrier function

Fatty acids penetration into human skin ex vivo: A TOF-SIMS analysis approach.

Linoleic, oleic, palmitoleic, , and stearic fatty acids (FAs) are commonly used in dermatological formulations. They differ by their structure, presence in the skin, and mode of application in pharmaceuticals and cosmetics compounding. These FAs are also known as chemical penetration enhancers, but their mechanisms of penetration enhancement and effect on characteristics of the skin require additional study. In this study, the authors conducted an ex vivo analysis of the distribution of lipid components in the epidermis and dermis of human skin after applying individual FAs. The goal was to elucidate possible mechanisms of penetration enhancement and FA effects on characteristics of the skin. FA penetration studies were conducted ex vivo on human skin and time-of-flight secondary ion mass spectrometry (TOF-SIMS) bioimaging analysis was performed to visualize and analyze distribution of FAs in skin sections. The current study demonstrated that TOF-SIMS imaging was effective in visualizing the distribution of linoleic, oleic, palmitoleic, , and stearic in the human skin ex vivo after the skin penetration experiment of individual FAs. The integration of the obtained TOF-SIMS images allowed a semiquantitative comparison of the effects induced by individual FA applications on the human skin ex vivo. FAs showed varying abilities to penetrate the skin and disorder the FAs within the skin, based on their structures and physicochemical properties. Linoleic penetrated the skin and changed the distribution of all the analyzed FAs. Skin treatment with palmitoleic or oleic increased the amounts of singular FAs in the skin. Penetration of saturated FAs was low, but it increased the detected amounts of linoleic in both skin layers. The results indicate that application of FAs on the skin surface induce redistribution of native FAs not only in the stratum corneum layer of epidermis but also in the lipid content of full epidermis and dermis layers. The results indicate that topically applied pharmaceutical products should be evaluated for potential chemical penetration enhancement and lipid component redistribution effects during formulation.

Keyword: barrier function

2-Chlorofatty acids induce Weibel-Palade body mobilization.

Endothelial dysfunction is a hallmark of multiple inflammatory diseases. Leukocyte interactions with the endothelium have significant effects on vascular wall biology and pathophysiology. Myeloperoxidase (MPO)-derived oxidant products released from leukocytes are potential mediators of inflammation and endothelial dysfunction. 2-Chlorofatty acids (2-ClFAs) are produced as a result of MPO-derived HOCl targeting plasmalogen phospholipids. Chlorinated lipids have been shown to be associated with multiple inflammatory diseases, but their impact on surrounding endothelial cells has not been examined. This study tested the biological properties of the 2-ClFA molecular species 2-chlorohexadecanoic (2-ClHA) on endothelial cells. A synthetic alkyne analog of 2-ClHA, 2-chlorohexadec-15-ynoic (2-ClHyA), was used to examine the subcellular localization of 2-ClFA in human coronary artery endothelial cells. Click chemistry experiments revealed that 2-ClHyA localizes to Weibel-Palade bodies. 2-ClHA and 2-ClHyA promote the release of P-selectin, von Willebrand factor, and angiopoietin-2 from endothelial cells. Functionally, 2-ClHA and 2-ClHyA cause neutrophils to adhere to and platelets to aggregate on the endothelium, as well as increase permeability of the endothelial which has been tied to the release of angiopoietin-2. These findings suggest that 2-ClFAs promote endothelial cell dysfunction, which may lead to broad implications in inflammation, thrombosis, and blood vessel stability.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: barrier function

Lipid-based nanocarrier efficiently delivers highly water soluble drug across the blood-brain into brain.

Delivering highly water soluble drugs across blood-brain (BBB) is a crucial challenge for the formulation scientists. A successful therapeutic intervention by developing a suitable drug delivery system may revolutionize treatment across BBB. Efforts were given here to unravel the capability of a newly developed fatty combination (stearic :oleic :\u2009=\u20098.08:4.13:1) (ML) as fundamental component of nanocarrier to deliver highly water soluble zidovudine (AZT) as a model drug into brain across BBB. A comparison was made with an experimentally developed standard phospholipid-based nanocarrier containing AZT. Both the formulations had nanosize spherical unilamellar vesicular structure with highly negative zeta potential along with sustained drug release profiles. Gamma scintigraphic images showed both the radiolabeled formulations successfully crossed BBB, but longer retention in brain was observed for ML-based formulation (MGF) as compared to soya lecithin (SL)-based drug carrier (SYF). Plasma and brain pharmacokinetic data showed less clearance, prolonged residence time, more bioavailability and sustained release of AZT from MGF in rats compared to those data of the rats treated with SYF/AZT suspension. Thus, ML may be utilized to successfully develop drug nanocarrier to deliver drug into brain across BBB, in a sustained manner for a prolong period of time and may provide an effective therapeutic strategy for many diseases of brain. Further, many anti-HIV drugs cannot cross BBB sufficiently. Hence, the developed formulation may be a suitable option to carry those drugs into brain for better therapeutic management of HIV.

Keyword: barrier function

Docosahexaenoic and Eicosapentaenoic Acids Prevent Altered-Muc2 Secretion Induced by by Alleviating Endoplasmic Reticulum Stress in LS174T Goblet Cells.

Diets high in saturated fatty acids (FA) represent a risk factor for the development of obesity and associated metabolic disorders, partly through their impact on the epithelial cell . We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus . To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or -9, -6, or -3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic nor -9 and -6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 diabetes and obesity or inflammatory bowel disease. These in vitro data remain to be explored in vivo in a context of dietary obesity.

Keyword: barrier intergrity

Fatty ethyl esters induce intestinal epithelial dysfunction via a reactive oxygen species-dependent mechanism in a three-dimensional cell culture model.

Evidence is accumulating that ethanol and its oxidative metabolite, acetaldehyde, can disrupt intestinal epithelial integrity, an important factor contributing to ethanol-induced liver injury. However, ethanol can also be metabolized non-oxidatively generating phosphatidylethanol and fatty ethyl esters (FAEEs). This study aims to investigate the effects of FAEEs on , and to explore the role of oxidative stress as possible mechanism.Epithelial permeability was assessed by paracellular flux of fluorescein isothiocyanate-conjugated dextran using live cell imaging. Cell integrity was evaluated by lactate dehydrogenase release. Localization and protein levels of ZO-1 and occludin were analyzed by immunofluorescence and cell-based ELISA, respectively. Intracellular oxidative stress and cellular ATP levels were measured by dichlorofluorescein and luciferase driven bioluminescence, respectively.In vitro, ethyl oleate and ethyl palmitate dose dependently increased permeability associated with disruption and decreased ZO-1 and occludin protein levels, respectively, and increased intracellular oxidative stress without compromising cell viability. These effects could partially be attenuated by pretreatment with the antioxidant, resveratrol, pointing to the role of oxidative stress in the FAEEs-induced intestinal dysfunction.These findings show that FAEEs can induce intestinal dysfunction by disrupting the tight junctions, most likely via reactive oxygen species-dependent mechanism.

Keyword: barrier intergrity

Selective disruption of endothelial function in culture by pure fatty acids and fatty acids derived from animal and plant fats.

Endothelial cell has been suggested to play a role in the development of atherosclerosis. The effects of fatty acids on endothelial function were tested by measuring albumin transport across endothelial monolayers cultured on polycarbonate filters. Compared with control cultures, a 24-h exposure to 90 mumol/L lauric (12:0) and linoleic (18:2) but not to butyric (4:0), hexanoic (6:0), octanoic (8:0), decanoic (10:0), myristic (14:0), (16:0) or stearic (18:0) caused an increase in albumin transfer across endothelial monolayers. Selective enrichment of a "physiological" serum fatty mixture (FA-Mix; 90 mumol/L) with 90 mumol/L of 12:0 or 18:2 significantly increased albumin transfer, whereas enrichment with 90 mumol/L of 4:0, 16:0 or 18:0 significantly decreased albumin transfer relative to 180 mumol/L FA-Mix. Only 12:0- or 18:2-treated cultures showed increased Ca(++)-ATPase activity and the presence of lipid droplets. Fatty acids (60 mumol/L) extracted from butter fat and beef tallow had no effect on albumin transfer, whereas fatty acids extracted from chicken fat and corn oil consistently disrupted endothelial function. This fat-induced disruption of endothelial function seems to be related to the amount of 18:2 present in each fat source. These data indicate that unsaturated fats cause cellular perturbations that result in a decrease in endothelial function in this model system, and that high dietary levels of unsaturated fats may be detrimental to cell .

Keyword: barrier intergrity

Single Triglyceride-Rich Meal Destabilizes Functions and Initiates Inflammatory Processes of Endothelial Cells.

Postprandial hypertriglyceridemia is an independent risk factor for cardiovascular disease. The aim of this study was to assess the effects of a single fat-rich meal on functions and inflammatory status on human umbilical vascular endothelial cells (HUVECs), furthermore we assess the effects of mixture of and 25-hydroxycholesterol (PA +25OHCH) on of endothelial cells and their inflammatory properties. HUVECs were induced with serum of healthy volunteers taken before, and 3\u2009h after, the consumption of a meal with a standardized daily required dose of fats. In addition, endothelial cells were induced with PA +25OHCH (800\u2009μM/L+10\u2009μg/mL). Total cholesterol, triglycerides (TGs), high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, high sensitivity c-reactive protein, and glucose were measured at fasting and postprandially. HUVEC was measured in the RTCA-DP xCELLigence system. mRNA expression of interleukin (IL)-33, IL-32, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1), CX3C-chemokine, vascular endothelial growth factor (VEGF) occludin, and VE-cadherin was analyzed by real-time polymerase chain reaction. Viability and apoptosis were assessed in flow cytometry. The level of VEGF and IL-33 in fasting and postprandial serum was assessed by enzyme-linked immunosorbent assay (ELISA). Three hours after consumption of a fatty meal, all patients displayed increased levels of TGs and Toll-like receptors (110\u2009±\u200937\u2009mg/dL versus 182\u2009±\u200964\u2009mg/dL \u2009<\u20090.05) (24\u2009±\u200911\u2009mg/dL versus 42\u2009±\u200914\u2009mg/dL \u2009<\u20090.05). Postprandial serum and PA +25OHCH caused >20% decrease of HUVEC than fasting serum (\u2009<\u20090.001). HUVEC disintegration was accompanied by a decrease of occludin mRNA expression as compared with fasting serum (\u2009<\u20090.05). The fatty meal affected neither VE-cadherin mRNA expression nor its apoptosis (\u2009>\u20090.05). Mixture of PA +25OHCH caused decrease of VE-cadherin mRNA expression as compared with fasting serum (\u2009<\u20090.01). PA +25OHCH did not affect HUVEC apoptosis (\u2009>\u20090.05). Postprandial serum and PA +25OHCH caused increase of IL-33, MCP-1, ICAM-1, IL-32, VEGF, and CX3C-chemokine mRNA expression as compared with fasting serum (\u2009<\u20090.05). Moreover, level of VEGF in fatty serum was significantly higher (\u2009<\u20090.001). Postprandial lipemia after a single fatty meal may destabilize the endothelial and initiate inflammatory processes.

Keyword: barrier intergrity

Generation of free fatty acids from phospholipids regulates stratum corneum acidification and .

There is evidence that the " mantle" of the stratum corneum is important for both permeability formation and cutaneous antimicrobial defense. The origin of the acidic pH of the stratum corneum remains conjectural, however. Both passive (e.g., eccrine/sebaceous secretions, proteolytic) and active (e.g., proton pumps) mechanisms have been proposed. We assessed here whether the free fatty pool, which is derived from phospholipase-mediated hydrolysis of phospholipids during cornification, contributes to stratum corneum acidification and function. Topical applications of two chemically unrelated secretory phospholipase sPLA2 inhibitors, bromphenacylbromide and 1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol, for 3 d produced an increase in the pH of murine skin surface that was paralleled not only by a permeability abnormality but also altered stratum corneum (number of strippings required to break the ) and decreased stratum corneum cohesion (protein weight removed per stripping). Not only stratum corneum pH but also all of the functional abnormalities normalized when either , stearic, or linoleic acids were coapplied with the inhibitors. Moreover, exposure of intact murine stratum corneum to a neutral pH for as little as 3 h produced comparable abnormalities in stratum corneum and cohesion, and further amplified the inhibitor-induced functional alterations. Furthermore, short-term applications of an acidic pH buffer to inhibitor-treated skin also reversed the abnormalities in stratum corneum and cohesion, despite the ongoing decrease in free fatty levels. Finally, the secretory-phospholipase-inhibitor-induced alterations in /cohesion were in accordance with premature dissolution of desmosomes, demonstrated both by electron microscopy and by reduced desmoglein 1 levels in the stratum corneum (shown by immunofluorescence staining and visualized by confocal microscopy). Together, these results demonstrate: (i) the importance of phospholipid-to-free-fatty- processing for normal stratum corneum acidification; and (ii) the potentially important role of this pathway not only for homeostasis but also for the dual functions of stratum corneum and cohesion.

Keyword: barrier intergrity

Palmitoylation of brain capillary proteins.

Palmitoylation is a reversible posttranslational modification which is involved in the regulation of several membrane proteins such as beta 2-adrenergic receptor, p21ras and trimeric G-protein alpha-subunits. This covalent modification could be involved in the regulation of the numerous membrane proteins present in the blood-brain capillaries. The palmitoylation activity present in brain capillaries was characterized using [3H]palmitate labeling followed by chloroform methanol precipitation. Palmitate solubilizing agents such as detergents and bovine serum albumin (BSA), were used for optimizing activity. Some palmitoylated substrates were identified using [3H]palmitate labeling followed by immunoprecipitation with specific antibodies. Two optimal palmitate solubilization conditions were found, one involves cell permeabilization (Triton X-100) and the other represents a more physiological condition where membrane is conserved (BSA). Sensitivity to the cysteine modifier N-ethylmaleimide and to hydrolysis, using hydroxylamine or alkaline methanolysis, indicated that was bound to the proteins by a thioester bond. Maximal palmitate incorporation was reached after 30 or 60 min of incubation in the presence of Triton or BSA, respectively. Depalmitoylation was observed in the presence of BSA, but not with detergents. The palmitoylation reaction was optimal at pH 8 or 9 in the presence of Triton or BSA, respectively, but palmitoylated substrates were detectable over a wide range of pH values. In the presence of Triton X-100, the addition of ATP, CoA and Mg2+ to the incubation medium increased palmitoylation by up to 80-fold. Two palmitoylated substrates were identified, a 42 kDa G-protein alpha subunit and p21ras. The study shows that the utilization of palmitate solubilizing agents is essential to measure in vitro palmitoylation in brain capillaries. Several palmitoylated proteins are present in the blood-brain including five major substrates of 12, 21, 35, 42 and 55 kDa. It is suggested that palmitoylation could play a crucial role in the regulation of brain capillary function, since the two substrates identified in this study are known to be involved in signal transduction, vesicular transport and cell differentiation.

Keyword: barrier intergrity

damages gut epithelium and initiates inflammatory cytokine production.

The mechanisms leading to the low-grade inflammation observed during obesity are not fully understood. Seeking the initiating events, we tested the hypothesis that the intestine could be damaged by repeated lipid supply and therefore participate in inflammation. In mice, 1-5 palm oil gavages increased intestinal permeability via decreased expression and mislocalization of junctional proteins at the cell-cell contacts; altered the intestinal bacterial species by decreasing the abundance of Akkermansia muciniphila, segmented filamentous bacteria, and Clostridium leptum; and increased inflammatory cytokine expression. This was further studied in human intestinal epithelial Caco-2/TC7 cells using the two main components of palm oil, i.e., and oleic . Saturated impaired paracellular permeability and junctional protein localization, and induced inflammatory cytokine expression in the cells, but unsaturated oleic did not. Inhibiting de novo ceramide synthesis prevented part of these effects. Altogether, our data show that short exposure to palm oil or induces intestinal dysfunctions targeting and inflammation. Excessive palm oil consumption could be an early player in the gut alterations observed in metabolic diseases.Copyright © 2019. Published by Elsevier B.V.

Keyword: barrier intergrity

Autophagy Protects against -Induced Apoptosis in Podocytes in vitro.

Autophagy is a highly conserved degradation process that is involved in the clearance of proteins and damaged organelles to maintain intracellular homeostasis and cell integrity. Type 2 diabetes is often accompanied by dyslipidemia with elevated levels of free fatty acids (FFAs). Podocytes, as an important component of the filtration , are susceptible to lipid disorders. The loss of podocytes causes proteinuria, which is involved in the pathogenesis of diabetic nephropathy. In the present study, we demonstrated that (PA) promoted autophagy in podocytes. We further found that PA increased the production of reactive oxygen species (ROS) in podocytes and that NAC (N-acetyl-cysteine), a potent antioxidant, significantly eliminated the excessive ROS and suppressed autophagy, indicating that the increased generation of ROS was associated with the -induced autophagy in podocytes. Moreover, we also found that PA stimulation decreased the mitochondrial membrane potential in podocytes and induced podocyte apoptosis, while the inhibition of autophagy by chloroquine (CQ) enhanced -induced apoptosis accompanied by increased ROS generation, and the stimulation of autophagy by rapamycin (Rap) remarkably suppressed -induced ROS generation and apoptosis. Taken together, these in vitro findings suggest that PA-induced autophagy in podocytes is mediated by ROS production and that autophagy plays a protective role against PA-induced podocyte apoptosis.

Keyword: barrier intergrity

Kinetics of membrane raft formation: fatty domains in stratum corneum lipid models.

The major to permeability in skin resides in the outermost layer of the epidermis, the stratum corneum (SC). The major SC lipid components are ceramides, free fatty acids, and cholesterol. Ternary mixtures containing these constituents are widely used for physicochemical characterization of the . Prior X-ray diffraction and IR spectroscopy studies have revealed the existence of ordered lipid chains packed in orthorhombic subcells. To monitor the kinetics of formation of regions rich in fatty acids, the current study utilizes a modification of the method (J. Phys. Chem. 1992, 96, 10008) developed to monitor component demixing in n-alkane mixtures. The approach is based on changes in the scissoring or rocking mode contours in the IR spectra of (orthorhombically packed) ordered chains. In the current study, equimolar mixtures of ceramides (either non-hydroxy fatty sphingosine ceramide or alpha-hydroxy fatty sphingosine ceramide) with chain perdeuterated fatty acids (either or stearic ) and cholesterol reveal a time evolution of the scissoring contour of the deuterated fatty chains following quenching from relatively high temperatures where random mixing occurs. Segregation of domains enriched in the fatty component is observed. The kinetics of segregation are sensitive to the quenching temperature and to the chemical composition of the mixture. The kinetic regimes are conveniently catalogued with a power law of the form P=Ktalpha where P is a (measured) property related to domain composition. The time scales for demixing in these experiments are similar to times observed in several studies that have tracked the restoration of the in vivo permeability following nonthermal challenges to SC . Further evidence for the physiological importance of the current measurements is the detection of these phases in native SC. The current work constitutes the first direct, structure-based determination of the kinetics of formation in relevant skin lipid models.

Keyword: barrier intergrity

Effects of roasting temperature and duration on fatty composition, phenolic composition, Maillard reaction degree and antioxidant attribute of almond (Prunus dulcis) kernel.

Roasting treatment increased levels of unsaturated fatty acids (linoleic, oleic and elaidic acids) as well as saturated fatty acids ( and stearic acids) in almond (Prunus dulcis) kernel oils with temperature (150 or 180 °C) and duration (5, 10 or 20 min). Nonetheless, higher temperature (200 °C) and longer duration (10 or 20 min) roasting might result in breakdown of fatty acids especially for unsaturated fatty acids. Phenolic components (total phenols, flavonoids, condensed tannins and phenolic acids) of almond kernels substantially lost in the initial phase; afterward these components gradually increased with roasting temperature and duration. Similar results also observed for their antioxidant activities (scavenging DPPH and ABTS(+) radicals and ferric reducing power). The changes of phenolic and flavonoid compositions were also determined by HPLC. Maillard reaction products (estimated with non-enzymatic browning index) also increased with roasting temperature and duration; they might also contribute to enhancing the antioxidant attributes.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: browning

9-PAHSA promotes browning of white via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract obesity in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white . 9-PAHSA, an endogenous mammalian , which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of obesity.Copyright © 2018. Published by Elsevier Inc.

Keyword: browning

New role of irisin in hepatocytes: The protective effect of hepatic steatosis in vitro.

Irisin is a newly identified myokine related to exercise and the browning of white . Recently, it was reported that irisin serum levels are associated with intrahepatic triglyceride content, suggesting that it might have an important role in the liver. The aim of this study was to determine the role of irisin in hepatocytes. Specifically, the effect of recombinant irisin on (PA)-induced lipogenesis and its related signal pathways were examined in AML12 cells and mouse primary hepatocytes. In the present study, we observed the presence of irisin inside the cells in response to the treatment of recombinant irisin by flow cytometry and cell imaging technique. Recombinant irisin significantly inhibited the PA-induced increase in lipogenic markers ACC and FAS at the mRNA and protein levels, and prevented the PA-induced accumulation in hepatocytes. Additionally, irisin inhibited the PA-induced increase in the expression, nuclear localization, and transcriptional activities of the master regulators of lipogenesis (LXRα and SREBP-1c). Moreover, irisin attenuated PA-induced oxidative stress, which was confirmed by measuring the expression of inflammatory markers (NFκB, COX-2, p38 MAPK, TNF, IL-6) and superoxide indicator (dihydroethidium). The preventive effects of irisin against lipogenesis and oxidative stress were mediated by the inhibition of protein arginine methyltransferase-3 (PRMT3). These findings suggested that irisin might have a beneficial role in the prevention of hepatic steatosis by altering the expression of lipogenic genes and attenuating oxidative stress in a PRMT3 dependent manner.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: browning

Myricanol modulates skeletal muscle-adipose tissue crosstalk to alleviate high-fat diet-induced obesity and insulin resistance.

Skeletal muscle is the predominant site for glucose disposal and fatty consumption. Emerging evidence indicates that the crosstalk between adipose tissue and skeletal muscle is critical in maintaining insulin sensitivity and lipid homeostasis. The current study was designed to investigate whether myricanol improves insulin sensitivity and alleviates adiposity through modulating skeletal muscle-adipose tissue crosstalk.The therapeutic effect of myricanol was evaluated on (PA)-treated C2C12 myotubes and high-fat diet (HFD)-fed mice. The crosstalk between myotubes and adipocytes was evaluated using Transwell assay. The cellular lipid content was examined by Nile red staining. The mitochondrial content was assessed by MitoTracker Green staining and citrate synthase activity, and the mitochondrial function was examined by Seahorse assay. Expression of mitochondria-related and insulin signalling pathway proteins was analysed by Western blot, and the irisin level was determined by elisa kit.Myricanol increased mitochondrial quantity and function through activating AMP-activated protein kinase, resulting in reduced lipid accumulation and enhanced insulin-stimulated glucose uptake, in PA-treated C2C12 myotubes. Furthermore, myricanol stimulated irisin production and secretion from myotubes to reduce lipid content in 3T3-L1 adipocytes. In HFD-fed mice, myricanol treatment alleviated adiposity and insulin resistance through enhancing lipid utilization and irisin production in skeletal muscle and inducing of inguinal fat.Myricanol modulates skeletal muscle-adipose tissue crosstalk, to stimulate of adipose tissue and improve insulin sensitivity in skeletal muscle. Myricanol might be a potential candidate for treating insulin resistance and obesity.© 2019 The British Pharmacological Society.

Keyword: browning

Novel formulation of solid lipid microparticles of curcumin for anti-angiogenic and anti-inflammatory activity for optimization of therapy of inflammatory bowel disease.

This project was undertaken with a view to optimize the treatment of inflammatory bowel disease through a novel drug delivery approach for localized treatment in the colon. Curcumin has poor aqueous solubility, poor stability in the gastrointestinal tract and poor bioavailability. The purpose of the study was to prepare and evaluate the anti-inflammatory activity of solid lipid microparticles (SLMs) of curcumin for the treatment of inflammatory bowel disease in a -induced rat model by a colon-specific delivery approach.We have developed a novel formulation approach for treating experimental in the rat model. SLMs of curcumin were prepared with various lipids, such as , stearic and soya lecithin, with an optimized percentage of poloxamer 188. The SLMs of curcumin were characterized for particle size, drug content, drug entrapment, in-vitro release, surface morphology and infrared, differential scanning calorimetry and X-ray studies. The colonic delivery system of SLM formulations of curcumin were further investigated for their anti-angiogenic and anti-inflammatory activity using chick embryo and rat models.Particle size, drug content, drug entrapment and in-vitro release studies showed that formulation F4 containing one part stearic and 0.5% surfactant had the smallest diameter of 108 microm, 79.24% entrapment and exhibited excellent in-vitro release characteristics when compared with other formulations and pure curcumin. SLMs of curcumin (F4) proved to be a potent angio-inhibitory compound, as demonstrated by inhibition of angiogenesis in the chorioallantoic membrane assay. Rats treated with curcumin and its SLM complex showed a faster weight gain compared with dextran sulfate solution (DSS) control rats. The increase in whole colon length appeared to be significantly greater in SLM-treated rats when compared with pure curcumin and DSS control rats. An additional finding in the DSS-treated rats was chronic cell infiltration with predominance of eosinophils. Decreased mast cell numbers in the mucosa of the colon of SLMs of curcumin and pure curcumin-treated rats was observed.The degree of caused by administration of DSS was significantly attenuated by colonic delivery of SLMs of curcumin. Being a nontoxic natural dietary product, curcumin could be useful in the therapeutic strategy for inflammatory bowel disease patients.

Keyword: colitis

Palmitoylethanolamide normalizes intestinal motility in a model of post-inflammatory accelerated transit: involvement of CB₁ receptors and TRPV1 channels.

Palmitoylethanolamide (PEA), a naturally occurring acylethanolamide chemically related to the endocannabinoid anandamide, interacts with targets that have been identified in peripheral nerves controlling gastrointestinal motility, such as cannabinoid CB1 and CB2 receptors, TRPV1 channels and PPARα. Here, we investigated the effect of PEA in a mouse model of functional accelerated transit which persists after the resolution of colonic inflammation (post-inflammatory irritable bowel syndrome).Intestinal inflammation was induced by intracolonic administration of oil of mustard (OM). Mice were tested for motility and biochemical and molecular biology changes 4 weeks later. PEA, oleoylethanolamide and endocannabinoid levels were measured by liquid chromatography-mass spectrometry and receptor and enzyme mRNA expression by qRT-PCR.OM induced transient and a functional post-inflammatory increase in upper gastrointestinal transit, associated with increased intestinal anandamide (but not 2-arachidonoylglycerol, PEA or oleoylethanolamide) levels and down-regulation of mRNA for TRPV1 channels. Exogenous PEA inhibited the OM-induced increase in transit and tended to increase anandamide levels. had a weaker effect on transit. Inhibition of transit by PEA was blocked by rimonabant (CB1 receptor antagonist), further increased by 5'-iodoresiniferatoxin (TRPV1 antagonist) and not significantly modified by the PPARα antagonist GW6471.Intestinal endocannabinoids and TRPV1 channel were dysregulated in a functional model of accelerated transit exhibiting aspects of post-inflammatory irritable bowel syndrome. PEA counteracted the accelerated transit, the effect being mediated by CB1 receptors (possibly via increased anandamide levels) and modulated by TRPV1 channels.© 2014 The British Pharmacological Society.

Keyword: colitis

Branched Fatty Esters of Hydroxy Fatty Acids (FAHFAs) Protect against by Regulating Gut Innate and Adaptive Immune Responses.

We recently discovered a structurally novel class of endogenous lipids, branched esters of hydroxy stearic acids (PAHSAs), with beneficial metabolic and anti-inflammatory effects. We tested whether PAHSAs protect against , which is a chronic inflammatory disease driven predominantly by defects in the innate mucosal barrier and adaptive immune system. There is an unmet clinical need for safe and well tolerated oral therapeutics with direct anti-inflammatory effects. Wild-type mice were pretreated orally with vehicle or 5-PAHSA (10 mg/kg) and 9-PAHSA (5 mg/kg) once daily for 3 days, followed by 10 days of either 0% or 2% dextran sulfate sodium water with continued vehicle or PAHSA treatment. The colon was collected for histopathology, gene expression, and flow cytometry. Intestinal crypt fractions were prepared for ex vivo bactericidal assays. Bone marrow-derived dendritic cells pretreated with vehicle or PAHSA and splenic CD4 T cells from syngeneic mice were co-cultured to assess antigen presentation and T cell activation in response to LPS. PAHSA treatment prevented weight loss, improved scores (stool consistency, hematochezia, and mouse appearance), and augmented intestinal crypt Paneth cell bactericidal potency via a mechanism that may involve GPR120. In vitro, PAHSAs attenuated dendritic cell activation and subsequent T cell proliferation and Th1 polarization. The anti-inflammatory effects of PAHSAs in vivo resulted in reduced colonic T cell activation and pro-inflammatory cytokine and chemokine expression. These anti-inflammatory effects appear to be partially GPR120-dependent. We conclude that PAHSA treatment regulates innate and adaptive immune responses to prevent mucosal damage and protect against . Thus, PAHSAs may be a novel treatment for and related inflammation-driven diseases.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: colitis

Palmatine attenuated dextran sulfate sodium (DSS)-induced via promoting mitophagy-mediated NLRP3 inflammasome inactivation.

Activation of NLRP3 inflammasomes is crucial in the pathological process of Ulcerative (UC), which could be negatively regulated by PINK1/Parkin-driven mitophagy. Palmatine is a herb derived isoquinoline alkaloid with potent anti-inflammatory and anti-bacteria activities. In present study, we evaluated the effect of palmatine on dextran sulfate sodium (DSS)-induced mice and examined whether its effect is exerted by promoting mitophagy-mediated NLRP3 inflammasome inactivation. The result showed that palmatine (40, 100\u2009mg/kg) significantly prevented bodyweight loss and colonic shortening in DSS mice, and reduced the disease activity index and histopathologic score. The levels of MPO, IL-1β, TNF-α and the number of F4/80+ cells in colon of DSS mice were remarkably decreased by palmatine. Moreover, palmatine suppressed NLRP3 inflammasomes activation, but enhanced the expression of the mitophagy-related proteins involving LC3, PINK1 and Parkin in colonic tissue of DSS mice. These effects was consistent with the in vitro data revealing that palmatine inhibited the activation of NLRP3 inflammasomes, while promoted the expression and mitochondrial recruitment of PINK1 and Parkin in THP-1 cell differentiated macrophages. Furthermore, the effect of palmatine on THP-1 cells was neutralized by a mitophagy inhibitor Cyclosporin A (CsA) and PINK1-siRNA. In parallel, CsA significantly attenuated the therapeutic effect of palmatine in DSS mice, illustrating that the anti- effect of palmatine is closely related to mitophagy. Taken together, the current results demonstrated that palmatine protected mice against DSS-induced by facilitating PINK1/Parkin-driven mitophagy and thus inactivating NLRP3 inflammasomes in macrophage.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: colitis

High beta-palmitate fat controls the intestinal inflammatory response and limits intestinal damage in mucin Muc2 deficient mice.

esterified to the sn-1,3 positions of the glycerol backbone (alpha, alpha'-palmitate), the predominant palmitate conformation in regular infant formula fat, is poorly absorbed and might cause abdominal discomfort. In contrast, esterified to the sn-2 position (beta-palmitate), the main palmitate conformation in human milk fat, is well absorbed. The aim of the present study was to examine the influence of high alpha, alpha'-palmitate fat (HAPF) diet and high beta-palmitate fat (HBPF) diet on development in Muc2 deficient (Muc2(-/-)) mice, a well-described animal model for spontaneous enterocolitis due to the lack of a protective mucus layer.Muc2(-/-) mice received AIN-93G reference diet, HAPF diet or HBPF diet for 5 weeks after weaning. Clinical symptoms, intestinal morphology and inflammation in the distal colon were analyzed.Both HBPF diet and AIN-93G diet limited the extent of intestinal erosions and morphological damage in Muc2(-/-) mice compared with HAPF diet. In addition, the immunosuppressive regulatory T (Treg) cell response as demonstrated by the up-regulation of Foxp3, Tgfb1 and Ebi3 gene expression levels was enhanced by HBPF diet compared with AIN-93G and HAPF diets. HBPF diet also increased the gene expression of Pparg and enzymatic antioxidants (Sod1, Sod3 and Gpx1), genes all reported to be involved in promoting an immunosuppressive Treg cell response and to protect against .This study shows for the first time that HBPF diet limits the intestinal mucosal damage and controls the inflammatory response in Muc2(-/-) mice by inducing an immunosuppressive Treg cell response.

Keyword: colitis

Palmitoylethanolamide, a naturally occurring lipid, is an orally effective intestinal anti-inflammatory agent.

Palmitoylethanolamide (PEA) acts via several targets, including cannabinoid CB1 and CB2 receptors, transient receptor potential vanilloid type-1 (TRPV1) ion channels, peroxisome proliferator-activated receptor alpha (PPAR α) and orphan G protein-coupled receptor 55 (GRR55), all involved in the control of intestinal inflammation. Here, we investigated the effect of PEA in a murine model of . was induced in mice by intracolonic administration of dinitrobenzenesulfonic (DNBS). Inflammation was assessed by evaluating inflammatory markers/parameters and by histology; intestinal permeability by a fluorescent method; colonic cell proliferation by immunohistochemistry; PEA and endocannabinoid levels by liquid chromatography mass spectrometry; receptor and enzyme mRNA expression by quantitative RT-PCR.DNBS administration caused inflammatory damage, increased colonic levels of PEA and endocannabinoids, down-regulation of mRNA for TRPV1 and GPR55 but no changes in mRNA for CB1 , CB2 and PPARα. Exogenous PEA (i.p. and/or p.o., 1\u2009mg·kg(-1) ) attenuated inflammation and intestinal permeability, stimulated colonic cell proliferation, and increased colonic TRPV1 and CB1 receptor expression. The anti-inflammatory effect of PEA was attenuated or abolished by CB2 receptor, GPR55 or PPARα antagonists and further increased by the TRPV1 antagonist capsazepine.PEA improves murine experimental , the effect being mediated by CB2 receptors, GPR55 and PPARα, and modulated by TRPV1 channels.© 2014 The British Pharmacological Society.

Keyword: colitis

Increased arachidonic levels in phospholipids of human colonic mucosa in inflammatory bowel disease.

1. Colonic mucosa from 19 patients with ulcerative , eight with Crohn's disease and 14 controls were analysed for arachidonic (C20:4), linoleic (C18:2), oleic (C18:1), stearic (C18:0) and (C16:0). 2. Gas-liquid chromatography of lipid extracts showed that arachidonic was significantly higher in ulcerative (19 +/- 4) and Crohn's disease (20 +/- 3) than in controls (13 +/- 5 micrograms/mg of protein) (means +/- SD). Neither the degree of inflammation nor treatment with sulphasalazine or prednisolone appeared to influence the fatty concentrations. 3. Seventy-five to ninety-five per cent of the arachidonic was found in the phospholipid fraction after separation by thin-layer chromatography. There were no significant changes in the concentrations of the other fatty acids measured, although oleic was lower in inflammatory bowel disease. The ratios of oleic to stearic and to were lower in inflammatory bowel disease. 4. The alteration in the fatty profile may partly explain the increased synthesis of eicosanoids in colonic mucosa in inflammatory bowel disease.

Keyword: colitis

Adelmidrol, a Palmitoylethanolamide Analogue, as a New Pharmacological Treatment for the Management of Inflammatory Bowel Disease.

Leukocyte infiltration, improved levels of intercellular adhesion molecule 1 (ICAM-1), and oxidative stress in the colon are the principal factors in inflammatory bowel disease. The goal of the current study was to explore the effects of adelmidrol, an analog of the anti-inflammatory fatty amide signaling molecule palmitoylethanolamide, in mice subjected to experimental . Additionally, to clarify whether the protective action of adelmidrol is dependent on the activation of peroxisome proliferator-activated receptors (PPARs), we investigated the effects of a PPARγ antagonist, GW9662, on adelmidrol action. Adelmidrol (10 mg/kg daily, o.s.) was tested in a murine experimental model of induced by intracolonic administration of dinitrobenzene sulfonic . Nuclear factor-κB translocation, cyclooxygenase-2, and phosphoextracellular signal-regulated kinase, as well as tumor necrosis factor-α and interleukin-1β, were significantly increased in colon tissues after dinitrobenzene sulfonic administration. Immunohistochemical staining for ICAM-1, P-selectin, nitrotyrosine, and poly(ADP)ribose showed a positive staining in the inflamed colon. Treatment with adelmidrol decreased diarrhea, body weight loss, and myeloperoxidase activity. Adelmidrol treatment, moreover, reduced nuclear factor-κB translocation, cyclooxygenase-2, and phosphoextracellular signal-regulated kinase expression; proinflammatory cytokine release; and the incidence of nitrotyrosine and poly(ADP)ribose in the colon. It also decreased the upregulation of ICAM-1 and P-selectin. Adelmidrol treatment produced a reduction of Bax and an intensification of Bcl-2 expression. This study clearly demonstrates that adelmidrol exerts important anti-inflammatory effects that are partly dependent on PPARγ, suggesting that this molecule may represent a new pharmacologic approach for inflammatory bowel disease treatment.Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: colitis

Ethanol fraction of Aralia elata Seemann enhances antioxidant activity and lowers serum lipids in rats when administered with benzo(a)pyrene.

Aralia elata Seemann is an edible mountain vegetable in Korea containing saponin, alkaloid, , linoleic , methyl eicosanoate and hexacosol, and is known to manifest an effect on cardiac infarction, gastric ulcer, , and enervation. This study has examined the effects of Aralia elata Seemann ethanol extract on antioxidant enzyme systems and lipid metabolism in rats along with benzo(a)pyrene (B(a)P) administration. Rats were divided into four groups: control (C), an extract fed group (CE), a B(a)P fed group (CB), and a B(a)P and extract fed group (CBE). The ethanol extracts of Aralia elata Seemann (50 mg/kg body weight) were fed to the rats for 4 weeks by stomach tubing. Extract administration increased the antioxidant activities of glutathione sulfur transferase (GST). Total superoxide dismutase (SOD) and Cu,Zn-SOD activities were stimulated. Catalase activities were increased by 50% with extract feeding. Cu,Zn-SOD was greatly enhanced from 0.10 unit to 0.18 unit and catalase activity also was increased. Serum alpha-tocopherol was markedly increased by the extracts. The ethanol fraction of Aralia elata Seemann decreased total serum cholesterol. However, serum HDL-cholesterol was increased by 35% (p<0.05). The results indicate that Aralia elata Seemann exerts antioxidant and strong hypocholesterolemic and hypolipidemic effects in vivo with the administration of B(a)P.

Keyword: colitis

Modulation in delta 9, delta 6, and delta 5 fatty desaturase activity in the human intestinal CaCo-2 cell line.

We report the influence of media lipids, growth in lipid-poor medium, and cell differentiation on delta 9, delta 6, and delta 5 desaturase activity in the human CaCo-2 enterocyte cell line. We also describe the level of incorporation of (16:0), linoleic (18:2n-6), and eicosapentaenoic (EPA) acids (20:5n-3) and their higher homologues into cytosolic and membrane lipids during long-term (10 days) medium supplementation in fully differentiated 16- to 18-day-old cultures. CaCo-2 monolayers reached confluency by day 6 with subsequent development of microvilli and maximal expression of microvillus membrane sucrose, alkaline phosphatase, and gamma-glutamyltransaminase occurring between days 16 and 23 after plating. There was evidence of the presence and modulation of delta 9, delta 6, and delta 5 desaturase activity (delta 9 > delta 6 > delta 5). delta 6 Desaturase activity decreased approximately 2-fold between days 6 and 24 of culture and when the fetal bovine serum concentration was increased from 0.5% to 25%; in contrast, when cells were starved for 72 h, activity increased 5.4-fold. When the media was supplemented with either linoleic and/or EPA, both delta 6 and delta 5 desaturase activities were inhibited, the greatest reduction of delta 5 desaturase activity occurring with EPA. Incorporation of media fatty acids plus their desaturase and elongase products was highly dependent on medium composition with the homologues of delta 9 > delta 6 > delta 5. Supplementation of cellular media with 100 microM EPA for 10 days decreased membrane phosphatidylethanolamine arachidonic level from 13.2 to 8.9%. From these results we conclude that enterocyte membrane fatty composition and desaturase enzyme activity are regulated by both dietary fat intake and cell maturation. The clinical relevance of these observations on lipid dietary modification for the management of chronic inflammatory bowel disease is still uncertain but these observations suggest that the beneficial effects of EPA supplements on human ulcerative may be due to a reduction in enterocyte arachidonic content by down-regulation of delta 6 and delta 5 desaturase activity.

Keyword: colitis

Docosahexaenoyl serotonin emerges as most potent inhibitor of IL-17 and CCL-20 released by blood mononuclear cells from a series of N-acyl serotonins identified in human intestinal tissue.

Fatty amides (FAAs), conjugates of fatty acids with ethanolamine, mono-amine neurotransmitters or amino acids are a class of molecules that display diverse functional roles in different cells and tissues. Recently we reported that one of the serotonin-fatty conjugates, docosahexaenoyl serotonin (DHA-5-HT), previously found in gut tissue of mouse and pig, attenuates the IL-23-IL-17 signaling axis in LPS-stimulated mice macrophages. However, its presence and effects in humans remained to be elucidated. Here, we report for the first time its identification in human intestinal (colon) tissue, along with a series of related N-acyl serotonins. Furthermore, we tested these fatty conjugates for their ability to inhibit the release of IL-17 and CCL-20 by stimulated human peripheral blood mononuclear cells (PBMCs). Serotonin conjugates with (PA-5-HT), stearic (SA-5-HT) and oleic (OA-5-HT) were detected in higher levels than arachidonoyl serotonin (AA-5-HT) and DHA-5-HT, while eicosapentaenoyl serotonin (EPA-5-HT) could not be quantified. Among these, DHA-5-HT was the most potent in inhibiting IL-17 and CCL-20, typical Th17 pro-inflammatory mediators, by Concanavalin A (ConA)-stimulated human PBMCs. These results underline the idea that DHA-5-HT is a gut-specific endogenously produced mediator with the capacity to modulate the IL-17/Th17 signaling response. Our findings may be of relevance in relation to intestinal inflammatory diseases like Crohn's disease and Ulcerative .Copyright © 2017. Published by Elsevier B.V.

Keyword: colitis

The ether lipid precursor hexadecylglycerol protects against Shiga toxins.

Shiga toxin-producing Escherichia coli bacteria cause hemorrhagic and hemolytic uremic syndrome in humans. Currently, only supportive treatment is available for diagnosed patients. We show here that 24-h pretreatment with an ether lipid precursor, the alkylglycerol sn-1-O-hexadecylglycerol (HG), protects HEp-2 cells against Shiga toxin and Shiga toxin 2. Also the endothelial cell lines HMEC-1 and HBMEC are protected against Shiga toxins after HG pretreatment. In contrast, the corresponding acylglycerol, DL-α-palmitin, has no effect on Shiga toxicity. Although HG treatment provides a strong protection (~30 times higher IC₅₀) against Shiga toxin, only a moderate reduction in toxin binding was observed, suggesting that retrograde transport of the toxin from the plasma membrane to the cytosol is perturbed. Furthermore, endocytosis of Shiga toxin and retrograde sorting from endosomes to the Golgi apparatus remain intact, but transport from the Golgi to the endoplasmic reticulum is inhibited by HG treatment. As previously described, HG reduces the total level of all quantified glycosphingolipids to 50-70% of control, including the Shiga toxin receptor globotriaosylceramide (Gb3), in HEp-2 cells. In accordance with this, we find that interfering with Gb3 biosynthesis by siRNA-mediated knockdown of Gb3 synthase for 24 h causes a similar cytotoxic protection and only a moderate reduction in toxin binding (to 70% of control cells). Alkylglycerols, including HG, have been administered to humans for investigation of therapeutic roles in disorders where ether lipid biosynthesis is deficient, as well as in cancer therapy. Further studies may reveal if HG can also have a therapeutic potential in Shiga toxin-producing E. coli infections.

Keyword: colitis

N-Acylethanolamine-hydrolyzing amidase inhibition increases colon N-palmitoylethanolamine levels and counteracts murine .

N-Palmitoylethanolamine or palmitoylethanolamide (PEA) is an anti-inflammatory compound that was recently shown to exert peroxisome proliferator-activated receptor-α-dependent beneficial effects on colon inflammation. The actions of PEA are terminated following hydrolysis by 2 enzymes: fatty amide hydrolase (FAAH), and the less-studied N-acylethanolamine-hydrolyzing amidase (NAAA). This study aims to investigate the effects of inhibiting the enzymes responsible for PEA hydrolysis in colon inflammation in order to propose a potential therapeutic target for inflammatory bowel diseases (IBDs). Two murine models of IBD were used to assess the effects of NAAA inhibition, FAAH inhibition, and PEA on macroscopic signs of colon inflammation, macrophage/neutrophil infiltration, and the expression of proinflammatory mediators in the colon, as well as on the -related systemic inflammation. NAAA inhibition increases PEA levels in the colon and reduces colon inflammation and systemic inflammation, similarly to PEA. FAAH inhibition, however, does not increase PEA levels in the colon and does not affect the macroscopic signs of colon inflammation or immune cell infiltration. This is the first report of an anti-inflammatory effect of a systemically administered NAAA inhibitor. Because NAAA is the enzyme responsible for the control of PEA levels in the colon, we put forth this enzyme as a potential therapeutic target in chronic inflammation in general and IBD in particular.© FASEB.

Keyword: colitis

High-fat diet promotes experimental by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) tight junction regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes in a 2,4,6-trinitrobenzenesulfonic -induced experimental model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: colitis

The influence of the position of palmitate in infant formula triacylglycerols on health outcomes.

The purpose of this review is to discuss recent studies reporting on the influence of the position of in triacylglycerols in infant formula and relevant animal studies. Earlier experiments in rodents show that a diet with a higher proportion of palmitate at the sn-2 position of triacylglycerols improves dietary fat and calcium absorption compared with a diet with a lower sn-2 palmitate content. A high-sn-2 palmitate diet increased fecal short-chain fatty acids, reduced gut inflammation in a model, and altered tissue endocannabinoid concentrations in laboratory rodents. Recent studies in infants confirm that formula with a high sn-2 palmitate content reduces stool fat, , fat soaps, palmitate soaps, and calcium compared with formula with a low sn-2 palmitate content. These effects have been associated with improved bone strength, increased fecal bifidobacteria, and reduced crying in infants. In some studies, findings with formula high in sn-2 palmitate match those seen in breast-fed infants. However, in many studies, high sn-2 palmitate formula remains inferior to breast-feeding. It is concluded that infant formula high in sn-2 palmitate is superior to formula with low sn-2 palmitate but does not fully match human breast milk. Recent studies showing altered gut microbiota (human infants) and tissue endocannabinoids (rodent model) suggest the potential for marked physiological impact of high sn-2 palmitate that needs to be explored further in human trials.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: colitis

Preventive and curative effect of Pistacia lentiscus oil in experimental .

to investigate the anti-inflammatory effect of the Pistacia lentiscus oil in experimental model. was induced in male rats by instillation of 2,4,6-trinitrobenzenesulfonic (TNBS) in all groups. The experimental groups consisted of: 5 rats received Lentisc oil 2months before induction (preventive group), 5 rats received the oil on the day of induction (curative group) and 5 control rats. Lentisc oil was extracted from the ripe fruit of the plant by the cold press method and was analyzed by spectro-chromatography. Lentisc oil has been inserted with a standard diet at the dose of 30mg oil/100g of food/rat.The lentisc oil sample is composed mainly by Oleic (47.96%), (27.94%) and Linoleic (20.22%).There was a significant difference between control rats and treated rats with lentisc oil concerned body mass (p=0.009), bleeding index (p=0.005 and p=0.018) and diarrhea (p=0.012). Histological examination revealed a clear difference between the control and preventive groups with disappearance of erosion, decreased of cryptitis, irregular crypts and crypt loss in the preventive group. Curative group showed a significant decrease of ulceration, hyperplasia, cryptitis, irregular crypts and crypt loss compared to the control group. There was an attenuation of inflammation in the preventive group compared to the curative group without statistically significant.Lentisc oil administration could provide a protective effect on intestinal inflammation in rats induced by TNBS mainly when it is administered at a young age in preventive mode. This beneficial effect would involve a modification of arachidonic metabolism.Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Keyword: colitis

Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental .

Autophagy, which is critical for the proper turnover of organelles such as endoplasmic reticulum and mitochondria, affects diverse aspects of metabolism, and its dysregulation has been incriminated in various metabolic disorders. However, the role of autophagy of myeloid cells in adipose tissue inflammation and type 2 diabetes has not been addressed. We produced mice with myeloid cell-specific deletion of Atg7 (autophagy-related 7), an essential autophagy gene (Atg7 conditional knockout [cKO] mice). While Atg7 cKO mice were metabolically indistinguishable from control mice, they developed diabetes when bred to ob/w mice (Atg7 cKO-ob/ob mice), accompanied by increases in the crown-like structure, inflammatory cytokine expression and inflammasome activation in adipose tissue. Mφs (macrophages) from Atg7 cKO mice showed significantly higher interleukin 1 β release and inflammasome activation in response to a plus lipopolysaccharide combination. Moreover, a decrease in the NAD(+):NADH ratio and increase in intracellular ROS content after treatment with in combination with lipopolysaccharide were more pronounced in Mφs from Atg7 cKO mice, suggesting that mitochondrial dysfunction in autophagy-deficient Mφs leads to an increase in lipid-induced inflammasome and metabolic deterioration in Atg7 cKO-ob/ob mice. Atg7 cKO mice were more susceptible to experimental , accompanied by increased colonic cytokine expression, T helper 1 skewing and systemic bacterial invasion. These results suggest that autophagy of Mφs is important for the control of inflammasome activation in response to metabolic or extrinsic stress, and autophagy deficiency in Mφs may contribute to the progression of metabolic syndrome associated with lipid injury and .

Keyword: colitis

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Inhibition of antibacterial activity of himastatin, a new antitumor antibiotic from Streptomyces hygroscopicus, by fatty sodium salts.

Himastatin, a cyclohexadepsipeptide antibiotic, had in vivo antitumor activity against localized P388 leukemia and B16 melanoma but had no distal site antitumor activity. An in vitro Bacillus subtilis well-agar diffusion assay was employed to test the hypothesis that himastatin was enzymatically inactivated. The activity of himastatin against B. subtilis was inhibited when himastatin was mixed with mouse liver S9 fraction and microsomes. However, subsequent investigations demonstrated that the markedly decreased antibacterial activity was not enzymatic in nature but was related to the presence of certain fatty salts. Saturated fatty sodium salts with a carbon chain number of 8 or more reduced the antimicrobial activity of himastatin 50 to 100 times. If antibiotics such as ampicillin, bacitracin, chloramphenicol, and tunicamycin were used in place of himastatin, no meaningful reduction in antibacterial activity occurred. However, the antibacterial activity of the membrane-active peptide antibiotic polymyxin B, but not that of polymyxin E (colistin), was reduced in a manner similar to that of himastatin. Importantly, the activity of himastatin against HCT-116 adenocarcinoma cells in soft agar was markedly reduced in the presence of sodium palmitate as the reference fatty salt. The data indicate that himastatin may be trapped in micelles in vitro. It may be speculated that the lack of distal site antitumor activity resulted from similar complex formation between himastatin and lipids in vivo. The results also suggest that the cytotoxic and antimicrobial effects of himastatin may result from interactions with the cell membrane.

Keyword: colon cancer

Fatty composition and anticancer activity in carcinoma cell lines of Prunus dulcis seed oil.

Almond oil is used in traditional and complementary therapies for its numerous health benefits due to high unsaturated fatty acids content.This study investigated the composition and in vitro anticancer activity of almond oil from Northern Cyprus and compared with almond oil from Turkey.Almond oil from Northern Cyprus was obtained by supercritical CO extraction and analyzed by GC-MS. Almond oil of Turkey was provided from Turkish pharmacies. Different concentrations of almond oils were incubated for 24 and 48\u2009h with Colo-320 and Colo-741 cells. Cell growth and cytotoxicity were measured by MTT assays. Anticancer and antiprolifetarive activities of almond oils were investigated by immunocytochemistry using antibodies directed against to BMP-2, β-catenin, Ki-67, LGR-5 and Jagged 1.Oleic (77.8%; 75.3%), linoleic (13.5%; 15.8%), (7.4%; 6.3%), were determined as the major compounds of almond oil from Northern Cyprus and Turkey, respectively. In the MTT assay, both almond oils were found to be active against Colo-320 and Colo-741 cells with 1:1 dilution for both 24\u2009h and 48\u2009h. As a result of immunohistochemical staining, while both almond oils exhibited significant antiproliferative and anticancer activity, these activities were more similar in Colo-320 cells which were treated with Northern Cyprus almond oil.Almond oil from Northern Cyprus and Turkey may have anticancer and antiproliferative effects on cells through molecular signalling pathways and, thus, they could be potential novel therapeutic agents.

Keyword: colon cancer

5'-O-Palmitoyl- and 3',5'-O-dipalmitoyl-5-fluoro-2'-deoxyuridine--novel lipophilic analogues of 5'-fluoro-2'-deoxyuridine: synthesis, incorporation into liposomes and preliminary biological results.

5'-O-palmitoyl- and 3',5'-O-dipalmitoyl-5-fluoro-2'-deoxyuridine were prepared by the reaction of 5-fluoro-2'-deoxyuridine in dimethylacetamide with chloride. The incorporation of the synthesized prodrugs into liposomes composed of egg phosphatidylcholine/stearylamine/cholesterol/alpha-tocopherol at a molar ratio of 10:1:2:0.05 was nearly quantitative; homogeneous bilayer vesicles (75 nm diameter) were obtained. Preliminary tolerance studies revealed that the prodrug-liposome preparations are about 20-60 times more toxic than the parent drug. The prodrugs incorporated into liposomes were 10 to 30 times more active against murine 38 carcinoma compared to the free drug. In comparison to the administration of the prodrugs in peanut oil the liposomal preparations seem to exert improved effects and represent a valuable drug delivery system for parenteral applications.

Keyword: colon cancer

Myristoylation of heparan sulfate proteoglycan and proteins occurs post-translationally in human carcinoma cells.

We have recently shown that the heparan sulfate proteoglycan of human carcinoma cells is acylated with both myristate and palmitate, two long-chain saturated fatty acids. In this study we show that cycloheximide did not significantly inhibit the incorporation of myristic into either proteoglycan or total protein pool. This lack of inhibition occurred under a condition in which protein synthesis was inhibited greater than 90%. Cycloheximide, on the other hand, did not affect the incorporation of [3H]myristic into fatty nor the intracellular interconversion of myristate to palmitate. Characterization of fatty acyl moiety in the proteoglycan and protein by reverse-phase HPLC revealed that approximately 60% of the covalently bound fatty acids was myristate and the remaining 40% was palmitate. These results indicate that in human carcinoma cells myristoylation of heparan sulfate proteoglycan and proteins occurs post-translationally, presumably in the Golgi complex.

Keyword: colon cancer

Aldo-keto reductase family 1 member B10 promotes cell survival by regulating lipid synthesis and eliminating carbonyls.

Aldo-keto reductase family 1 member B10 (AKR1B10) is primarily expressed in the normal human and small intestine but overexpressed in liver and lung . Our previous studies have shown that AKR1B10 mediates the ubiquitin-dependent degradation of acetyl-CoA carboxylase-alpha. In this study, we demonstrate that AKR1B10 is critical to cell survival. In human carcinoma cells (HCT-8) and lung carcinoma cells (NCI-H460), small-interfering RNA-induced AKR1B10 silencing resulted in caspase-3-mediated apoptosis. In these cells, the total and subspecies of cellular lipids, particularly of phospholipids, were decreased by more than 50%, concomitant with 2-3-fold increase in reactive oxygen species, mitochondrial cytochrome c efflux, and caspase-3 cleavage. AKR1B10 silencing also increased the levels of alpha,beta-unsaturated carbonyls, leading to the 2-3-fold increase of cellular lipid peroxides. Supplementing the HCT-8 cells with (80 mum), the end product of fatty synthesis, partially rescued the apoptosis induced by AKR1B10 silencing, whereas exposing the HCT-8 cells to epalrestat, an AKR1B10 inhibitor, led to more than 2-fold elevation of the intracellular lipid peroxides, resulting in apoptosis. These data suggest that AKR1B10 affects cell survival through modulating lipid synthesis, mitochondrial function, and oxidative status, as well as carbonyl levels, being an important cell survival protein.

Keyword: colon cancer

Human carbonyl reductase 1 participating in intestinal first-pass drug metabolism is inhibited by fatty acids and acyl-CoAs.

Human carbonyl reductase 1 (CBR1), a member of the short-chain dehydrogenase/reductase (SDR) superfamily, reduces a variety of carbonyl compounds including endogenous isatin, prostaglandin E and 4-oxo-2-nonenal. It is also a major non-cytochrome P450 enzyme in the phase I metabolism of carbonyl-containing drugs, and is highly expressed in the intestine. In this study, we found that long-chain fatty acids and their CoA ester derivatives inhibit CBR1. Among saturated fatty acids, myristic, and stearic acids were inhibitory, and stearic was the most potent (IC 9µM). Unsaturated fatty acids (oleic, elaidic, γ-linolenic and docosahexaenoic acids) and acyl-CoAs (palmitoyl-, stearoyl- and oleoyl-CoAs) were more potent inhibitors (IC 1.0-2.5µM), and showed high inhibitory selectivity to CBR1 over its isozyme CBR3 and other SDR superfamily enzymes (DCXR and DHRS4) with CBR activity. The inhibition by these fatty acids and acyl-CoAs was competitive with respect to the substrate, showing the K values of 0.49-1.2µM. Site-directed mutagenesis of the substrate-binding residues of CBR1 suggested that the interactions between the fatty acyl chain and the enzyme's Met141 and Trp229 are important for the inhibitory selectivity. We also examined CBR1 inhibition by oleic in cellular levels: The fatty effectively inhibited CBR1-mediated 4-oxo-2-nonenal metabolism in DLD1 cells and increased sensitivity to doxorubicin in the drug-resistant gastric MKN45 cells that highly express CBR1. The results suggest a possible new food-drug interaction through inhibition of CBR1-mediated intestinal first-pass drug metabolism by dietary fatty acids.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: colon cancer

Characterization of posttranslational modifications of human A33 antigen, a novel palmitoylated surface glycoprotein of human gastrointestinal epithelium.

Monoclonal antibody (mAb) A33 recognizes a differentiation antigen (A33) expressed in normal human gastrointestinal epithelium and in 95% of human cancers. Murine mAb A33 shows specific targeting of in humans and a humanized A33 antibody is currently being evaluated in the clinic. The cDNA for the human A33 antigen has recently been cloned, and sequence comparison indicated that the A33 antigen is a novel human cell surface molecule of the immunoglobulin superfamily. Because mAb A33 recognizes a conformational epitope, only a partial characterization of the A33 antigen has been carried out to date. In this report we show that the A33 antigen is (I) N-glycosylated, containing approximately 8 K of N-linked carbohydrate and there is no evidence for O-glycosylation, sialylation or glycophosphatidylinositol, and (ii) S-acylated in vitro, incorporating [3H] linked through a hydroxylamine-sensitive thioester bond. The S-palmitoylation may be involved in regulating the internalization process initiated by binding of mAb A33 to cell surface A33 antigen.

Keyword: colon cancer

In vitro anti-inflammatory and anti-proliferative activity of sulfolipids from the red alga Porphyridium cruentum.

A sulfoglycolipidic fraction (SF) isolated from the red microalga Porphyridium cruentum was analyzed for fatty composition and assayed for ability to inhibit, in vitro, the generation of superoxide anion in primed leucocytes and the proliferation of a panel of human cell-lines. Results demonstrated that SF contained large amounts of (26.1%), arachidonic (C20: 4 omega-6, 36.8%), and eicopentaenoic (C20:5 omega-3, 16.6%) acids, and noticeable amounts of 16:1n-9 fatty (10.5%). It strongly inhibited both the production of superoxide anion generated by peritoneal leukocytes primed with phorbol myristate acetate (IC(50): 29.5 microg/mL), and the growth of human adenocarcinoma DLD-1 and to a lesser extent of human breast adenocarcinoma MCF-7, human prostate adenocarcinoma PC-3, and human malignant melanoma M4 Beu cell-lines, and therefore might have a chemopreventive or chemotherapeutic potential, or both. It was found markedly more cytotoxic than sulfoquinovosyldiacylglycerols from plant used as a standard (STD), due to a stronger ability to inhibit DNA alpha-polymerase (IC(50): 378 microg/mL, vs 1784 microg/mL for STD). After a 48-h continuous treatment, IC(50) values for growth inhibition were in the range of 20-46 microg/mL instead of 94 to >250 microg/mL for STD, and those for inhibition of metabolic activity were in the range of 34-87 microg/mL instead of >250 microg/mL for STD. The higher anti-proliferative effect was observed on adenocarcinoma DLD-1 cells, and the weaker effect was observed on breast adenocarcinoma MCF-7.

Keyword: colon cancer

Antimutagenicity of yogurt.

Yogurt is milk fermented by a mixture of two bacteria: Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus. Epidemiological studies have correlated a reduced risk of with yogurt consumption. Independent studies have established that yogurt and extracts thereof are antimutagenic. Although multiple explanations can account for yogurt's putative anticarcinogenicity, we are interested in testing the hypothesis that antimutagenic compounds produced during fermentation are responsible. We recently reported on the antimutagenicity of an acetone extract of yogurt against the experimental carcinogens N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 3.2'dimethyl-4-aminobiphenyl (DMAB) (Mutation Res. (1995) 334, 213-224). We are now aware that is an active ingredient against MNNG.

Keyword: colon cancer

Comparison of phospholipid profiles of primary adenocarcinoma in the lung and other organs.

Phospholipid profiles, particularly molecular structure of phosphatidylcholine, of human primary lung adenocarcinoma were compared with those of the histologically same type of carcinoma from other organs in order to search for a possibility that differentiates between primary and metastatic tumors in the lung. The saturated class, mainly containing at both positions, accounted for 20.8% of phosphatidylcholine in lung adenocarcinoma, whereas it accounted for only 6-10.7% in the adenocarcinoma of other organs. Adenocarcinoma of organs other than the lung had specific characteristics of unsaturated molecular classes of phosphatidylcholine for each organ. Stomach tumor had high contents of dienes and tetraenes with particularly high proportion of arachidonic at the 2-position. Breast tumor had a high content of monoenes, containing palmitoleic at the 2-position. Adenocarcinomas of rectum, and thyroid contained more dienes compared to lung adenocarcinoma.

Keyword: colon cancer

Fecal diglycerides as selective endogenous mitogens for premalignant and malignant human epithelial cells.

Diglycerides (DGs) have been found in fecal extracts at concentrations which induce mitogenesis of adenoma and some carcinoma cells but not normal cells in primary culture. DGs containing stearic, oleic, , and myristic side chains were found in fecal extracts from each of eight subjects. Synthetic 1,2-DGs, containing the fatty acids found in endogenous fecal DGs, induced mitogenesis in cultures of premalignant cells from each of 13 adenomas, covering all histological classes, and in cultures from two of four carcinomas. The potent adenoma mitogen, dimyristin, had no mitogenic activity on cultures of normal epithelial cells from seven different subjects. These results suggest DGs may act as endogenous mitogens in the development of human . The extent of adenoma mitogenesis was correlated with the chain length of the saturated R-groups: 16 greater than 14 greater than 12 greater than 10 greater than 8 much greater than 18. DGs with oleic residues, C18:1, were among the most active, while substitution of even one fatty residue with a stearic residue, C18:0, reduced or eliminated mitogenic activity. Dimyristin also induced enhanced levels of urokinase secretion from carcinoma cells, in parallel to the phorbol ester tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. These results imply that DGs found in the induce a selective growth of benign tumors and some carcinomas, and may enhance the invasive capacity of carcinomas, while leaving normal cells unaffected.

Keyword: colon cancer

PKCζ Phosphorylates SIRT6 to Mediate Fatty β-Oxidation in Cells.

Protein kinase C (PKC) has critical roles in regulating lipid anabolism and catabolism. PKCζ, a member of atypical PKC family, has been reported to mediate glucose metabolism. However, whether and how PKCζ regulates tumor cells fatty β-oxidation are unknown. Here, we report that the phosphorylation of SIRT6 is significantly increased after (PA) treatment in cells. PKCζ can physically interact with SIRT6 in vitro and in vivo, and this interaction enhances following PA treatment. Further experiments show that PKCζ is the phosphorylase of SIRT6 and phosphorylates SIRT6 at threonine 294 residue to promote SIRT6 enrichment on chromatin. In the functional study, we find that the expression of ACSL1, CPT1, CACT, and HADHB, the genes related to fatty β-oxidation, increases after PA stimulation. We further confirm that PKCζ mediates the binding of SIRT6 specifically to the promoters of fatty β-oxidation-related genes and elicits the expression of these genes through SIRT6 phosphorylation. Our findings demonstrate the mechanism of PKCζ as a new phosphorylase of SIRT6 on maintaining tumor fatty β-oxidation and define the new role of PKCζ in lipid homeostasis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: colon cancer

The use of sesame oil and other vegetable oils in the inhibition of human growth in vitro.

Sesame contains large quantities of the essential polyunsaturated fatty (PUFA), linoleic , in the form triglycerides. The antineoplastic properties of many PUFAs such as linoleic and their metabolites are known. We tested the hypothesis that natural vegetable oils, such as sesame oil and its component linoleic , when added to human adenocarcinoma cells growing in tissue culture would inhibit their growth and that normal cells would not be similarly affected. Three human cell lines and one normal human cell line were exposed to the following: (1) pure linoleic ; (2) lipase-digested sesame oil; (3) undigested sesame oil; (4) five additional common vegetable oils; (5) mineral oil. Linoleic inhibited the in vitro growth of all three malignant human adenocarcinoma cell lines. The normal cell line showed dramatically less inhibition of growth. Lipase-digested sesame oil (LDSO) and undigested sesame oil (UDSO) produced greater inhibition of growth of all three malignant cell lines than of the normal cells. Five other common vegetable oils containing various amounts of PUFAs such as corn, soybean, safflower, olive and coconut oils, all in their lipase-digested form, were found to dramatically inhibit the growth of the HT-29 malignant human cell line. Undigested olive and safflower oils also inhibited the HT-29 cells although not as markedly as the lipase-digested oils. Mineral oil did not inhibit the growth of HT-29 cells. Both lauric and , which are saturated fatty acids found in abundance in coconut oil inhibits the HT-29 cells more strongly than linoleic , while oleic did not inhibit. We conclude that many vegetable oils including sesame contain in vitro antineoplastic properties and that this finding warrants further investigation both in vitro and in vivo to assess their possible chemotherapeutic potential.

Keyword: colon cancer

Role of fatty acids in malignancy and visual impairment: epidemiological evidence and experimental studies.

International variation in breast and incidence is positively related to total fat intake. However, total fat consists of different fatty families, e.g., saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and n-3 and n-6 polyunsaturated fatty acids (PUFAs). Epidemiological evidence and experimental studies suggest that these fatty families have different effects on breast and carcinogenesis. Therefore the action of each fatty on carcinogenesis should be evaluated separately. Although it is difficult to establish firm conclusions on the effect of each fatty in human epidemiological studies, experimental studies on animals and cultured cells suggest that n-6 PUFAs (linoleic and arachidonic ) may have a tumor promoting effect, while n-3 PUFAs (eicosapentaenoic , docosahexaenoic and alpha-linolenic ) and conjugated fatty acids (CFAs; a mixture of positional and geometric isomers of PUFAs with conjugated double bonds) exert an inhibitory effect on tumor growth. SFAs such as and stearic show little or no tumor promoting effect, and the action of oleic , a MUFA, is inconclusive. In addition to regulation of abnormal cell growth seen in cancers, fatty acids also control cell loss seen in degenerative eye diseases, such as degeneration of lens material in cataract and degeneration of photoreceptor cells in retinitis pigmentosa. Experiments suggest that n-6 PUFAs cause deleterious effects, while n-3 PUFAs result in beneficial effects on the lens and retina. In particular, docosahexaenoic is known to be effective in rescuing photoreceptor cells from damage. Thus, understanding the function of each fatty is likely to be important for making progress in treating these and other diseases.

Keyword: colon cancer

Type 2 Diabetes Promotes Cell Centrosome Amplification via AKT-ROS-Dependent Signalling of ROCK1 and 14-3-3σ.

Type 2 diabetes is associated with oxidative stress and DNA damage which can cause centrosome amplification. Thus, the study investigated centrosome amplification in type 2 diabetes and the underlying mechanisms.Centrosome numbers in human peripheral blood mononuclear blood cells (PBMC) from healthy subjects and patients with type 2 diabetes were compared to access the association between type 2 diabetes and centrosome amplification. cells were used to investigate the molecular mechanisms underlying the centrosome amplification triggered by high glucose, insulin and . Western blot analysis was used to quantify the level of protein and protein phosphorylation. Immunofluorescent staining was performed to detect centrosomes. ROS was quantified using flow cytometry technique. Transcriptpmic profiling was performed using Illumina HiSeqTM500 platform.We found that centrosome amplification was increased PBMC from the type 2 diabetic patients, which correlated with the levels of fasting blood glucose and HbA1c. High glucose, insulin and , alone or in combinations, induced ROS production and centrosome amplification. Together, they increased AKT activation as well as the expression, binding and centrosome translation of ROCK1 and 14-3-3σ. Results from further analyses showed that AKT-ROS-dependent upregulations of expression, binding and centrosome translocation of ROCK1 and 14-3-3σ was the molecular pathway underlying the centrosome amplification in vitro triggered by high glucose, insulin and . Moreover, the key in vitro molecular signalling events activated by high glucose, insulin and were verified in PBMC from the patients with type 2 diabetes.Our results show that type 2 diabetes promotes cell centrosome amplification, and suggest that the diabetic pathophysiological factors-activated AKT-ROS-dependent signalling of ROCK1 and 14-3-3σ is the underlying molecular mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: colon cancer

Isolation of fatty acids with anticancer activity from Protaetia brevitarsis larva.

In this study, biologically active compounds were isolated from Protaetia brevitarsis larva (PBL) by dichloromethane extraction. The dichloromethane extract from PBL was highly cytotoxic to various cells. From a silica gel column chromatograpy of this extract, we obtained four fractions (F-2, F-4, F-5 and F-7) having apoptosis-inducing activity. These fractions induced DNA ladder and caspase-3 activation during apoptosis in 26 tumor cells. In 1H and 13C NMR and mass spectral analysis of the fraction F-2 showing the highest apoptosis-inducing activity, we found that the fraction was composed of three free fatty acids such as , (Z)-9-octadecenoic and octadecenoic . These results indicate that the dichloromethane extract of PBL includes anticancer components composed of at least three fatty acids, and apoptosis-inducing activity of the extract was mediated by caspase-3 activation in tumor cells.

Keyword: colon cancer

Effects of fatty acids on the growth of Caco-2 cells.

Epidemiological studies suggest that polyunsaturated fatty acids may protect against colorectal neoplasia. In order to explore this observation, cell proliferation and viability, lipid composition, membrane fluidity, and lipid peroxidation were measured in Caco-2 cells after 48h incubation with various fatty acids. Saturated and monounsaturated fatty acids incorporated less well in the membranes than polyunsaturated fatty acids (PUFAs). All of the PUFAs tested had an inhibitory effect on cell proliferation/viability whereas the saturated and monounsaturated fatty acids did not. Addition of had no significant effect on membrane fluidity whereas unsaturated fatty acids increased membrane fluidity in a dose-dependent manner. PUFAs strongly increased tumor cell lipid peroxidation in a dose-dependent manner. Saturated and monounsaturated fatty acids increased lipid peroxidation in this cell line only at high concentration. Preincubation of Caco-2 cells with vitamin E prevented the inhibition of proliferation/viability, the elevation of the MDA concentration and the increased membrane fluidity induced by PUFAs. Our data indicate that PUFAs are potent inhibitors of the growth of cells in vitro.

Keyword: colon cancer

Mitochondrial hexokinase from differentiated and undifferentiated HT29 cells: effect of some metabolites on the bound/soluble equilibrium.

1. Solubilization of mitochondrial bound hexokinase (HK), which represents 75-80% of the total enzyme activity in the cells, was investigated in freshly isolated mitochondria from undifferentiated (Glc+) or differentiated (Glc-) HT29 adenocarcinoma cells. In both models, the bound HK is almost completely released in vitro by 100 microM glucose 6-P (G 6-P). 2. Free ATP (5 mM) or palmitate (800 microM) produce a partial solubilization of bound HK, more markedly in the case of Glc- mitochondria. 3. Glucose or glucose 1-P are found unable to solubilize bound HK. Glucose 1,6-P2, 2-deoxyglucose 6-P or glucosamine 6-P can solubilize the enzyme but are less efficient than G 6-P. 4. Mg2+ and Pi are found to counteract the glucose 6-P induced solubilization of HK in both types of mitochondria. Taking into account the intracellular concentrations of these ions, this could in part explain why, in HT29 cells, HK is predominantly bound to the mitochondria.

Keyword: colon cancer

N-(4-Hydroxyphenyl)retinamide increases dihydroceramide and synergizes with dimethylsphingosine to enhance cell killing.

Fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)] is cytotoxic in many cell types. Studies have shown that elevation of ceramide species plays a role in 4-HPR cytotoxicity. To determine 4-HPR activity in a multidrug-resistant cell line as well as to study ceramide metabolism, MCF-7/AdrR cells (redesignated NCI/ADR-RES) were treated with 4-HPR and sphingolipids were analyzed. TLC analysis of cells radiolabeled with [3H] showed that 4-HPR elicited a dose-responsive increase in radioactivity migrating in the ceramide region of the chromatogram and a decrease in cell viability. Results from liquid chromatography/electrospray tandem mass spectrometry revealed large elevations in dihydroceramides (N-acylsphinganines), but not desaturated ceramides, and large increases in complex dihydrosphingolipids (dihydrosphingomyelins, monohexosyldihydroceramides), sphinganine, and sphinganine 1-phosphate. To test the hypothesis that elevation of sphinganine participates in the cytotoxicity of 4-HPR, cells were treated with the sphingosine kinase inhibitor d-erythro-N,N-dimethylsphingosine (DMS), with and without 4-HPR. After 24 h, the 4-HPR/DMS combination caused a 9-fold increase in sphinganine that was sustained through +48 hours, decreased sphinganine 1-phosphate, and increased cytotoxicity. Increased dihydrosphingolipids and sphinganine were also found in HL-60 leukemia cells and HT-29 cells treated with 4-HPR. The 4-HPR/DMS combination elicited increased apoptosis in all three cell lines. We propose that a mechanism of 4-HPR-induced cytotoxicity involves increases in dihydrosphingolipids, and that the synergy between 4-HPR and DMS is associated with large increases in cellular sphinganine. These studies suggest that enhanced clinical efficacy of 4-HPR may be realized through regimens containing agents that modulate sphingoid base metabolism.

Keyword: colon cancer

Alcohol Consumption-Related Metabolites in Relation to Colorectal and Adenoma: Two Case-Control Studies Using Serum Biomarkers.

Alcohol is a known carcinogen that may be associated with colorectal . However, most epidemiologic studies assess alcoholic beverage consumption using self-reported data, leading to potential exposure misclassification. Biomarkers of alcohol consumption may provide an alternative, complementary approach that reduces misclassification and incorporates individual differences in alcohol metabolism. Therefore, we evaluated the relationship between previously identified alcohol consumption-related metabolites and colorectal and adenoma using serum metabolomics data from two studies. Data on colorectal were obtained from a nested case-control study of 502 US adults (252 cases, 250 controls) within the Prostate, Lung, Colorectal, and Ovarian Screening Trial. Data on colorectal adenoma were obtained from a case-control study of 197 US adults (120 cases, 77 controls) from the Navy Adenoma Study. Unconditional multivariable logistic regression models were fit to calculate odds ratios (OR) and 95% confidence intervals (CI) for eight alcohol consumption-related metabolites identified in a previous analysis: ethyl glucuronide; 4-androstene-3beta,17beta-diol disulfate 1; 5-alpha-androstan-3beta,17beta-diol disulfate; 16-hydroxypalmitate; bilirubin (E,Z or Z,E); cyclo (-leu-pro); dihomo-linoleate (20:2n6); and palmitoleate (16:1n7). We found no clear association between these alcohol consumption-related metabolites and either endpoint. However, we did observe an inverse association between cyclo (-leu-pro) and colorectal adenoma that was only observed in the highest metabolite quantile (OR 4th vs. 1st Quantile = 0.30, 95% CI: 0.12-0.78; P-trend = 0.047), but no association for colorectal . In conclusion, there were no adverse associations between alcohol consumption-related metabolites and colorectal or adenoma.

Keyword: colon cancer

Role of acylethanolamides in the gastrointestinal tract with special reference to food intake and energy balance.

Acylethanolamides (AEs) are a group of lipids occurring in both plants and animals. The best-studied AEs are the endocannabinoid anandamide (AEA), the anti-inflammatory compound palmitoylethanolamide (PEA), and the potent anorexigenic molecule oleoylethanolamide (OEA). AEs are biosynthesized in the gastrointestinal tract, and their levels may change in response to noxious stimuli, food deprivation or diet-induced obesity. The biological actions of AEs within the gut are not limited to the modulation of food intake and energy balance. For example, AEs exert potential beneficial effects in the regulation of intestinal motility, secretion, inflammation and cellular proliferation. Molecular targets of AEs, which have been identified in the gastrointestinal tract, include cannabinoid CB(1) and CB(2) receptors (activated by AEA), transient receptor potential vanilloid type 1 (TRPV1, activated by AEA and OEA), the nuclear receptor peroxisome proliferators-activated receptor-alpha (PPAR-alpha, activated by OEA and, to a less extent, by PEA), and the orphan G-coupled receptors GPR119 (activated by OEA) and GPR55 (activated by PEA and, with lower potency, by AEA and OEA). Modulation of AE levels in the gut may provide new pharmacological strategies not only for the treatment of feeding disorders but also for the prevention or cure of widespread intestinal diseases such as inflammatory bowel disease and .

Keyword: colon cancer

Injury induced by fatty acids or bile in isolated human colonocytes prevented by calcium.

Measurement of the modulation of the growth fraction of isolated normal colonocytes from adult subjects in primary monolayer culture was used as a sensitive quantitative assay to evaluate toxic effects of several endogenous compounds found within the . This assay was used to study the role of CaCl2 in blocking cell injury. When added simultaneously with the injurious agent, 5-10 mM CaCl2 blocked the toxicity of physiological concentrations of deoxycholic , oleic , and linoleic .

Keyword: colon cancer

n-6 and n-3 polyunsaturated fatty acids differentially modulate oncogenic Ras activation in colonocytes.

Ras proteins are critical regulators of cell function, including growth, differentiation, and apoptosis, with membrane localization of the protein being a prerequisite for malignant transformation. We have recently demonstrated that feeding fish oil, compared with corn oil, decreases Ras membrane localization and reduces tumor formation in rats injected with a carcinogen. Because the biological activity of Ras is regulated by posttranslational lipid attachment and its interaction with stimulatory lipids, we investigated whether docosahexaenoic (DHA), found in fish oil, compared with linoleic (LA), found in corn oil, alters Ras posttranslational processing, activation, and effector protein function in young adult mouse cells overexpressing H-ras (YAMC-ras). We show here that the major n-3 polyunsaturated fatty (PUFA) constituent of fish oil, DHA, compared with LA (an n-6 PUFA), reduces Ras localization to the plasma membrane without affecting posttranslational lipidation and lowers GTP binding and downstream p42/44(ERK)-dependent signaling. In view of the central role of oncogenic Ras in the development of , the finding that n-3 and n-6 PUFA differentially modulate Ras activation may partly explain why dietary fish oil protects against development.

Keyword: colon cancer

Dietary soy sphingolipids suppress tumorigenesis and gene expression in 1,2-dimethylhydrazine-treated CF1 mice and ApcMin/+ mice.

Dietary supplementation with milk sphingolipids inhibits tumorigenesis in CF1 mice treated with a carcinogen [1,2-dimethylhydrazine (DMH)] and in multiple intestinal neoplasia (Min) mice, which develop intestinal tumors spontaneously. Plant sphingolipids differ structurally from those of mammals [soy glucosylceramide (GlcCer) consists predominantly of a 4,8-sphingadiene backbone and alpha-hydroxy-], which might affect their bioactivity. Soy GlcCer was added to the AIN-76A diet (which contains <0.005% sphingolipid) to investigate whether it would also suppress tumorigenesis in these mouse models. Soy GlcCer reduced cell proliferation in the upper half of the crypts in mice treated with DMH by 50 and 56% (P < 0.05) at 0.025 and 0.1% of the diet (wt/wt), respectively, and reduced the number of aberrant crypt foci (an early marker of carcinogenesis) by 38 and 52% (P < 0.05). Min mice fed diets containing 0.025 and 0.1% (wt/wt) soy GlcCer developed 22 and 37% fewer adenomas (P < 0.05), respectively. The effects of dietary sphingolipids on gene expression in the intestinal mucosal cells of Min mice were analyzed using Affymetrix GeneChip microarrays. Soy GlcCer affected the expression of 96 genes by > or = 2-fold in a dose-dependent manner, increasing 32 and decreasing 64. Decreases in the mRNA expression of two transcription factors associated with , hypoxia-induced factor 1 alpha (HIF1 alpha) and transcription factor 4 (TCF4), were confirmed by quantitative RT-PCR. In conclusion, soy GlcCer suppressed tumorigenesis in two mouse models; hence, plant sphingolipids warrant further investigation as inhibitors of . Because soy contains relatively high amounts of GlcCer, sphingolipids may partially account for the anticancer benefits attributed to soy-based foods.

Keyword: colon cancer

Tissue-Specific Distribution of Sphingomyelin Species in Pork Chop Revealed by Matrix-Assisted Laser Desorption/Ionization-Imaging Mass Spectrometry.

Sphingomyelin (SM) species are major sphingolipids in pork meat that affect quality parameters, such as health benefits due to their protective properties against chronic diseases; however, their spatial distribution remains unclear. We used matrix-assisted laser desorption/ionization (MALDI)-imaging mass spectrometry (IMS) to investigate the distribution and composition of SM species in pork chop consisting of longissimus thoracis et lumborum muscle (loin), intermuscular fat tissue, transparent tissue, and spinalis muscle. Four SM species were identified by liquid chromatography-electrospray ionization-tandem MS (MS/MS) and MALDI-MS/MS and visualized using MALDI-IMS. SM species containing stearic were predominantly distributed in the loin and spinalis muscle, whereas SM species containing , lignoceric, and nervonic acids were predominantly distributed in transparent tissue. These results indicated that the distribution of SM species differed among the pork tissues, depending on the tissue-specific fatty composition. The total amount including all identified SM species was higher in the loin than in spinalis muscle. Pork is reportedly associated with increased risk for chronic diseases due to the high amount of heme iron. From the observation of color, the amount of heme iron was lower in loin than in spinalis muscle. Thus, the degree of risk for chronic diseases might be lower in the loin than in spinalis muscle. This is the first report on the tissue-specific distribution of SM species in meat at a microscopic resolution using IMS. MALDI-IMS analysis may be useful in assessing the association between SM species and quality parameters of pork meat. PRACTICAL APPLICATION: Sphingomyelin (SM) species are major sphingolipids in pork meat. SM species affect quality parameters such as health benefits due to their protective properties against and atherosclerosis. Matrix-assisted laser desorption/ionization-imaging mass spectrometry analysis combined with liquid chromatography-electrospray ionization-tandem mass spectrometry is a suitable method to directly investigate the distribution and composition of SM species at microscopic level among different tissues of pork meat. Therefore, this method is useful to assess the SM species-induced health effect of different tissues of pork meat.© 2019 Institute of Food Technologists®.

Keyword: colon cancer

A bitter melon extract inhibits the P-glycoprotein activity in intestinal Caco-2 cells: monoglyceride as an active compound.

P-glycoprotein (P-gp) is a 170 kDa membrane protein that belongs to the ATP-binding cassette (ABC) transporter superfamily. In normal tissues, P-gp functions as an ATP-dependent efflux pump that excretes highly hydrophobic xenobiotic compounds, playing an important role in protecting the cells/tissues from xenobiotics. In the present study, chemical substances that could directly modulate the intestinal P-gp activity were searched in vegetables and fruits. By using human intestinal epithelial Caco-2 cells as a model of the small intestinal cells, we observed that a bitter melon fraction extracted from 40% methanol showed the greatest increase of the rhodamine-123 accumulation by Caco-2 cells. Inhibitory compounds in the bitter melon fraction were then isolated by HPLC using Pegasil C4 and Pegasil ODS columns. The HPLC fraction having the highest activity was analyzed by (1)H-NMR and FAB-MS, and the active compound was identified as 1-monopalmitin. It is interesting that certain types of monoglyceride might be involved in the drug bioavailability by specifically inhibiting the efflux mediated by P-gp.

Keyword: colon cancer

Acetyl-CoA carboxylase-alpha inhibitor TOFA induces human cell apoptosis.

Acetyl-CoA carboxylase-alpha (ACCA) is a rate-limiting enzyme in long chain fatty synthesis, playing a critical role in cellular energy storage and lipid synthesis. ACCA is upregulated in multiple types of human cancers and small interfering RNA-mediated ACCA silencing in human breast and prostate cells results in oxidative stress and apoptosis. This study reports for the first time that TOFA (5-tetradecyloxy-2-furoic ), an allosteric inhibitor of ACCA, is cytotoxic to lung cells NCI-H460 and carcinoma cells HCT-8 and HCT-15, with an IC(50) at approximately 5.0, 5.0, and 4.5 microg/ml, respectively. TOFA at 1.0-20.0 microg/ml effectively blocked fatty synthesis and induced cell death in a dose-dependent manner. The cell death was characterized with PARP cleavage, DNA fragmentation, and annexin-V staining, all of which are the features of the apoptosis. Supplementing simultaneously the cells with acids (100 microM), the end-products of the fatty synthesis pathway, prevented the apoptosis induced by TOFA. Taken together, these data suggest that TOFA is a potent cytotoxic agent to lung and cells, inducing apoptosis through disturbing their fatty synthesis.

Keyword: colon cancer

Identification of Palmitoylated Transitional Endoplasmic Reticulum ATPase by Proteomic Technique and Pan Antipalmitoylation Antibody.

Protein palmitoylation plays a significant role in a wide range of biological processes such as cell signal transduction, metabolism, apoptosis, and carcinogenesis. For high-throughput analysis of protein palmitoylation, approaches based on the acyl-biotin exchange or metabolic labeling of azide/alkynyl-palmitate analogs are commonly used. No palmitoylation antibody has been reported. Here, the palmitoylated proteome of human cell lines SW480 was analyzed via a TS-6B-based method. In total, 151 putative palmitoylated sites on 92 proteins, including 100 novel sites, were identified. Except for 3 known palmitoylated transmembrane proteins, ATP1A1, ZDHHC5, and PLP2, some important proteins including kinases, ion channels, receptors, and cytoskeletal proteins were also identified, such as CLIC1, PGK1, PPIA, FKBP4, exportin-2, etc. More importantly, the pan antipalmitoylation antibody was developed and verified for the first time. Our homemade pan antipalmitoylation antiserum could differentiate well protein palmitoylation from mouse brain membrane fraction and SW480 cells, which affords a new technique for analyzing protein palmitoylation by detecting the moiety directly. Furthermore, the candidate protein transitional endoplasmic reticulum ATPase (VCP) identified in SW480 cells was validated to be palmitoylated by Western blotting with anti-VCP antibody and the homemade pan antipalmitoylation antibody.

Keyword: colon cancer

Effects of specific fatty acids on cell transformation induced by an activated c-H-ras oncogene.

An increase in dietary lipid has been associated with an increase in the development of certain forms of , notably breast and , both in experimental animal studies and in human epidemiology studies. The underlying mechanisms are not, however, known with certainty. In the present studies we have examined whether certain specific fatty acids (FA) might act by enhancing the role of an activated oncogene in a model cell culture system. We found that when the rat fibroblast cell line Rat 6 was transfected with an activated human c-H-ras oncogene and the cells subsequently grown in medium supplemented with myristic , or stearic (20-80 microM) there was a marked enhancement of the number of transformed foci obtained. On the other hand arachidonic had a marked inhibitory effect in this transformation assay. However, this inhibitory effect can be partially reversed by indomethacin, an inhibitor of cyclo-oxygenase, at dose response manner. Control studies indicated that these results were not simply due to the effects of the FAs on growth of the Rat 6 cells or the process of transfection per se. Lipid analyses of cells grown in the presence of stearic indicated that the added FA was extensively incorporated into the major lipid classes of the cell and produced transient changes in lipid composition. This simple cell culture system may be useful for elucidating the mechanisms by which various dietary lipids and nutritional factors influence the carcinogenic process.

Keyword: colon cancer

Effects of fatty acids on liver metastasis of ACL-15 rat cells.

The effects of eicosapentaenoic [EPA; n-3 polyunsaturated fatty (PUFA)], linoleic (LA; n-6 PUFA), and (PA; saturated fatty ) on 1,2-dimethylhydrazine-induced F344 rat carcinoma cells (ACL-15) were investigated in vivo and in vitro. The number and size of liver metastatic foci via a superior mesenteric vein injection of ACL-15 cells in F344 rats were significantly inhibited in the EPA-treated group compared with the LA-treated group (p < 0.01); the PA-treated animals and those fed commercial rodent chow (standard diet) demonstrated intermediate values. In a dot immunoblotting assay, vascular cell adhesion molecule 1 expression on ACL-15 cells was downregulated by EPA-ethyl ester treatment and upregulated by LA-ethyl ester treatment compared with the untreated control cells, whereas the expression of matrix metalloproteinase 1 and 2 was not influenced by the fatty ethyl esters. In a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, EPA-ethyl ester suppressed ACL-15 cell growth in a schedule-dependent manner, and LA-ethyl ester showed schedule-dependent stimulation. In contrast, PA demonstrated no regulatory effect on cell growth at lower concentrations (< or = 5 mg/ml) but concentration-dependent inhibition at higher concentrations. According to our in vivo cell kinetic study, the difference in tumor growth at the metastatic site was due to different tumor cell proliferation rates; the cell loss rate was not altered. Therefore, the inhibitory effect of liver metastasis on ACL-15 cells by EPA can be explained by a decreased ability of tumor cell adhesion to the capillary bed (low expression of vascular cell adhesion molecule 1) and a lower potential of tumor cell proliferation (low mitotic rate) at the secondary site.

Keyword: colon cancer

Further insights into chemical characterization through GC-MS and evaluation for anticancer potential of Dracaena draco leaf and fruit extracts.

The present study reports for the first time the amino and fatty compositions and the antitumoral activity of aqueous extracts obtained from Dracaena draco L. leaf and fruit. Metabolite profiles were determined by gas chromatography-ion trap-mass spectrometry (GC-IT-MS), with several amino acids, , linolenic and stearic being identified in the leaf extract, and only proline, oleic and stearic in the fruit extract. The in vitro antiproliferative activities of the extracts were tested against human (Caco-2), kidney (A-498), and liver (HepG2) cell lines. In addition, primary cultures of normal and cancerous renal cells derived from kidney patients were treated with D. draco extracts (0-400 μg/mL). Antiproliferative and cytotoxic effects were determined by the MTT assay. D. draco extracts inhibited proliferation of human and renal tumor cells in vitro, whereas no or weak effect was observed in HepG2 cells. Compared to the fruit extract, D. draco leaf extract exhibited stronger antiproliferative activity against all cells. Our results indicate that D. draco, particularly the leaf, may be useful as a chemopreventive and/or chemotherapeutic agent for and kidney cancers.Copyright © 2012 Elsevier Ltd. All rights reserved.

Keyword: colon cancer

Fatty acylation of heparan sulfate proteoglycan from human carcinoma cells.

A number of transmembrane proteins have been recently reported to be modified by the covalent addition of saturated fatty acids which may contribute to membrane targeting and specific protein-lipid interactions. Such modifications have not been reported in cell-associated heparan sulfate proteoglycans, although these macromolecules are known to be hydrophobic. Here, we report that a cell surface heparan sulfate proteoglycan is acylated with both myristate and palmitate, two long-chain saturated fatty acids. When carcinoma cells were labeled with [3H]myristic , a significant proportion of the label was shown to be specifically incorporated into the protein core of the proteoglycan. Characterization of fatty acyl moiety in the purified proteoglycan by reverse-phase high pressure liquid chromatography revealed that approximately 60% of the covalently bound fatty acids was myristate. We further show that this relatively rare 14-carbon fatty was bound to the protein core via a hydroxylamine- and alkali-resistant amide bond. The remaining 40% was the more common 16-carbon palmitate, which was bound via a hydroxylamine- and alkali-sensitive thioester bond. Palmitate appeared to be added post-translationally and derived in part from intracellular elongation of myristate, a process that occurred within the first two hours and was insensitive to inhibition of protein synthesis. Acylation of heparan sulfate proteoglycan represents a novel modification of this gene product and could play a role in a number of biological functions including specific interactions with membrane receptors and ligand stabilization.

Keyword: colon cancer

Increased carnitine-dependent fatty uptake into mitochondria of human cells induces apoptosis.

Carnitine-dependent fatty import into mitochondria and beta-oxidation seem to be impaired in tumor cells. In the present study we show that a supply of palmitoylcarnitine together with L-carnitine potently induces apoptosis in HT-29 human cells as a consequence of accelerated fatty oxidation. Caspase-3-like activities, measured by the cleavage rate of a fluorogenic tetrapeptide substrate and nuclear fragmentation determined after DNA labeling in fixed cells by fluorescence microscopy, served as indicators of apoptosis. Neither L-carnitine nor palmitoylcarnitine alone were able to increase caspase-3-like activities and DNA fragmentation, but when provided together, apoptosis occurred. That exogenous carnitine was indeed able to enhance fatty uptake into mitochondria was demonstrated by an increased influx of a fluorescent analog. Enhanced fatty availability in mitochondria led to an increased generation of O*2-, as detected by a O*2- -sensitive fluorogenic dye, indicating oxidation of delivered substrates. Benzoquinone, an O*2- scavenger, blocked O*2- generation and prevented apoptosis as initiated by the combination of palmitoylcarnitine and carnitine. The lack of effect of the ceramide synthesis inhibitor fumonisin on palmitoylcarnitine/carnitine-induced apoptosis further supports the notion that apoptotic cell death is specifically due to fatty oxidation. In contrast to HT-29 cells, nontransformed human colonocytes did not respond to exogenous palmitoylcarnitine/carnitine and no apoptosis was observed. In conclusion, our studies provide evidence that a limited mitochondrial fatty import in human cells prevents high rates of mitochondrial O*2- production and protects cells from apoptosis that can be overcome by an exogenous carnitine supply.

Keyword: colon cancer

Hexadecanoic and neuraminic incorporations in two rat carcinoma cell lipids: selective influence of 1-O-octadecyl 2-O-methyl-3-phosphocholine on glycerolipid and ganglioside biosynthesis.

[3H] hexadecanoic and N-acetyl [14C] neuraminic acids were incorporated in glycerolipids or gangliosides of 2 rat carcinoma cell lines, having (PRO cells), or not (REG cells) invasive capacities when inoculated in syngeneic BD IX rats. The cells were cultured (48 h) in presence of 1-0-octadecyl-2-0-methyl-3-phosphocholine (ET 18-0-CH3) 20 or 40 microM, which, on transformed cells, inhibits the cell growth, modifies the glycerolipid biosynthesis, and activates the sialyltransferases. ET 18-0-CH3 20 microM activated, in PRO and in REG cells the incorporation of [3H] hexadecanoate in monosialogangliosides (1.45 fold compared to controls), but not in disialogangliosides and the distribution of this fatty between monosialo- (82%) and disialogangliosides (18%) was unchanged with controls. After [14C] neuraminic labelings, and for control experiments, the total radioactivities in gangliosides, in PRO cells, were twice higher than in REG cells, a difference which, probably, reflects the ganglioside content. ET 18-0-CH3 20 microM did not increase the incorporation of the [14C] neuraminic in PRO and in REG cells, and did not change its distribution between monosialo (70-80% for controls and experiments with ET 18-0-CH3) and disialogangliosides (20-30%). Similar results were obtained with ET 18-0-CH3 40 microM for the distribution of [14C] neuraminic in monosialo- and disialogangliosides. Whatever the precursor, the trisialogangliosides were never radiolabeled. Analysis of the [3H] glycerolipids (the main radiolabeled lipid classes in controls were: phosphatidylcholines, triglycerides, sphingomyelins and phosphatidyl-inositols) revealed that ET 18-0-CH3, compared to controls, did not activate the incorporation of [3H] hexadecanoate in total glycerolipids (PRO or REG cells). It activated (3 fold) its incorporation in triglyerides, inhibited it (0.5-0.6 fold) in phosphatidylcholines, sphingomyelins and phosphatidyl-inoditols and all these most noticeable differences were observed in PRO and in REG cells. These findings reflect the impossibility of ET 18-0-CH3 to activate the sialyltransferases during the ganglioside biosynthesis in carcinoma cells, while it modified ceramide, glycerophospholipid and neutral glycerolipid biosynthesis.

Keyword: colon cancer

PPARγ promotes diabetes-associated centrosome amplification via increasing the expression of SKA1 directly at the transcriptional level.

We have\xa0recently published that type 2 diabetes can induce cell centrosome amplification due to the action of high glucose, , and insulin, and ROCK1 and 14-3-3σ are signal mediators. In this study, we further investigated the molecular mechanisms of the centrosome amplification in HCT116 cells. Treatment of the cells with high glucose, , and insulin increased the expression of peroxisome proliferator-activated receptor γ (PPARγ) as well as the spindle and kinetochore associated protein 1 (SKA1), knockdown of each of which resulted in the inhibition of the treatment-triggered centrosome amplification. Knockdown of PPARγ inhibited the treatment-evoked increase in the SKA1 level, whereas knockdown of SKA1 did not modify the treatment-increased PPARγ level. We found a predicted binding site for PPARγ in the promoter region of the SKA1 gene from the JASPAR database. Experimental results showed that the treatment increased the messenger RNA level of SKA1, which could be inhibited by PPARγ chemical inhibitor or small interfering RNA. Moreover, we were able to show that PPARγ could bind to the binding site in the SKA1 gene promoter, which was increased by the experimental treatment. In conclusion, it is suggested that the pathophysiological factors in type 2 diabetes, high glucose, , and insulin, induce the cell centrosome amplification through the PPARγ-SKA1 pathway, in which PPARγ increases the expression of SKA1 via directly enhancing the SKA1 gene transcription.© 2019 Wiley Periodicals, Inc.

Keyword: colon cancer

Neutral glycolipid composition in human .

Neutral glycolipid composition of human gastric and was studied in comparison to healthy mucosa. The following compounds were identified: galactosylceramide, glucosylceramide, trihexosylceramide, globoside, small amounts of neotetraosylceramide and fucoglycolipids. The latter comprised both 3-0- and 4-0-substituted residues of N-acetylglucosamine. In tissue the contents of lactosylcermide and of fucoglycolipids were increased. Fatty compositions of neutral glycolipids derived from both and control samples were similar. The most abundant fatty acids were and nervonic acids.

Keyword: colon cancer

Dietary and biomarker estimates of fatty acids and risk of colorectal .

The associations between intake of or circulating fatty acids and risk of colorectal (CRC) are unclear. We examined prospectively the associations between dietary or biomarker fatty acids and CRC. For 41,514 men and women, aged 40-69 years, baseline (1990-94) dietary intakes of fatty acids were estimated using a food frequency questionnaire and plasma phospholipid (PPL) fatty acids were measured for 4,205 participants including 395 CRC cases, according to a case-cohort design. Hazard ratios were computed using Cox regression adjusting for education, alcohol intake, smoking status, physical activity and total energy intake; and stratified for gender, ethnicity and family history of , with age as the time scale. We assessed the heterogeneity of associations with and rectal cancers. PPL saturated fatty acids (SFAs) were positively associated with CRC risk, while total n-3 polyunsaturated fatty acids (PUFA) and long chain marine n-3 PUFAs showed inverse associations, significant only for 22:5 n-3. No significant associations were observed for dietary fatty intakes but positive associations with CRC of borderline significance were seen for both dietary and PPL linoleic . Positive associations with dietary (16:0), MUFAs and n-6 PUFAs were seen for rectal but not cancers. PPL 22:6 n-3 was inversely associated with rectal . Limiting intakes of SFAs and MUFAs could be assisted by following existing guidelines to limit red and processed meats which are important sources in the Australian diet. Our observations regarding linoleic should be examined further.© 2015 UICC.

Keyword: colon cancer

Effect of dietary tetradecylthioacetic on growth studied by dynamic contrast enhanced MRI.

To evaluate the effects of diets supplemented with the modified fatty tetradecylthioacetic (TTA) and fish protein hydrolysate (FPH) on tumor growth of the human cell line SW620, and to investigate the properties of tumor vasculature by dynamic contrast-enhanced MRI in a human tumor xenograft.SW620 cells were grown in vitro in presence of TTA and and proliferation was measured by thymidine incorporation. The xenograft study in mice was performed with four distinct diets: (a) control diet; (b) diet with TTA; (c) diet with TTA and FPH; and (d) diet with FPH. SW620 cells were injected subcutaneously, and dynamic contrast enhanced (DCE) MRI was performed on a Bruker BioSpec 7T system. The data was analyzed by two-compartment modeling of the contrast enhancement, initial area under the curve (IAUC) and by use of relative signal intensity (RSI) distributions.The in vitro cell studies revealed that TTA reduced tumor cell proliferation as a function of both dose and time. The in vivo tumor growth was significantly reduced for the two groups fed TTA, as compared to the control group. The mean 10(th) percentile RSI, v(e) and IAUC for the TTA group were significant higher than for the control group.This study confirms the growth inhibitory effects of TTA, both in vitro and in vivo, in a model. The analysis of DCE-MRI data showed that TTA influences the vascular properties of the tumor in addition to the growth.

Keyword: colon cancer

Fatty esterification during differentiation of the human intestinal cell line Caco-2.

The Caco-2 human intestinal cell line was used to examine fatty esterification during development of the enterocytic phenotype. Acyl-CoA synthetase activity increased approximately 40%, and the incorporation of into triacylglycerol relative to phosphatidylcholine increased nearly 2-fold during Caco-2 differentiation. A rate-limiting enzyme activity in the glycerol 3-phosphate pathway of triacylglycerol synthesis, glycerol-3-phosphate acyltransferase, was at levels comparable with rat jejunum and remained unchanged during differentiation. In contrast, the activity of monoacylglycerol acyltransferase, which is unique to the monoacylglycerol pathway of triacylglycerol synthesis, was present at < 7% of the levels in rat jejunum. Further analysis of the glycerol 3-phosphate pathway showed that the rate-limiting enzyme activities for diacylglycerol conversion to triacylglycerol, diacylglycerol acyltransferase, and phosphatidylcholine, CTP:phosphocholine cytidylyltransferase, increased 2-3-fold and decreased approximately 40%, respectively, during Caco-2 differentiation. In addition, a 2-fold increase in cellular diacylglycerol mass was observed during enterocytic conversion. These data indicate that fatty esterification to triacylglycerol in Caco-2 cells occurs primarily via the glycerol 3-phosphate pathway. Furthermore, the differentiation-dependent increase in fatty esterification to triacylglycerol relative to phosphatidylcholine appears to result from increased utilization of diacylglycerol to synthesize triacylglycerol and a concomitant decrease in diacylglycerol utilization for phosphatidylcholine synthesis.

Keyword: colon cancer

PCNA and JNK1-Stat3 pathways respectively promotes and inhibits diabetes-associated centrosome amplification by targeting at the ROCK1/14-3-3σ complex in human HCT116 cells.

We have recently reported that type 2 diabetes promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho-associated coiled-coil containing protein kinase 1 (ROCK1)/14-3-3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14-3-3σ. Individual knockdown of PCNA, NPM, or 14-3-3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment-increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and also increased the expressions of c-Jun N-terminal kinase-1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment-increased expression of 14-3-3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14-3-3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment-increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1-Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14-3-3σ complex, and NPM serves as an independent signal for the centrosome amplification.© 2018 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

Keyword: colon cancer

and linoleic metabolism in Caco-2 cells: different triglyceride synthesis and lipoprotein secretion.

Polarized monolayers of intestinal Caco-2 cells were used to study the effects of saturated (16:0) and polyunsaturated linoleic (18:2) on triglyceride synthesis and lipoprotein secretion. Monolayers were incubated for 24 h, at the apical or lumenal side, with (16:0) or linoleic (18:2) in physiological concentrations. Incubation with 1.0 mM 16:0 or 18:2 resulted in differences in the composition and amount of secreted lipoproteins. Radiolabeled lipids in the lipoproteins secreted during incubation with 18:2 were found in the chylomicron/VLDL (very low density lipoprotein) density whereas with 16:0 the secreted lipoproteins were in the intermediate density/low density lipoprotein (IDL/LDL) density range. More triglyceride was secreted into the (basolateral) medium during incubation with 1.0 mM 18:2 (41 +/- 12% of total triglyceride synthesized) than with 1.0 mM 16:0 (18 +/- 3% of total). The biochemical findings correlate with conspicuous morphological changes in the cells in the presence of 16:0, but not 18:2. Increasing concentrations of 16:0 (0.1-1.0 mM) caused gradual accumulation of intracellular membrane. Microvilli became strongly reduced in number. With 1.0 mM we found an increased incorporation of [1-14C] into phosphatidic (14.8% of total incorporation into phospholipid with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. 0.5% with 18:2) and the amount of intracellular phospholipid doubled. The morphological changes were completely reversed after 24 h with 1.0 mM 18:2. We conclude from our results that, compared to 18:2, 16:0 is not efficiently incorporated into triglycerides. 16:0 is incorporated into cellular phospholipids in a greater proportion than 18:2, causing accumulation of intracellular phospholipid and the precursors phosphatidic and diacylglycerol. Different processing of 18:2 and 16:0 by Caco-2 cells resulted in profound differences in triglyceride synthesis and lipoprotein composition and secretion.

Keyword: colon cancer

Biochemical epidemiology of : effect of types of dietary fiber on diacylglycerols in women.

In view of the potential significance of dietary fat and fiber in and the possible indirect involvement of diacylglycerols (DAGs) in the pathogenesis of , the effect of types of dietary fiber on fecal DAG in premenopausal women was investigated.Forty-eight women consuming a typical western diet provided two 24-hour stool specimens and two sets of preintervention 4-day food records. They were randomly assigned to one of the fiber groups, namely, a wheat, oat, or corn bran supplement. They consumed their control diet plus 13-15 g of dietary fiber from each source for 8 weeks. At the end of the fiber period, each subject provided two 24-hour stool specimens and 4-day food records. Stool samples collected during the two periods were analyzed for total fat and DAG fatty acids.All sources of dietary fiber increased the amount of fecal fat excreted. Dietary wheat bran decreased the concentrations of total DAG and DAG containing lauric , myristic , , stearic , and linoleic , whereas oat bran increased the DAG composed of oleic and linoleic compared with the control diet. Corn bran decreased the DAG containing stearic .These results show that the modifying effect of dietary fiber on DAG depends on the type of fiber consumed.

Keyword: colon cancer

Modification of CaCo-2 cell membrane fatty composition by eicosapentaenoic and : effect on cholesterol metabolism.

Membrane fatty composition of CaCo-2 cells was modified by incubating the cells for 8 days in medium containing 100 microM eicosapentaenoic or . The effect of membrane fatty changes on cholesterol metabolism was then studied. Cells incubated with eicosapentaenoic had significant changes in membrane fatty composition with an accumulation of 20:5 and 22:5 and a reduction in monoenoic fatty acids compared to cells grown in . Intracellular cholesteryl esters could not be detected in CaCo-2 cells grown in the presence of the n-3 polyunsaturated fatty . In contrast, cells incubated with the saturated fatty contained 2 micrograms/mg protein of cholesteryl esters. Cells grown in eicosapentaenoic , however, accumulated significantly more triglycerides compared to cells modified with . The rate of oleic incorporation into triglycerides was significantly increased in cells incubated with eicosapentaenoic . CaCo-2 cells modified by eicosapentaenoic had lower rates of HMG-CoA reductase and ACAT activities compared to cells modified with . The incorporation of the two fatty acids into cellular lipids also differed. was predominantly incorporated into cellular triglycerides, whereas eicosapentaenoic was preferentially incorporated into phospholipids with 60% of it in the phosphatidylethanolamine fraction. The data indicate that membrane fatty composition is significantly altered by growing CaCo-2 cells in eicosapentaenoic . These modifications in membrane fatty saturation are accompanied by a decrease in the rates of cholesterol synthesis and cholesterol esterification.

Keyword: colon cancer

New flavan and alkyl alpha,beta-lactones from the stem bark of Horsfieldia superba.

In the present study phytochemical investigation of the methanol extract of the stem bark of Horsfieldia superba led to the isolation of twenty compounds (1-20), of which three (1-3) were new. However, compounds 2 and 3 were previously reported as synthetic alpha,beta-lactones. The compounds were characterized as (-)-3,4',7-trihydroxy-3'-methoxyflavan (1), (-)-5,6-dihydro-6-undecyl-2H-pyran-2-one (2), and (-)-5,6-dihydro-6-tridecyl-2H-pyran-2-one (3). Seventeen other known compounds were also isolated and identified as (-)-viridiflorol (4), hexacosanoic (5), beta-sitosterol (6), methyl 2,4-dihydroxy-6-methylbenzoate (methylorsellinate) (7), methyl 2,4-dihydroxy-3,6-dimethylbenzoate (8), (-)-4'-hydroxy-7-methoxyflavan (9), (-)-4',7-dihydroxyflavan (10), (-)-4',7-dihydroxy-3'-methoxyflavan (11), (+)-3,4',7-trihydroxyflavan (12), (-)-catechin (13), (-)-epicatechin (14), (-)-7-hydroxy-3',4'-methylenedioxyflavan (15), 2',3,4-trihydroxy-4'-methoxydihydrochalcone (16), 3',4',7-trihydroxyflavone (17), (+)-4'-hydroxy-7-methoxyflavanone (18), hexadecanoic () (19) and 3,4-dihydroxybenzoic (20). The structures of the compounds were fully characterized by various physical methods (melting point, optical rotation), spectral (UV, IR, ID and 2D NMR) and mass spectrometric techniques. In vitro assay of compounds 2 and 3 demonstrated moderate cytotoxic activities against human prostate (PC-3), (HCT-116) and breast (MCF-7) cells, while the chloroform and ethyl acetate fractions of H. superba were found to exhibit moderate AChE inhibitory activity (IC50 72 and 60 microg/mL).

Keyword: colon cancer

Altered Saturated and Monounsaturated Plasma Phospholipid Fatty Profiles in Adult Males with Adenomas.

Altered lipid metabolism and plasma fatty (FA) levels are associated with colorectal . Obesity and elevated waist circumference (WC) increase the likelihood of developing precancerous adenomas.Venous blood was collected from 126 males, ages 48 to 65 years, who received routine colonoscopies. Plasma phospholipid (PPL) FAs were isolated, derivatized, and then analyzed using gas chromatography. ORs and 95% confidence intervals were determined using polytomous logistic regression after adjusting for confounding factors [i.e., age, smoking, WC, and body mass index (BMI)].PPL (PA) was inversely correlated with the presence of adenomas (P = 0.01). For each unit increase in palmitoleic (OR, 3.75; P = 0.04) or elaidic (OR, 2.92; P = 0.04), an individual was more likely to have adenomas relative to no polyps. Higher enzyme activity estimates (EAE) of stearoyl-CoA desaturase-1 (SCD-1; P = 0.02) and elongation of very long chain fatty acids protein-6 (ELOVL-6; P = 0.03) were associated with an individual being approximately 1.5 times more likely to have an adenoma compared with no polyps.PPL FAs and EAEs, which have previously been associated with colorectal , are significantly different in those with adenomas when compared with those without polyps. PPL PA, elaidic , and SCD-1 and ELOVL-6 EAEs are associated with adenomas independent of BMI and WC.PPL PA, elaidic , and SCD-1 and ELOVL-6 EAEs are associated with adenomas even after adjusting for obesity-related risk factors and may function as novel biomarkers of early colorectal risk.©2015 American Association for Research.

Keyword: colon cancer

Non-proteolytic release of carcinoembryonic antigen from normal human epithelial cells cultured in collagen gel.

Recent studies have shown that, even with a minimal content of carcinoembryonic antigen (CEA), normal human epithelial cells express substantial amounts of CEA mRNA and mucosal fragments cultured in vitro produce CEA quite actively, indicating that CEA should no longer be considered to be of an oncofetal nature. To understand the basis of the usefulness of CEA as a tumor marker, we analyzed the release of CEA, a glycosyl-phosphatidylinositol (GPI)-anchored protein, from epithelial cells, by culturing isolated crypts in collagen gel. The crypts appeared to preserve their morphological and biochemical integrity in the gel for at least 16 hr, and released CEA spontaneously. Three forms of CEA--spontaneously released CEA, CEA liberated with phosphatidylinositol-specific phospholipase C (PI-PLC) and CEA in cell lysates--were indistinguishable on SDS-PAGE. This is in contrast to recombinant CEA spontaneously released from CHO transfectants, which showed a smaller molecular mass than that of PI-PLC-cleaved recombinant CEA. By phase separation using Triton X-114, CEA in the cell lysates of crypts was separated mostly into the detergent phase, while the spontaneously released and the PI-PLC-cleaved CEA were separated into the aqueous phase. When the cells were metabolically labeled with the precursors of the GPI-anchor, 3H-ethanolamine but not 3H- was found in the spontaneously released CEA. These findings suggest that, in contrast to the proteolysis-like release of the recombinant CEA from CHO cells, CEA in normal epithelial cells is released by a non-proteolytic cleavage, which probably occurs through the action of some endogenous phospholipase.

Keyword: colon cancer

Antitumor effect of -conjugated DsiRNA for in a mouse subcutaneous tumor model.

In this study, we synthesized Dicer-substrate siRNA conjugated with at the 5'-end of the sense strand (C16-DsiRNA), and examined its RNAi effect on β-catenin as a target gene in a cell line, HT29Luc, both in vitro and in vivo. We examined the in vitro RNAi effect in HT29Luc cells and found that C16-DsiRNA strongly inhibited expression of the β-catenin gene in comparison with non-modified DsiRNA. Also, high membrane permeability of C16-DsiRNA was exhibited, and it was confirmed that most of the C16-DsiRNA was localized in cytoplasm of HT29Luc cells. In regard to the in vivo RNAi effect, C16-DsiRNA complexed with Invivofectamine targeting the β-catenin gene was locally administered to a subcutaneous tumor formed by implantation of HT29Luc cells into the subcutis of nude mice; we evaluated the effect by measuring the bioluminescence increase, which reflects tumor growth, using an in vivo imaging system. As a result, C16-DsiRNA strongly inhibited the growth of tumors formed in subcutis of nude mice compared with non-modified DsiRNA, and this in vivo RNAi effect lasted up to 15\xa0days. Our results suggest that C16-DsiRNA should be vigorously pursued as a novel nucleic medicine for clinical treatment of .© 2018 John Wiley & Sons A/S.

Keyword: colon cancer

Urokinase secretion from human carcinomas induced by endogenous diglycerides.

tumor cells are more responsive to certain growth modulators in their local environment in vivo than are normal colonocytes. Examples of this class of compounds are the fecal diglycerides (DGs)(E. Friedman et al., Res., 49: 544-548, 1989), which may act as endogenous tumor promoters. At the concentration found in vivo, fecal DGs composed of oleic, myristic, and fatty acids induced mitogenesis of all classes of benign tumor cells and of half of the resected carcinomas tested in primary culture, but induced no detectable mitogenesis of normal colonocytes. tumor cells also exhibit selective responses to these endogenous modulators as measured by another biological parameter, secretion of urokinase from carcinomas than from normal colonocytes. Fecal DGs also induced a 13-fold increase in urokinase mRNA synthesis in carcinoma cells and induced secretion of active urokinase from each of five resected carcinomas. carcinomas, at both the primary site and metastatic to the liver, secreted the Mr 55,000 form of urokinase constitutively and secreted the same form upon treatment with fecal DGs. An increase in the steady-state level of urokinase secretion by saturated-chain DGs exhibited a strong dependency on the chain length of the fatty residues, those of 14 and 16 carbons having the greatest activity. Thus, fecal DGs composed of oleic, myristic, and residues induce two biological activities selectively in tumor cells, each of which would enhance tumor development. Selective mitogenesis would increase adenoma and carcinoma cell number relative to normal colonocyte number, and induction of the proteolytic enzyme urokinase would aid local invasion of the carcinoma within the bowel wall.

Keyword: colon cancer

Hexadecenoic Fatty Positional Isomers and De Novo PUFA Synthesis in Cells.

metabolism involves delta-9 and delta-6 desaturase enzymes forming palmitoleic (9-16:1; -7 series) and sapienic (6-16:1; -10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty (MUFA) isomers are under development. Furthermore, sapienic can bring to the de novo synthesis of the -10 polyunsaturated fatty (PUFA) sebaleic (5,8-18:2), but such transformations in cells are not known. The model of Caco-2 cell line was used to monitor sapienic supplementation (150 and 300 μM) and provide evidence of the formation of -10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and acids evidenced that lipid remodelling was influenced by the type of fatty and positional isomer, with an increase of 8-18:1, -10 PUFA and a decrease of saturated fats in case of sapienic . Cholesteryl esters were formed only in cases with sapienic . Sapienic was the less toxic among the tested fatty acids, showing the highest ECs and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic increases the distribution of fluid regions, probably connected with the formation of 8-18:1 and the -10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and research.

Keyword: colon cancer

Incorporation of bile of low concentration into model and biological membranes studied by 2H and 31P NMR.

We have analyzed the manner of incorporation of bile into lipid bilayers and resultant perturbation of the bilayer structure with lower bile /lipid ratios relevant to the physiological conditions (approximately 1 mM) by 2H and 31P NMR methods, as an aid to understanding the possible role as an endogenous tumor promoter in besides the primary physiological function of solubilizing lipids. On the basis of the 2H quadrupole splittings of [6,6,7,7,8-2H5]deoxycholate and [11,11,12,12-2H4]chenodeoxycholate in the presence of lamellar multibilayers of egg yolk lecithin, these bile acids were found to be incorporated in such a manner that the B-D rings lie parallel with the normal of the bilayers when the ratio of the bile to lipid is low (less than 0.11). When the ratio is increased, these bile molecules are not dispersed entirely in the bilayer but aggregate to form micelles with lipids. Further, we studied the resultant perturbation of the multibilayers of egg yolk lecithin analyzed by using the 2H quadrupole splitting of [18,18,18-2H3]stearic as a probe and by 31P chemical shift anisotropy. We found that the bilayer structure is retained even at the bile -to-lipid ratio of 0.25, although a small amount of an isotropic phase appeared such as small vesicles and micelles. The molecular ordering of fatty acyl chains was rather enhanced by the presence of 1 mM deoxycholate in erythrocyte ghosts as seen from the 2H quadrupole splitting of [16,16,16-2H3], although deoxycholate caused hemolysis in this condition. The former observation can be explained by the way the lipid-protein interaction is modified by deoxycholate located in the interface between the lipids and proteins.

Keyword: colon cancer

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Anti-Escherichia coli O157:H7 activity of free fatty acids under varying pH.

Following screening of 4 strains of Escherichia coli O157:H7 (E32511, E318N, H4420N, and R508N) for tolerance, strain H4420N was selected for further study into the influence of pH on bactericidal activity of 6 fatty acids (capric, lauric, , oleic, linoleic, and linolenic). Strain H4420N was cultured for 6 h in Luria-Bertani broth amended with individual fatty acids at 20 mmol/L, with pH adjusted to 7.0, 4.3, or 2.5. None of the fatty acids exhibited bactericidal activity at pH 7.0 (p >0.05). At pH 4.3, only capric, lauric, and linoleic acids reduced viability of H4420N (p < 0.05). At pH 2.5, oleic (C18:1) and linolenic (C18:3) acids had modest effects on H4420N viability, whereas capric (C10:0), lauric (C12:0), and linoleic (C18:2) acids resulted in a reduction > or =5 log10 colony-forming units (CFU)/mL (p < 0.05). Capric and lauric acids were examined further at pH 2.5 over a range of concentrations (0.15-20 mmol/L). After 10 min of exposure, 5 log10 CFU/mL reductions (p < 0.05) were achieved by lauric at 2.5 mmol/L and by capric at 0.31 mmol/L. stress increased the sensitivity of -tolerant E. coli O157:H7 strain H4420N to fatty acids. Including sources of these fatty acids in diets for cattle might impair the ability of this zoonotic pathogen to survive passage through the stomach, possibly reducing the potential for its in the lower gut.

Keyword: colonization

Metabolite profiling of mycorrhizal roots of Medicago truncatula.

Metabolite profiling of soluble primary and secondary metabolites, as well as cell wall-bound phenolic compounds from roots of barrel medic (Medicago truncatula) was carried out by GC-MS, HPLC and LC-MS. These analyses revealed a number of metabolic characteristics over 56 days of symbiotic interaction with the arbuscular mycorrhizal (AM) fungus Glomus intraradices, when compared to the controls, i.e. nonmycorrhizal roots supplied with low and high amounts of phosphate. During the most active stages of overall root mycorrhization, elevated levels of certain amino acids (Glu, Asp, Asn) were observed accompanied by increases in amounts of some fatty acids ( and oleic acids), indicating a mycorrhiza-specific activation of plastidial metabolism. In addition, some accumulating fungus-specific fatty acids (palmitvaccenic and vaccenic acids) were assigned that may be used as markers of fungal root . Stimulation of the biosynthesis of some constitutive isoflavonoids (daidzein, ononin and malonylononin) occurred, however, only at late stages of root mycorrhization. Increase of the levels of saponins correlated AM-independently with plant growth. Only in AM roots was the accumulation of apocarotenoids (cyclohexenone and mycorradicin derivatives) observed. The structures of the unknown cyclohexenone derivatives were identified by spectroscopic methods as glucosides of blumenol C and 13-hydroxyblumenol C and their corresponding malonyl conjugates. During mycorrhization, the levels of typical cell wall-bound phenolics (e.g. 4-hydroxybenzaldehyde, vanillin, ferulic ) did not change; however, high amounts of cell wall-bound tyrosol were exclusively detected in AM roots. Principal component analyses of nonpolar primary and secondary metabolites clearly separated AM roots from those of the controls, which was confirmed by an hierarchical cluster analysis. Circular networks of primary nonpolar metabolites showed stronger and more frequent correlations between metabolites in the mycorrhizal roots. The same trend, but to a lesser extent, was observed in nonmycorrhizal roots supplied with high amounts of phosphate. These results indicate a tighter control of primary metabolism in AM roots compared to control plants. Network correlation analyses revealed distinct clusters of amino acids and sugars/aliphatic acids with strong metabolic correlations among one another in all plants analyzed; however, mycorrhizal symbiosis reduced the cluster separation and enlarged the sugar cluster size. The amino clusters represent groups of metabolites with strong correlations among one another (cliques) that are differently composed in mycorrhizal and nonmycorrhizal roots. In conclusion, the present work shows for the first time that there are clear differences in development- and symbiosis-dependent primary and secondary metabolism of M. truncatula roots.

Keyword: colonization

Deficient production of hexadecenoic in the skin is associated in part with the vulnerability of atopic dermatitis patients to by Staphylococcus aureus.

As one of the major skin fatty acids, cis-6-hexadecenoic (C16:1Delta6) exhibits a specific antibacterial activity and might play a specific role in the defense mechanism against Staphylococcus aureus, in healthy subjects whereas S. aureus frequently colonizes the skin of patients with atopic dermatitis (AD).Fatty composition of sebum at the recovery level was analyzed by gas chromatography and S. aureus colonizing the skin was assessed by the \'cup-scrub\' method (9 patients and 10 healthy controls). To evaluate in vivo effect of C16:1Delta6 on , C16:1Delta6 was applied for 2 weeks on the upper arm skin of another group of AD patients (11 patients).Analysis of sebum lipids revealed that there is a significant lower free C16:1Delta6 content in nonlesional skin from AD patients compared with healthy controls. This lower content is also accompanied by a significantly lower level of C16:1Delta6 in the total fatty composition of sebum (analyzed following hydrolysis). The lower level of free C16:1Delta6 correlated significantly (R(2) = 0.41, p < 0.01) with the numbers of S. aureus colonizing nonlesional skin. Topical application of free C16:1Delta6 on the skin of AD patients for 2 weeks abolished the markedly increased bacterial count in 6 out of the 8 AD patients tested.Free C16:1Delta6 may be involved in the defense mechanism against S. aureus in healthy skin and this deficit triggers the susceptibility of the skin to by S. aureus in AD.Copyright 2005 S. Karger AG, Basel.

Keyword: colonization

Human antibody response to outer membrane protein G1a, a lipoprotein of Moraxella catarrhalis.

Moraxella catarrhalis is an important cause of respiratory infections in adults with chronic obstructive pulmonary disease (COPD) and of otitis media in children. Outer membrane protein (OMP) G1a is an approximately 29-kDa surface lipoprotein and is a potential vaccine candidate. The gene that encodes OMP G1a was expressed and purified using a novel plasmid vector. [(3)H] labeling demonstrated that both native and recombinant OMP G1a contain covalently bound . To assess the expression of OMP G1a during human infection, paired sera and sputum supernatants from adults with COPD followed prospectively were studied by enzyme-linked immunosorbent assays with recombinant lipidated OMP G1a to detect antibodies made specifically during carriage of M. catarrhalis. Overall, 23% of patients developed either a serum immunoglobulin G (IgG) response (9%) or sputum IgA response (21%) to OMP G1a, following 100 episodes of acquisition and clearance of M. catarrhalis. Patients developed antibody responses at similar rates following episodes of clinical exacerbation compared to asymptomatic . Serum IgG antibodies following natural infection were directed predominantly at OMP G1a epitopes that are not exposed on the bacterial surface. These data show that OMP G1a is expressed during infection of the human respiratory tract and is a target of systemic and mucosal antibodies. These observations indicate that OMP G1a, a highly conserved surface protein, should be evaluated further as a vaccine candidate.

Keyword: colonization

In vitro and in vivo exploration of from Synechococcus elongatus as an antibiofilm agent on the survival of Artemia franciscana against virulent vibrios.

Biofilm formation of Vibrio spp. has been demonstrated as a potentially important mechanism contributing antibiotic treatment failure in aquaculture. In the present study, the effect of (PA) identified from Synechococcus elongatus was assessed for the inhibition of quorum sensing (QS) regulated biofilm formation in aquatic bacterial pathogens. The biofilm inhibitory concentration (BIC) of PA against Vibrio spp. was found to be 100µgml. In this concentration, PA exhibited a significant inhibition in biofilm biomass of Vibrio harveyi MTCC 3438, V. parahaemolyticus ATCC 17802, V. vulnificus MTCC 1145 and V. alginolyticus ATCC 17749 without hindering their planktonic growth. Also, PA displayed gradual decrease in bioluminescence production of V. harveyi. The results of extracellular polymeric substances quantification, microbial adhesion to hydrocarbons and Fourier transform infrared spectroscopic (FT-IR) analyses suggested that PA positively interferes with the initial adhesion stages of biofilm formation. In addition, confocal and scanning electron microscopic analysis substantiates the antibiofilm efficacy of the PA. The transcriptomic analysis revealed the down-regulation of QS mediated response regulator genes expression in V. harveyi. Concomitantly, PA reduced the intestinal of vibrios in brine shrimp larvae and thereby attenuates the biofilm assemblage and its associated virulence. In vivo studies using brine shrimp larvae manifested the reduction in adherence and virulence, which prompts further investigation about the potential of PA for the treatment of vibriosis.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: colonization

Enterobacter sp. strain Fs-11 adapted to diverse ecological conditions and promoted sunflower achene yield, nutrient uptake, and oil contents.

Plant growth-promoting rhizobacteria are under extensive investigation to supplement the chemical fertilizers due to cost-effective and eco-friendly nature. However, their consistency in heterogeneous soil and diverse ecological settings is unclear. The current study presents in vitro and field evaluation of pre-characterized PGPR strain Enterobacter sp. Fs-11 (GenBank accession # GQ179978) in terms of its potential to enhance sunflower yield and oil contents under diverse environmental conditions. Under in vitro conditions, strain Fs-11 showed optimal growth at a range of temperature (15 to 40\xa0°C) and pH values (6.5 to 8.5). Extracellular and intracellular localizations of the strain Fs-11 in sunflower root cortical cells through transmission electron microscopy confirmed its epiphytic and endophytic patterns, respectively. In field experiments, conducted at three different agro-climatic locations, inoculation of strain Fs-11 at 50% reduced NP fertilizer resulted in a significant increase in growth, achene yield, nutrient uptake, and oil contents. Inoculation also responded significantly in terms of increase in mono- and polyunsaturated fatty acids (oleic and linoleic acids, respectively) without rising saturated fatty ( and stearic acids) contents. We concluded that Enterobacter sp. Fs-11 is a potential candidate for biofertilizer formulations to supplement chemical fertilizer requirements of sunflower crop under diverse climatic conditions.

Keyword: colonization

Hydrophobicity of cultured mammalian cells and some effects of bacterial phospholipases C.

Eucaryotic cell surface hydrophobicity was measured as a partition of between the cell surface and the surrounding buffer. The method was found to be independent of cell mass or amount of within a large interval. An estimation of cell stability could also be obtained. The effects of Ca and Mg ions on cell hydrophobicity and stability of mouse myeloma (SP2/O) cells and of Chinese hamster ovary (CHO) cells were determined. This system permits measurement of changes in cell hydrophobicity caused by various additives, e.g. ions, purified bacterial products, antibiotics or cytostatics. Studies were made on these eucaryotic cells treated with purified bacterial phospholipases C from S. aureus and C. perfringens. These enzymes were found to increase the eucaryotic cell membrane hydrophobicity. This finding might indicate that bacterial phospholipases C facilitate bacterial at the start of an infection.

Keyword: colonization

Bactericidal activity of the human skin fatty cis-6-hexadecanoic on Staphylococcus aureus.

Human skin fatty acids are a potent aspect of our innate defenses, giving surface protection against potentially invasive organisms. They provide an important parameter in determining the ecology of the skin microflora, and alterations can lead to increased by pathogens such as Staphylococcus aureus. Harnessing skin fatty acids may also give a new avenue of exploration in the generation of control measures against drug-resistant organisms. Despite their importance, the mechanism(s) whereby skin fatty acids kill bacteria has remained largely elusive. Here, we describe an analysis of the bactericidal effects of the major human skin fatty cis-6-hexadecenoic (C6H) on the human commensal and pathogen S. aureus. Several C6H concentration-dependent mechanisms were found. At high concentrations, C6H swiftly kills cells associated with a general loss of membrane integrity. However, C6H still kills at lower concentrations, acting through disruption of the proton motive force, an increase in membrane fluidity, and its effects on electron transfer. The design of analogues with altered bactericidal effects has begun to determine the structural constraints on activity and paves the way for the rational design of new antistaphylococcal agents.Copyright © 2014 Cartron et al.

Keyword: colonization

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula.

The peroxidation of polyunsaturated fatty acids, common to all eukaryotes, is mostly catalyzed by members of the lipoxygenase enzyme family of non-heme iron containing dioxygenases. Lipoxygenase products can be metabolized further in the oxylipin pathway by several groups of CYP74 enzymes. One prominent oxylipin is jasmonic (JA), a product of the 13-allene oxide synthase branch of the pathway and known as signaling substance that plays a role in vegetative and propagative plant development as well as in plant responses to wounding and pathogen attack. In barley roots, JA level increases upon by arbuscular mycorrhizal fungi. Apart from this first result regarding JA, no information is available on the relevance of lipidperoxide metabolism in arbuscular mycorrhizal symbiosis. Thus we analyzed fatty and lipidperoxide patterns in roots of Medicago truncatula during mycorrhizal . Levels of fungus-specific fatty acids as well as (16:0) and oleic (18:1 n - 9) were increased in mycorrhizal roots. Thus the degree of arbuscular mycorrhizal of roots can be estimated via analysis of fungal specific esterified fatty acids. Otherwise, no significant changes were found in the profiles of esterified and free fatty acids. The 9- and 13-LOX products of linoleic and alpha-linolenic were present in all root samples, but did not show significant differences between mycorrhizal and non-mycorrhizal roots, except JA which showed elevated levels in mycorrhizal roots. In both types of roots levels of 13-LOX products were higher than those of 9-LOX products. In addition, three cDNAs encoding CYP74 enzymes, two 9/13-hydroperoxide lyases and a 13-allene oxide synthase, were isolated and characterized. The transcript accumulation of these three genes, however, was not increased in mycorrhizal roots of M. truncatula.

Keyword: colonization

Phytoremediation assisted by mycorrhizal fungi of a Mexican defunct lead- battery recycling site.

A field experiment was conducted during 15\u202fmonths to study the effects of four arbuscular mycorrhizal fungi (AMF) on the growth of Ricinus communis accession SF7. Plants were established on amended soil (vermicompost:sawdust:soil 1:1:1) severely polluted by lead- batteries (LAB) located at Mexico State, Mexico. Plants inoculated with Acaulospora sp., Funneliformis mosseae and Gigaspora gigantea had 100% survival in comparison to non-inoculated plants (57%). These same AMF enhanced and linoleic acids content in seeds of R. communis. Acaulospora sp. modified rhizosphere soil pH and decreased 3.5 folds Pb foliar concentrations while F. mosseae BEG25 decreased three times Pb soil availability in comparison to non-inoculated plants. Spatial changes in Pb soil availability were observed at the end of this research. No fungal effect on P, Ca, Cu foliar concentrations, soluble sugars, proline, chlorophyll or on the activity of two oxidative stress enzymes was observed. Mycorrhizal from the inoculated fungi was between 40% and 60%, while by native fungi was between 16% and 22%. A similar percentage of foliar total phenolic compounds was observed in non-mycorrhizal plants and those inoculated with G. gigantea and Acaulospora sp. This is the first research reporting effects of AMF on R. communis (castor bean) shrubs when grown on a LAB recycling site suggesting the use of Acaulospora sp. and F. mosseae BEG25 in phytostabilization to ameliorate Pb pollution and decreasing its ecological risk.Copyright © 2018. Published by Elsevier B.V.

Keyword: colonization

New anti-infective coatings of surgical sutures based on a combination of antiseptics and fatty acids.

Wound infection is a complication feared in surgery. The aim of this study was to develop new anti-infective coatings of surgical sutures and to compare the anti-microbial effectiveness and biocompatibility to the well-established Vicryl Plus. Synthetic absorbable PGA surgical sutures were coated with three different chlorhexidine concentrations and two different octenidine concentrations in combination with and lauric . Drug-release kinetics lasting 96 h were studied in phosphate-buffered saline at 37 degrees C. Anti-infective characteristics were determined by measuring the change in optical density of Staphylococcus aureus suspensions charged with coated sutures over time. Microorganisms adsorbed at the surface of coated sutures were assessed on blood agar plates and coated sutures eluted for 24 h were placed on bacterial lawns cultured on Mueller-Hinton plates to prove retained anti-microbial potency. A cell proliferation assay was performed to assess the degree of cytotoxicity. Anti-infective characteristics and biocompatibility were compared to Vicryl Plus. A coating technology for slow-release drug-delivery systems on surgical sutures could be developed. All coatings showed a continuous drug release within 96 h. Individual chlorhexidine and octenidine coated sutures showed superior anti-infective characteristics but inferior biocompatibility in comparison to Vicryl Plus. We conclude that the developed anti-infective suture coatings consisting of lipid-based drug-delivery systems in combination with antiseptics are highly effective against bacterial in vitro; however, drug doses have to be adjusted to improve biocompatibility.

Keyword: colonization

ToxR regulates the production of lipoproteins and the expression of serum resistance in Vibrio cholerae.

The genes encoding three lipoproteins of Vibrio cholerae were identified by a combination of DNA sequence analysis and [3H]palmitate labeling of hybrid proteins encoded by TnphoA gene fusions. The expression of these three lipoproteins, TagA, AcfD, and TcpC, was controlled by ToxR, the cholera toxin transcriptional activator. The involvement of other bacterial lipoproteins in conferring resistance to the bactericidal effects of complement prompted us to examine this possibility in V. cholerae. Remarkably, mutations in toxR and tcp genes (including tcpC), involved in the biogenesis of the toxin coregulated pili, rendered V. cholerae about 10(4)-10(6) times more sensitive to the vibriocidal activity of antibody and complement. Since V. cholerae is a noninvasive organism and toxR and tcp mutants are highly defective in intestinal in animals and humans, these results raise the possibility that resistance to a gut-associated, "complement-like" bactericidal activity may be a major virulence determinant of V. cholerae and other enterobacterial species.

Keyword: colonization

Cell membrane-binding properties of group A streptococcal lipoteichoic .

Lipoteichoic (LTA) was extracted from group A streptococci, previously treated with hot HCl, by the phenol method. The extracted LTA was loaded on an isoelectric (IE) focusing column and two fractions were collected; one at pH 4.65 and the other at pH 2.95. Chemical analysis demonstrated that the unfractionated LTA contained alanine and glycerolphosphate at molar ratio of 1:10, and ester-linked lipids, but no detectable sugars or amino-sugars. The two IE fractions contained lipids but lacked alanine. The LTA and its IE fractions spontaneously adsorbed to human erythrocytes (sensitization) causing them to agglutinate in the presence of rabbit anti-LTA. The RBC-sensitizing and antigenic activities of IE fractions were equal to, or greater (for IE fraction at pH 4.65) than the unfractionated LTA, indicating that alanine is not involved in the sensitizing activity of LTA. Mild ammonia-hydrolysis abolished the RBC-sensitizing activity of LTA and its IE fractions. Chloroform-methanol-soluble material of the ammonia-hydrolysate lacked antigenic activity but blocked sensitization of erythrocytes by LTA. The water-soluble material of the hydrolyzed LTA retained antigenic activity, was not able to block sensitization by LTA, and its sensitizing activity was restored after esterification with fatty acids. These experiments indicate that ester-linked fatty acids ( being the major one) are involved in the spontaneous adsorption of LTA to erythrocytes. The LTA, its lipid moiety, and anti-LTA blocked adherence of group A streptococci to human epithelial cells, suggesting that small amounts of LTA may reside on the streptococcal surface to mediate attachment and of these organisms on mucosal surfaces in vivo.

Keyword: colonization

Adherence, coaggregation, and hydrophobicity of Streptococcus gordonii associated with expression of cell surface lipoproteins.

Streptococcus gordonii Challis incorporated exogenous [3H]palmitate into 13 polypeptides extractable from intact cells with sodium dodecyl sulfate. A 76-kDa surface-exposed polypeptide, implicated previously as a cell aggregation determinant, was shown to be one of these lipid-modified polypeptides. Differences in sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of lipopolypeptides were detected with mutants of S. gordonii that were altered in adherence, aggregation, coaggregation, or hydrophobicity. Lipid-modified polypeptides, tightly associated with the cell membrane, may be involved in the expression of cell surface properties of S. gordonii important for of the human oral cavity.

Keyword: colonization

Antiadhesive activity of the biosurfactant pseudofactin II secreted by the Arctic bacterium Pseudomonas fluorescens BD5.

Pseudofactin II is a recently identified biosurfactant secreted by Pseudomonas fluorescens BD5, the strain obtained from freshwater from the Arctic Archipelago of Svalbard. Pseudofactin II is a novel compound identified as cyclic lipopeptide with a connected to the terminal amino group of eighth amino in peptide moiety. The C-terminal carboxylic group of the last amino forms a lactone with the hydroxyl of Thr3. Adhesion is the first stage of biofilm formation and the best moment for the action of antiadhesive and anti-biofilm compounds. Adsorption of biosurfactants to a surface e.g. glass, polystyrene, silicone modifies its hydrophobicity, interfering with the microbial adhesion and desorption processes. In this study the role and applications of pseudofactin II as a antiadhesive compound has been investigated from medicinal and therapeutic perspectives.Pseudofactin II lowered the adhesion to three types of surfaces (glass, polystyrene and silicone) of bacterial strains of five species: Escherichia coli, Enterococcus faecalis, Enterococcus hirae, Staphylococcus epidermidis, Proteus mirabilis and two Candida albicans strains. Pretreatment of a polystyrene surface with 0.5 mg/ml pseudofactin II inhibited bacterial adhesion by 36-90% and that of C. albicans by 92-99%. The same concentration of pseudofactin II dislodged 26-70% of preexisting biofilms grown on previously untreated surfaces. Pseudofactin II also caused a marked inhibition of the initial adhesion of E. faecalis, E. coli, E. hirae and C. albicans strains to silicone urethral catheters. The highest concentration tested (0.5 mg/ml) caused a total growth inhibition of S. epidermidis, partial (18-37%) inhibition of other bacteria and 8-9% inhibition of C. albicans growth.Pseudofactin II showed antiadhesive activity against several pathogenic microorganisms which are potential biofilm formers on catheters, implants and internal prostheses. Up to 99% prevention could be achieved by 0.5 mg/ml pseudofactin II. In addition, pseudofactin II dispersed preformed biofilms. Pseudofactin II can be used as a disinfectant or surface coating agent against microbial of different surfaces, e.g. implants or urethral catheters.

Keyword: colonization

and mineralization of byPseudomonas pseudoflava.

Pseudomonas pseudoflava and were used to investigate the role of bacterial in the degradation of waterinsoluble organic compounds. Mineralization was measured by trapping the(14)CO2 produced from the labeled substrate, and of the surface of the solid organic chemical was determined by epifluorescence microscopy. In a medium containing solid ,P. pseudoflava mineralized the organic substrate at a logarithmic rate. Mineralization was evident before of the surface of the chemical was detected. The rate of appearance of single cells and/or aggregations of cells on the surface of the was essentially the same as the doubling time of free cells in solution. At about 50 hours, mineralization and of the surface stopped. In a salts solution containing solid ,P. pseudoflava grew logarithmically in the solution and biphasically on the surface of the . We suggest that the bacterium first metabolizes soluble and later colonizes the solid when the substrate in solution has been depleted.

Keyword: colonization

Docosahexaenoic and Eicosapentaenoic Acids Prevent Altered-Muc2 Secretion Induced by by Alleviating Endoplasmic Reticulum Stress in LS174T Goblet Cells.

Diets high in saturated fatty acids (FA) represent a risk factor for the development of obesity and associated metabolic disorders, partly through their impact on the epithelial cell barrier integrity. We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus barrier. To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or -9, -6, or -3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic nor -9 and -6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 and obesity or inflammatory bowel disease. These in vitro data remain to be explored in vivo in a context of dietary obesity.

Keyword: diabetes

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on lipid metabolism and inflammation in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular lipid contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet lipid metabolic disorder and islet inflammation in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

BH3 mimetics derived from Bim-BH3 domain core region show PTP1B inhibitory activity.

A series of our previously described BH3 peptide mimetics derived from Bim-BH3 domain core region were found to exhibit weak to potent PTP1B binding affinity and inhibitory activities via target-based drug screening. Among these compounds, a 12-aa Bim-BH3 core sequence peptide conjugated to (SM-6) displayed good PTP1B binding affinity (K\u202f=\u202f8.38\u202fnmol/L), inhibitory activity (IC\u202f=\u202f1.20\u202fμmol/L) and selectivity against other PTPs (TCPTP, LAR, SHP-1 and SHP-2). Furthermore, SM-6 promoted HepG2 cell glucose uptake and inhibited the expression of PTP1B, indicating that SM-6 could improve the insulin resistance effect in the insulin-resistant HepG2 cell model. These results may indicate a new direction for the application of BH3 peptide mimetics and promising PTP1B peptide inhibitors could be designed and developed based on SM-6.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Characterization of α-Glucosidase Inhibitors from Lindau Leaves by Gas Chromatography-Mass Spectrometry-Based Metabolomics and Molecular Docking Simulation.

() is an herbal shrub traditionally consumed to treat various diseases including in Malaysia. This study was designed to evaluate the α-glucosidase inhibitory activity of leaves extracts, and to identify the metabolites responsible for the bioactivity.Crude extract obtained from the dried leaves using 80% methanolic solution was further partitioned using different polarity solvents. The resultant extracts were investigated for their α-glucosidase inhibitory potential followed by metabolites profiling using the gas chromatography tandem with mass spectrometry (GC-MS).Multivariate data analysis was developed by correlating the bioactivity, and GC-MS data generated a suitable partial least square (PLS) model resulting in 11 bioactive compounds, namely, , phytol, hexadecanoic (methyl ester), 1-monopalmitin, stigmast-5-ene, pentadecanoic , heptadecanoic , 1-linolenoylglycerol, glycerol monostearate, alpha-tocospiro B, and stigmasterol. In-silico study via molecular docking was carried out using the crystal structure isomaltase (PDB code: 3A4A). Interactions between the inhibitors and the protein were predicted involving residues, namely LYS156, THR310, PRO312, LEU313, GLU411, and ASN415 with hydrogen bond, while PHE314 and ARG315 with hydrophobic bonding.The study provides informative data on the potential α-glucosidase inhibitors identified in leaves, indicating the plant\'s therapeutic effect to manage hyperglycemia.

Keyword: diabetes

Scopoletin intervention in pancreatic endoplasmic reticulum stress induced by lipotoxicity.

Endoplasmic reticulum (ER), a dynamic organelle, plays an essential role in organizing the signaling pathways involved in cellular adaptation, resilience, and survival. Impairment in the functions of ER occurs in a variety of nutritive disorders including obesity and type 2 . Here, we hypothesize that (scopoletin) SPL, a coumarin, has the potential to alleviate ER stress induced in vitro and in vivo models by lipotoxicity. To test this hypothesis, the ability of SPL to restore the levels of proteins of ER stress was analyzed. Rat insulinoma 5f (RIN5f) cells and Sprague Dawley rats were the models used for this study. Groups of control and high-fat, high-fructose diet (HFFD)-fed rats were treated with either SPL or 4-phenylbutyric . Status of ER stress was enumerated by quantitative RT-PCR, Western blot, electron microscopic, and immunohistochemical studies. Proximal proteins of ER stress inositol requiring enzyme 1 (IRE1), protein kinase like endoplasmic reticulum kinase (PERK), and activating transcription factor 6 (ATF6) were reduced in the β-cells by SPL. The subsequent signaling proteins X-box binding protein 1, eukaryotic initiation factor2α, activating transcription factor 4, and C/EBP homologous protein were also suppressed in their expression levels when treated with SPL. IRE1, PERK signaling leads to c-Jun-N-terminal kinases phosphorylation, a kinase that interrupts insulin signaling, which was also reverted upon scopoletin treatment. Finally, we confirm that SPL has the ability to suppress the stress proteins and limit pancreatic ER stress which might help in delaying the progression of insulin resistance.

Keyword: diabetes

The Mitochondria-Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

Mitochondria-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive oxygen species (ROS) production during glucose metabolism in β cells can be exacerbated under hyperglycaemic conditions such as type 2 (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

Palmitate is not an effective fuel for pancreatic islets and amplifies insulin secretion independent of calcium release from endoplasmic reticulum.

The aim of the study was to determine the acute contribution of fuel oxidation in mediating the increase in insulin secretion rate (ISR) in response to fatty acids. Measures of mitochondrial metabolism, as reflected by oxygen consumption rate (OCR) and cytochrome c reduction, calcium signaling, and ISR by rat islets were used to evaluate processes stimulated by acute exposure to (PA). The contribution of mitochondrial oxidation of PA was determined in the presence and absence of a blocker of mitochondrial transport of fatty acids (etomoxir) at different glucose concentrations. Subsequent to increasing glucose from 3 to 20 mM, PA caused small increases in OCR and cytosolic calcium (about 20% of the effect of glucose). In contrast, the effect of PA on ISR was almost 3 times that by glucose, suggesting that the metabolism of PA is not the dominant mechanism mediating PA\'s effect on ISR. This was further supported by lack of inhibition of PA-stimulated OCR and ISR when blocking entry of PA into mitochondria (with etomoxir), and PA\'s lack of stimulation of reduced cytochrome c in the presence of high glucose. Consistent with the lack of metabolic stimulation by PA, an inhibitor of calcium release from the endoplasmic reticulum, but not a blocker of L-type calcium channels, abolished the PA-induced elevation of cytosolic calcium. Notably, ISR was unaffected by thapsigargin showing the dissociation of endoplasmic reticulum calcium release and second phase insulin secretion. In conclusion, stimulation of ISR by PA was mediated by mechanisms largely independent of the oxidation of the fuel.

Keyword: diabetes

Docosahexaenoic Reduces -Induced Endoplasmic Reticulum Stress in Pancreatic Β Cells.

Endoplasmic reticulum (ER) stress leads to peripheral insulin resistance and the progression of pancreatic beta cell failure in type 2 . Although ER stress plays an important role in the pathogenesis of , it is indispensable for cellular activity. Therefore, when assessing the pathological significance of ER stress, it is important to monitor and quantify ER stress levels. Here, we have established a novel system to monitor ER stress levels quickly and sensitively, and using this method, we have clarified the effect of differences in glucose concentration and various fatty acids on the ER of pancreatic β cells. First, we developed a cell system that secretes Gaussia luciferase in culture medium depending on the activation of the GRP78 promoter. This system could sensitively monitor ER stress levels that could not be detected with real-time RT-PCR and immunoblotting. This system revealed that hyperglycemia does not induce unfolded protein response (UPR) in a short period of time in MIN6 cells, a mouse pancreatic β cell line. Physiological concentrations of , a saturated fatty , induced ER stress quickly, while physiological concentrations of oleic , an unsaturated fatty , did not. Docosahexaenoic , an n-3 unsaturated fatty , inhibited -induced ER stress. In this study, we have established a system that can sensitively detect ER stress levels of living cells in a short period of time. This system can be used to monitor the state of the ER in living cells and lead to the investigation of the significance of physiological or pathological ER stress levels.

Keyword: diabetes

The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis.

Hyperglycemia and hyperlipidemia (glycolipotoxicity)-triggered islet β-cell dysfunction is known to drive the progression of obesity-related type 2 , however the underlying mechanisms have not been clearly elucidated. The current study aimed to investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP-5) in islet cells under glucolipotoxic conditions. Using gene overexpression and knockdown approaches, we demonstrated that MKP-5 could alleviate glucolipotoxicity-induced apoptosis via the endoplasmic reticulum (ER) stress and mitochondrial apoptosis pathways owing to the altered regulation of caspase family members and ER stress-related molecules in MIN6 and primary islet cells. Overexpression of MKP-5 reversed the glucose and (GP)-induced impairment of insulin secretion as well as the abnormal decreases in the expression of islet functional genes, thereby maintaining the normal insulin secretory functionality, whereas the absence of MKP-5 aggravated islet cell dysfunction. In parallel, the production of ROS and increased inflammation-associated genes in response to GP were also reduced upon MKP-5 overexpression. Further, inhibition of JNK or P38 MAPK pathways resisted to glucolipotoxicity observed in MKP-5 knockdown MIN6 cells. These findings indicate that MKP-5 is an important mediator for glucolipotoxicity-induced islet cell dysfunction and apoptosis, with JNK and P38 as the critical downstream pathways.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Probing the Global Cellular Responses to Lipotoxicity Caused by Saturated Fatty Acids.

Excessive levels of saturated fatty acids are toxic to cells, although the basis for this lipotoxicity remains incompletely understood. Here, we analyzed the transcriptome, lipidome, and genetic interactions of human leukemia cells exposed to palmitate. Palmitate treatment increased saturated glycerolipids, accompanied by a transcriptional stress response, including upregulation of the endoplasmic reticulum (ER) stress response. A comprehensive genome-wide short hairpin RNA (shRNA) screen identified >350 genes modulating lipotoxicity. Among previously unknown genetic modifiers of lipotoxicity, depletion of RNF213, a putative ubiquitin ligase mutated in Moyamoya vascular disease, protected cells from lipotoxicity. On a broader level, integration of our comprehensive datasets revealed that changes in di-saturated glycerolipids, but not other lipid classes, are central to lipotoxicity in this model. Consistent with this, inhibition of ER-localized glycerol-3-phosphate acyltransferase activity protected from all aspects of lipotoxicity. Identification of genes modulating the response to saturated fatty acids may reveal novel therapeutic strategies for treating metabolic diseases linked to lipotoxicity.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 .

Metabolic stress-induced low-grade chronic inflammation plays an important role in the development of insulin-resistance and type 2 (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the obesity-induced metabolic stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: diabetes

A Novel Role for Somatostatin in the Survival of Mouse Pancreatic Beta Cells.

Cross-talk between different pancreatic islet cell types regulates islet function and somatostatin (SST) released from pancreatic delta cells inhibits insulin secretion from pancreatic beta cells. In other tissues SST exhibits both protective and pro-apoptotic properties in a tissue-specific manner, but little is known about the impact of the peptide on beta cell survival. Here we investigate the specific role of SST in the regulation of beta cell survival in response to physiologically relevant inducers of cellular stress including palmitate, cytokines and glucose.Pancreatic MIN6 beta cells and primary mouse islet cells were pre-treated with SST with or without the G signalling inhibitor, pertussis toxin, and exposed to different cellular stress factors. Apoptosis and proliferation were assessed by measurement of caspase 3/7 activity, TUNEL and BrdU incorporation, respectively, and expression of target genes was measured by qPCR.SST partly alleviated upregulation of cellular stress markers (Hspa1a and Ddit3) and beta cell apoptosis in response to factors such as lipotoxicity (palmitate), pro-inflammatory cytokines (IL1β and TNFα) and low glucose levels. This effect was mediated via a G protein-dependent pathway, but did not modify transcriptional upregulation of the specific NFκB-dependent genes, Nos2 and Ccl2, nor was it associated with transcriptional changes in SST receptor expression.Our results suggest an underlying protective effect of SST which modulates the beta cell response to ER stress and apoptosis induced by a range of cellular stressors associated with type 2 .© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

Mitochondria-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.

Can we use a mitochondrial-targeted antioxidant (Mitoquinone) during in vitro embryo culture to rescue developmental competence of oocytes matured under lipotoxic conditions, exhibiting mitochondrial dysfunction and oxidative stress?Supplementation of embryo culture media with Mitoquinone reduced oxidative stress and prevented mitochondrial uncoupling in embryos derived from metabolically compromised oocytes in vitro, leading to higher blastocyst rates and lower blastomeric apoptosis.Maternal metabolic disorders, such as obesity and type-II are associated with hyperlipidemia and elevated free fatty (FFA) concentrations in the ovarian follicular fluid (FF). Oocyte maturation under these lipotoxic conditions results in increased oxidative stress levels, mitochondrial dysfunction, reduced developmental competence and disappointing IVF results.A well-described bovine oocyte IVM model was used, where a pathophysiologically relevant elevated FF concentrations of (PA; 150\xa0μM or 300\xa0μM) were added to induce oxidative stress. After fertilization (Day 0, D0), zygotes were in vitro cultured (IVC, from D1 to D8) in standard fatty -free media in the presence or absence of Mitoquinone or its carrier triphenyl-phosphonium.Embryo cleavage and fragmentation (D2) and blastocyst rates (D8) were recorded. Mitochondrial activity and oxidative stress in cleaved embryos at D2 were determined using specific fluorogenic probes and confocal microscopy. D8 blastocysts were used to (i) examine the expression of marker genes related to mitochondrial unfolded protein responses (UPRmt; HSPD1 and HSPE1), mitochondrial biogenesis (TFAM), endoplasmic reticulum (ER) UPR (ATF4, ATF6 and BiP) and oxidative stress (CAT, GPX1 and SOD2) using real time RT-PCR; (ii) determine cell differentiation and apoptosis using CDX-2 and cleaved caspase-3 immunostaining; and (iii) measure mtDNA copy numbers. This was tested in a series of experiments with at least three independent replicates for each, using a total of 2525 oocytes. Differences were considered significant if a P value was <0.05 after Bonferroni correction.Exposure to PA during IVM followed by culture under control conditions resulted in a significant increase in oxidative stress in embryos at D2. This was associated with a significant reduction in mitochondrial inner membrane potential (uncoupling) compared with solvent control (P\xa0<\u20090.05). The magnitude of these effects was PA-concentration dependent. Consequently, development to the blastocysts stage was significantly hampered. Surviving blastocysts exhibited high apoptotic cell indices and upregulated mRNA expression indicating persistent oxidative stress, mitochondrial and ER UPRs. In contrast, supplementation of PA-derived zygotes with Mitoquinone during IVC (i) prevented mitochondrial uncoupling and alleviated oxidative stress at D2; and (ii) rescued blastocyst quality; normalized oxidative stress and UPR related genes and apoptotic cell indices (P\xa0>\u20090.01 compared with solvent control). Mitoquinone also improved blastocyst rate in PA-exposed groups, an effect that was dependent on PA concentration.N/A.This is a fundamental study performed using a bovine in vitro model using PA-induced lipotoxicity during oocyte maturation. PA is the most predominant FFA in the FF that is known to induce lipotoxicity; however, in vivo maturation in patients suffering from maternal metabolic disorders involve more factors that cannot be represented in one model. Nevertheless, focusing on the carryover oxidative stress as a known key factor affecting developmental competence, and considering the novel beneficial rescuing effects of Mitoquinone shown here, we believe this model is of high biological relevance.Human oocytes collected for IVF treatments from patients with maternal metabolic disorders are vulnerable to lipotoxicity and oxidative stress during in vivo maturation. The results shown here suggest that mitochondrial targeted therapy, such as using Mitoquinone, during IVC may rescue the developmental competence and quality of these compromised oocytes. After further clinical trials, this may be a valuable approach to increase IVF success rates for infertile patients experiencing metabolic disorders.This study was financially supported by a BOF/KP grant number 34399, from the University of Antwerp, Belgium. W.F.A.M. was supported by a postdoctoral fellowship from the Research Foundation-Flanders (FWO), grant number 12I1417N, Antwerp, Belgium. The Leica SP 8 confocal microscope used in this study was funded by the Hercules Foundation of the Flemish Government (Hercules grant AUHA.15.12). All authors have no financial or non-financial competing interests to declare.© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: diabetes

Glycation of human serum albumin impairs binding to the glucagon-like peptide-1 analogue liraglutide.

The long-acting glucagon-like peptide-1 analogue liraglutide has proven efficiency in the management of type 2 and also has beneficial effects on cardiovascular diseases. Liraglutide\'s protracted action highly depends on its capacity to bind to albumin via its part. However, in , albumin can undergo glycation, resulting in impaired drug binding. Our objective in this study was to assess the impact of human serum albumin (HSA) glycation on liraglutide affinity. Using fluorine labeling of the drug and F NMR, we determined HSA affinity for liraglutide in two glycated albumin models. We either glycated HSA by incubation with glucose (G25- or G100-HSA) or methylglyoxal (MGO-HSA) or purified glycated HSA from the plasma of diabetic patients with poor glycemic control. Nonglycated commercial HSA (G0-HSA) and HSA purified from plasma of healthy individuals served as controls. We found that glycation decreases affinity for liraglutide by 7-fold for G100-HSA and by 5-fold for MGO-HSA compared with G0-HSA. A similarly reduced affinity was observed for HSA purified from diabetic individuals compared with HSA from healthy individuals. Our results reveal that glycation significantly impairs HSA affinity to liraglutide and confirm that glycation contributes to liraglutide\'s variable therapeutic efficiency, depending on stage. Because is a progressive disease, the effect of glycated albumin on liraglutide affinity found here is important to consider when is managed with this drug.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: diabetes

Mangiferin Improved Palmitate-Induced-Insulin Resistance by Promoting Free Fatty Metabolism in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved Insulin Resistance.

Elevated free fatty (FFA) is a key risk factor for insulin resistance (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high-fat diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA metabolism in HepG2 and C2C12 cells. The model was used to quantify PA-induced lipid accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased insulin-stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA metabolism were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: diabetes

Transport of Ca and Ca-Dependent Permeability Transition in Rat Liver Mitochondria under the Streptozotocin-Induced Type I .

Although is known to be a disease associated with mitochondrial dysfunction, not everything is clear about mitochondrial Ca transport and Ca-induced permeability transition in diabetic cells. The objective of this work was to study the operation of MCU and Ca-dependent mitochondrial permeabilization in the liver cells of Sprague-Dawley rats under the streptozotocin-induced type I . It was shown that two weeks after the induction of , the rate of Ca uptake by the mitochondria of diabetic animals increased ~1.4-fold. The expression of MCU and MICU1 subunits did not change, yet the quantity of dominant-negative MCUb channel subunits was almost twice as lower. The organelles also became more resistant to the induction of CsA-sensitive MPT pore and less resistant to the induction of CsA-insensitive palmitate/Ca-induced pore. The mitochondria of diabetic liver cells also showed changes in the lipid matrix of their membranes. The content of fatty acids in the membranes grew, and microviscosity of the lipid bilayer (assessed with laurdan) increased. At the same time, lipid peroxidation (assessed by the production of malonic dialdehyde) was stimulated. The paper discusses the consequences of the -related changes in mitochondria in the context of cell physiology.

Keyword: diabetes

Taurine improves low-level inorganic arsenic-induced insulin resistance by activating PPARγ-mTORC2 signalling and inhibiting hepatic autophagy.

Inorganic arsenic (iAs) is reportedly associated with the increased incidence of type 2 in the population. Here, we found that iAs exposure significantly decreased the expression of glycolytic genes and glycogen content and increased gluconeogenesis gene levels in C57/BL6J mice. The expression of peroxisome proliferator-activated receptor γ (PPARγ), and mechanistic target of rapamycin complex 2 (mTORC2) were decreased in the livers of iAs-treated mice. Furthermore, in iAs-treated HepG2 cells, we found that PPARγ agonist rosiglitazone (RGS) increased the expression of mTORC2, inhibited autophagy, and improved glucose metabolism. mTORC2 agonist inhibited autophagy and improved glucose metabolism as well as the autophagosome formation inhibitor 3-methyladenine. Taurine, a natural compound, reversed impaired glucose metabolism and decreased expression of PPARγ and mTORC2 induced by iAs in mice liver and HepG2 cells. These data indicated that taurine administration could ameliorate iAs-induced insulin resistance through activating PPARγ-mTORC2 signalling and subsequently inhibiting hepatic autophagy.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Triterpenoids Improve -Induced Hepatic Inflammation the Rho-Kinase-Dependent Pathway.

This study aimed to assess the effects of triterpene extract of (Batal.) Iljinskaja (CPT) on -induced hepatic inflammation and to unveil the underlying mechanisms. in db/db mice was alleviated after CPT administration, as assessed by the oral glucose tolerance test. In addition, treatment with CPT dramatically reduced serum insulin, aspartate amino-transaminase, alanine aminotransferase, triglyceride, and total cholesterol amounts. Besides, serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were also reduced after CPT administration. Western blot analysis revealed that CPT treatment significantly reversed the protein expression levels of Rho, ROCK1, ROCK2, p-P65, p-IκBα, p-IKKα, and p-IKKβ in liver samples obtained from db/db mice. Upon stimulation, the protective effects of CPT on the liver were further assessed in HepG2 and LO2 cells, and no appreciable cytotoxic effects were found. Therefore, these findings indicate that CPT alleviates liver inflammation Rho-kinase signaling. Metformin (PubChem CID: 4091); Fasudil (PubChem CID: 3547); (PubChem CID: 985).

Keyword: diabetes

Histone demethylase UTX is a therapeutic target for diabetic kidney disease.

Diabetic kidney disease (DKD) is a major complication of . We found that UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase, was upregulated in the renal mesangial and tubular cells of diabetic mice and DKD patients. In cultured renal mesangial and tubular cells, UTX overexpression promoted -induced elevation of inflammation and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. We found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes and apoptosis; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the -induced renal abnormalities in db/db mice, an animal model of type 2 . These results revealed the possible mechanisms underlying the regulation of histone methylation in DKD and suggest UTX as a potential therapeutic target for DKD.Diabetic kidney disease (DKD) is a microvascular complication of and the leading cause of end-stage kidney disease worldwide without effective therapy available. UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase that removes the di- and tri-methyl groups from histone H3K27, plays important biological roles in gene activation, cell fate control and life span regulation in Caenorhabditis elegans. In the present study, we report upregulated UTX in the kidneys of diabetic mice and DKD patients. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the -induced renal dysfunction, abnormal morphology, inflammation, apoptosis and DNA damage in db/db mice, comprising an animal model of type 2 . In cultured renal mesanglial and tubular cells, UTX overexpression promoted induced elevation of inflammation and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. Mechanistically, we found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Collectively, our results suggest UTX as a potential therapeutic target for DKD.© 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

Keyword: diabetes

Phytochemical Profiling, GC-MS Analysis and α-Amylase Inhibitory Potential of Ethanolic Extract of Linn. Endocarp.

Drugs with post-prandial action constitute one of the main courses of treatments for .In the present investigation, we have explored the α-amylase inhibitory potential of ethanolic extract of Cocos nucifera endocarp.DNS based assay was done to assess the α-amylase inhibition potential of ethanolic extract. Phytochemical screening and GC-MS analysis were done in order to assess the chemical profiling of extract. In silico docking studies were done using VLife MDS 4.6 software and the probable molecules, predicted after GC-MS analysis, were docked with the co-crystallized (acarbose) tracked active site and rest all cavities of porcine pancreatic α-amylase (1OSE). ADMET analysis was done using StarDrop 6.4, Derek Nexus and P450 Modules from Optibrium Ltd. and LHASA Ltd.DNS based α-amylase assay indicated that the IC50 value of extract lies in the range of 63- 126 µg/ml and at higher doses, i.e. above 250 µg/ml, it has better α-amylase inhibition than the standard drug, acarbose. Phytochemical screening indicated that ethanolic extract is rich in alkaloids, tannins, flavonoids, saponins, triterpenes, glycosides, carbohydrates, terpenoids, quinones and lactones. Further, GC-MS analysis (where Similarity Index was > 90) predicted that the probable phytoconstituents present in the ethanolic extract are myristic , syringaldehyde, eugenol, vanillin, 2,4-di-tert-butylphenol, lauric , methyl ester and γ-sitosterol. γ-Sitosterol showed the strong affinity towards the active site which was tracked by a co-crystallized ligand along with cavity 1 and 2 while significant interactions were observed in case of co-crystallized tracked active site as well as cavity 4 of 1OSE. Ethanolic extract of C. nucifera has no hemolytic effect.Its ability to effectively inhibit α-amylase may be attributed to the presence of the above probable molecules, which will be explored further.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: diabetes

Vascular endothelial growth factor over-expressed mesenchymal stem cells-conditioned media ameliorate palmitate-induced diabetic endothelial dysfunction through PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathway.

In the pathogenesis of (DM), islet microvasculares are severely damaged due to glucolipotoxicity and other reasons. Vascular endothelial growth factor (VEGF) is an indispensable and specific angiogenic factor in the pathogenesis and treatment of diabetic islet microvascular disease. Mesenchymal stem cells (MSCs) are regarded as a promising treatment of because of their immunosuppressive effect and multipotential differentiation potency. In this study, we tested whether MSCs over-expressing VEGF conditioned medium (MSC-VEGF-CM) could ameliorate pancreatic islet endothelial cells (MS-1) dysfunction induced by a common diabetic inducer palmitate (PA). We found that cell survival and migration were restrained by PA and partly repaired by the pro-protected of MSC-VEGF-CM. Meanwhile, PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathways were also up-regulated. Though apoptosis-related proteins, caspase-3 and caspase-9, had no significantly suppressed between MSC-VEGF-CM and MSC-CM alone, the expression levels of vascular surface factors such as CD31, VE-cadherin, occludin and ICAM-1, were remarkably up-regulated by the pro-protected of MSC-VEGF-CM. Our data suggested that MSC-VEGF-CM had therapeutic effect on the PA-induced dysfunction through the re-activation of PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathways.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Systematic review of palm oil consumption and the risk of cardiovascular disease.

The high amount of saturated fatty acids (SFA) coupled with the rising availability and consumption of palm oil have lead to the assumption that palm oil contributes to the increased prevalence of cardiovascular diseases worldwide. We aimed at systematically synthesising the association of palm oil consumption with cardiovascular disease risk and cardiovascular disease-specific mortality.We systematically searched Central, Medline and Embase databases up to June 2017 without restriction on setting or language. We performed separate searches based on the outcomes: coronary heart disease and stroke, using keywords related to these outcomes and palm oil. We searched for published interventional and observational studies in adults (Age: >18 years old). Two investigators extracted data and a consensus was reached with involvement of a third. Only narrative synthesis was performed for all of the studies, as the data could not be pooled.Our search retrieved 2,738 citations for stroke with one included study and 1,777 citations for coronary heart disease (CHD) with four included studies. was reported to be associated with risk of myocardial infarction (MI) (OR 2.76; 95%CI = 1.39-5.47). Total SFA intake was reported to be not significant for risk of MI. Varying intake of fried foods, highest contributor to total SFA with 36% of households using palm oil for frying, showed no significant associations to risk of MI. Odds of developing first non-fatal acute MI was higher in palm oil compared to soybean oil with 5% trans-fat (OR = 1.33; 95%CI = 1.09-1.62) than palm oil compared to soybean oil with 22% trans-fat (OR = 1.16; 95%CI = 0.86-1.56). Nevertheless, these risk estimates were non-significant and imprecise. The trend amongst those taking staple pattern diet (characterised by higher palm oil, red meat and added sugar consumption) was inconsistent across the factor score quintiles. During the years of 1980 and 1997, for every additional kilogram of palm oil consumed per-capita annually, CHD mortality risk was 68 deaths per 100,000 (95% CI = 21-115) in developing countries and 17 deaths per 100,000 (95%CI = 5.3-29) in high-income countries, whereas stroke was associated with 19 deaths per 100,000 (95%CI = -12-49) and 5.1 deaths per 100,000 (95% CI: -1.2-11) respectively. The evidence for the outcomes of this review were all graded as very low. The findings of this review should be interpreted with some caution, owing to the lack of a pooled effect estimate of the association, significant bias in selection criteria and confounding factors, inclusion of other food items together with palm oil, and the possible out-dated trend in the ecological study.In view of the abundance of palm oil in the market, quantifying its true association with CVD outcomes is challenging. The present review could not establish strong evidence for or against palm oil consumption relating to cardiovascular disease risk and cardiovascular disease-specific mortality. Further studies are needed to establish the association of palm oil with CVD. A healthy overall diet should still be prioritised for good cardiometabolic health.

Keyword: diabetes

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced . However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: diabetes

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance.

Patients with type 2 (T2D) and/or insulin resistance (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the insulin resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased insulin-stimulated glucose and FA uptake, as well as loss of endosomal acidification and insulin signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to obesity, and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: diabetes

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 . However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut microbiota and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: diabetes

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: diabetes

Identifying Signalling Pathways Regulated by GPRC5B in β-Cells by CRISPR-Cas9-Mediated Genome Editing.

CRISPR-Cas9, a RNA-guided targeted genome editing tool, has revolutionized genetic engineering by offering the ability to precisely modify DNA. GPRC5B is an orphan receptor belonging to the group C family of G protein-coupled receptors (GPCRs). In this study, we analysed the functional roles of the Gprc5b receptor in MIN6 β-cells using CRISPR-Cas9 and transient over-expression of Gprc5b.The optimal transfection reagent for use in MIN6 β-cells was determined by analysing efficiency of GFP plasmid delivery by cell sorting. A MIN6 β-cell line in which Gprc5b expression was knocked down (Gprc5b KD) was generated using CRISPR-Cas9 technology. Gprc5b receptor mRNA expression, proliferation, apoptosis, Cignal 45-Pathway Reporter Array signalling and western blot assays were carried out using Gpcr5b KD MIN6 β-cells that had been transiently transfected with different concentrations of mouse Gprc5b plasmid to over-express Gprc5b.JetPRIME® was the best candidate for MIN6 β-cell transfection, providing approximately 30% transfection efficiency. CRISPR-Cas9 technology targeting Gprc5b led to stable knock-down of this receptor in MIN6 β-cells and its re-expression induced proliferation and potentiated cytokine- and palmitate-induced apoptosis. The Cignal 45 Reporter analysis indicated Gprc5b-dependent regulation of apoptotic and proliferative pathways, and western blotting confirmed activation of signalling via TGF-β and IFNγ.This study provides evidence of CRISPR-Cas9 technology being used to down-regulate Gprc5b expression in MIN6 β-cells. This strategy allowed us to identify signalling pathways linking GPRC5B receptor expression to β-cell proliferation and apoptosis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Telmisartan protects against high glucose/high lipid-induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of . In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of . In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: diabetes

CTRP3 attenuates high-fat diet-induced male reproductive dysfunction in mice.

Recent studies have suggested a role for abdominal obesity in male infertility. Previous studies have found that cell apoptosis exerts an important role in obesity-related male infertility. C1q/TNF-related protein 3 (CTRP3), a paralog of adiponectin, has been proposed to exert anti-apoptotic effects and to attenuate -related cardiac injuries. However, the role of CTRP3 in high-fat diet (HFD)-induced spermatogenic impairment remains unclear. In the present study, we fed male mice an HFD for 24 weeks to induce obesity. The expression of CTRP3 was decreased by HFD feeding. Supplementation with the recombinant human globular domain of CTRP3 (0.25 μg/g/day) for 4 weeks beginning at 20 weeks of the HFD improved spermatogenic function in the HFD-fed mice, which were characterized by improved testis morphology, increased testis weight/body weight ratio, and increased sperm count, sperm viability, and sperm motility. We also found that CTRP3 infusion resulted in the attenuation of endoplasmic reticulum (ER) stress and the activation of silence information regulator 1 (SIRT1) in the testes of obese mice. Our study also suggested that CTRP3 attenuated the (PA)-induced reductions in sperm viability and motility via the inhibition of ER stress. Moreover, germ cell-specific knockout abolished the protective effects of CTRP3 and studies of human sperm showed that the protective effects of CTRP3 on sperm viability and motility were abrogated by a specific inhibitor of SIRT1. Thus, our results demonstrated that CTRP3 expression protected against HFD-induced spermatogenic deficiency through the SIRT1/ER stress pathway.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I , 2. rare structurally-related (receptor pathology) hyperinsulinemic type II , and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: diabetes

Microfluidic-enabled quantitative measurements of insulin release dynamics from single islets of Langerhans in response to 5- hydroxy stearic .

Proper release of insulin from pancreatic islets of Langerhans is essential for maintaining glucose homeostasis. For full efficacy, both the pattern and the amount of hormone release are critical. It is therefore important to understand how insulin levels are secreted from single islets in both a quantitative fashion and in a manner that resolves temporal dynamics. In this study, we describe a microfluidic analytical system that can both quantitatively monitor insulin secretion from single islets while simultaneously maintaining high temporal sampling to resolve dynamics of release. We have applied this system to determine the acute and chronic effects of a recently-identified lipid, 5- hydroxy stearic (5-PAHSA), which is a member of the fatty hydroxy fatty class of lipids that are upregulated in healthy individuals. Chronic incubation (48 h) with 5-PAHSA significantly increased glucose-stimulated insulin secretion (GSIS) in murine islets compared to chronic incubation without the lipid or in the presence of (PA). The studies were continued in human islets from both healthy donors and donors diagnosed with type 2 (T2DM). Total amounts of GSIS were not only augmented in islets that were chronically incubated with 5-PAHSA, but the dynamic insulin release profiles also improved as noted by more pronounced insulin oscillations. With this quantitative microfluidic system, we have corroborated the anti-diabetic effects of 5-PAHSA by demonstrating improved islet function after chronic incubation with this lipid via improved oscillatory dynamics along with higher basal and peak release rates.

Keyword: diabetes

Antioxidant and antidiabetic activity of blackberry after gastrointestinal digestion and human gut microbiota fermentation.

Blackberry fruit contains high levels of polyphenols particularly anthocyanins which contribute to its biological activities. Bioavailability of polyphenols especially anthocyanins is generally low, it has been proposed that metabolites from polyphenol biotransformation under colonic fermentation are components that exert health benefits. In this study, blackberry was subjected to simulated gastrointestinal digestion and gut microbiota fermentation at different time intervals (0-48\u202fh) to study the changes in bioactive components, its antioxidant and antidiabetic activities. Phenolic compounds, during digestion and fermentation were also analysed. Gut metabolites of blackberry significantly increased the glucose consumption and glycogen content in HepG2 cells. Furthermore, gut metabolites ameliorated high glucose plus -induced ROS overproduction, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. The mechanism of antidiabetic activity of blackberry was via its potent antioxidant activity. Therefore, our results suggest that blackberry could be recommended as a functional food due to potential antioxidant and antidiabetic activity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Islet protection and amelioration of type 2 by treatment with quercetin from the flowers of .

The traditional Chinese medicine - the flower of - is reported as an effective therapeutic for type 2 (T2DM). Nevertheless, most constituents of the flowers of have not yet been studied. This study was conducted to investigate the effect of quercetin extracted from the flowers of on islet protection and amelioration in T2DM and explore its mechanism.Quercetin was extracted from the flowers of and verified by high-performance liquid chromatography. Quercetin or crude extract\'s effect on insulin secretion was investigated. ERK1/2 and phospho-ERK1/2 were detected by Western blot analysis, and fluo-3 AM was used to detect intracellular Ca. The anti-apoptosis effect of quercetin or crude extract on MIN-6 cells was investigated by thiazolyl blue tetrazolium bromide (MTT) assay and flow cytometry analysis. Activation of caspases and expression of Bcl-2 and BAX were tested by Western blot analysis. In addition, the mitochondrial membrane potential was determined by JC-1 probe. Moreover, in vivo activity was also tested in db/db mice.A quercetin level of >10 μmol/L could induce insulin secretion. Intracellular Ca and ERK1/2 were involved in the signaling pathway of quercetin-induced insulin secretion. We also observed that quercetin could inhibit -induced cell apoptosis via suppressing the activation of caspase-3, -9, -12; increasing the ratio of Bcl-2/BAX and reversing the impaired mitochondrial membrane potential. Crude extract\'s effect on insulin secretion was similar to that of pure extracted quercetin, while it possessed higher anti-apoptosis activity. Additionally, intraperitoneal glucose tolerance, plasma insulin level, hepatic triglyceride, hepatic glycogen and the pathological histology of both pancreatic islet and liver in db/db mice were significantly improved by the administration of the extracted quercetin.Our study indicated that quercetin extracted from the flowers of exerted excellent properties in islet protection and amelioration.

Keyword: diabetes

Downregulation of MALAT1 alleviates saturated fatty -induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis.

The increased level of saturated fatty acids (SFAs) is found in patients with , obesity, and other metabolic disorders. SFAs can induce lipotoxic damage to cardiomyocytes, but the mechanism is unclear. The long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acts as a key regulator in (PA)-induced hepatic steatosis, but its role in PA-induced myocardial lipotoxic injury is still unknown. The aim of this study was to explore the role and underlying mechanism of MALAT1 in PA-induced myocardial lipotoxic injury. MALAT1 expression in PA-treated human cardiomyocytes (AC16 cells) was detected by RT-qPCR. The effect of MALAT1 on PA-induced myocardial injury was measured by Cell Counting Kit-8, lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) assays. Apoptosis was detected by flow cytometry. The activities of cytokines and nuclear factor (NF)-κB were detected by enzyme-linked immunosorbent assay. The interaction between MALAT1 and miR-26a was evaluated by a luciferase reporter assay and RT-qPCR. The regulatory effects of MALAT1 on high mobility group box 1 (HMGB1) expression were evaluated by RT-qPCR and western blotting. MALAT1 was significantly upregulated in cardiomyocytes after PA treatment. Knockdown of MALAT1 increased the viability of PA-treated cardiomyocytes, decreased apoptosis, and reduced the levels of LDH, CK-MB, TNF-α, and IL-1β. Moreover, we found that MALAT1 specifically binds to miR-26a and observed a reciprocal negative regulatory relationship between these factors. We further found that the downregulation of MALAT1 represses HMGB1 expression, thereby inhibiting the activation of the Toll-like receptor 4 (TLR4)/NF-κB-mediated inflammatory response. These repressive effects were rescued by an miR-26a inhibitor. We demonstrate that MALAT1 is induced by SFAs and its downregulation alleviates SFA-induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis. Our findings provide new insight into the mechanism underlying myocardial lipotoxic injury.

Keyword: diabetes

High-density lipoprotein ameliorates -induced lipotoxicity and oxidative dysfunction in H9c2 cardiomyoblast cells via ROS suppression.

High levels circulating saturated fatty acids are associated with , obesity and hyperlipidemia. In heart, the accumulation of saturated fatty acids has been determined to play a role in the development of heart failure and diabetic cardiomyopathy. High-density lipoprotein (HDL) has been reported to possess key atheroprotective biological properties, including cellular cholesterol efflux capacity, anti-oxidative and anti-inflammatory activities. However, the underlying mechanisms are still largely unknown. Therefore, the aim of the present study is to test whether HDL could protect (PA)-induced cardiomyocyte injury and explore the possible mechanisms.H9c2 cells were pretreated with HDL (50-100\u2009μg/ml) for 2\u2009h followed by PA (0.5\u2009mM) for indicated time period. Our results showed that HDL inhibited PA-induced cell death in a dose-dependent manner. Moreover, HDL rescued PA-induced ROS generation and the phosphorylation of JNK which in turn activated NF-κB-mediated inflammatory proteins expressions. We also found that PA impaired the balance of BCL family proteins, destabilized mitochondrial membrane potential, and triggered subsequent cytochrome c release into the cytosol and activation of caspase 3. These detrimental effects were ameliorated by HDL treatment.PA-induced ROS accumulation and results in cardiomyocyte apoptosis and inflammation. However, HDL attenuated PA-induced lipotoxicity and oxidative dysfunction via ROS suppression. These results may provide insight into a possible molecular mechanism underlying HDL suppression of the free fatty -induced cardiomyocyte apoptosis.

Keyword: diabetes

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and lipid metabolism.

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet lipid metabolism in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high-fat diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced lipid accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced lipid deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell lipid accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

Role of the saturated fatty palmitate in the interconnected hypothalamic control of energy homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body energy and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, obesity, and type 2 . Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and energy homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: diabetes

Neuroprotective effects of vitamin D on high fat diet- and -induced enteric neuronal loss in mice.

The role of vitamin D in obesity and is debated. Obese and/or diabetic patients have elevated levels of free fatty acids, increased susceptibility to gastrointestinal symptoms and are suggested to have altered vitamin D balance. The enteric nervous system is pivotal in regulating gastrointestinal activity and high fat diet (HFD) has been shown to cause loss of enteric neurons in ileum and colon. This study investigates the effect of vitamin D on HFD- and -induced enteric neuronal loss in vivo and in vitro.Mice were fed either a normal diet (ND) or HFD supplemented with varying levels of vitamin D (from 0x to 20x normal vitamin D level) for 19 weeks. Ileum and colon were analyzed for neuronal numbers and remodeling. Primary cultures of myenteric neurons from mouse small intestine were treated with (4x10M) and/or 1α,25-hydroxy-vitamin D3 (VD, 10- 10M) with or without modulators of lipid metabolism and VD pathways. Cultures were analyzed by immunocyto- and histochemical methods.Vitamin D supplementation had no effect on enteric neuronal survival in the ND group. HFD caused substantial loss of myenteric neurons in ileum and colon. Vitamin D supplementation between 0-2x normal had no effect on HFD-induced neuronal loss. Supplementation with 20x normal, prevented the HFD-induced neuronal loss. In vitro supplementation of VD prevented the -induced neuronal loss. The VD receptor (VDR) was not identified in enteric neurons. Enteric glia expressed the alternative VD receptor, protein disulphide isomerase family A member 3 (PDIA3), but PDIA3 was not found to mediate the VD response in vitro. Inhibition of peroxisome proliferator-activated receptor gamma (PPARγ) and immune neutralization of isocitrate lyase prevented the VD mediated neuroprotection to exposure.Results show that VD protect enteric neurons against HFD and induced neuronal loss. The mechanism behind is suggested to be through activation of PPARγ leading to improved neuronal peroxisome function and metabolism of neuronal lipid intermediates.

Keyword: diabetes

Protective potential of klotho protein on diabetic retinopathy: Evidence from clinical and in\xa0vitro studies.

The purpose of the present study was to observe the relationship between serum α-klotho (KL) protein level and diabetic retinopathy (DR), and to further examine the effects of KL protein on apoptosis induced by (PA) in human retinal endothelial cells.A total of 17 healthy people and 60 type\xa02 patients were included. According to the results from fundus fluorescein angiography, the patients were divided into three subgroups: without DR, non-proliferative DR and proliferative DR. Serum KL level was measured by enzyme-linked immunosorbent assay. In\xa0vitro, human retinal endothelial cells were exposed to PA with or without KL protein. Apoptosis rates were analyzed by flow cytometry analysis. Apoptotic-related protein expressions were detected by western blotting analysis.Serum KL level was lower in patients than that in healthy participants (P\xa0=\xa00.007), and was gradually decreased among the without DR, non-proliferative DR and proliferative DR subgroups (P\xa0=\xa00.045). A logistic regression analysis showed that after adjusting for the other confounding factors, serum KL level was independently and negatively related with DR (P\xa0=\xa00.049). Furthermore, the increased apoptosis rates induced by PA were inhibited with the addition of KL protein. Consistently, KL protein reversed the expression levels of the increased pro-apoptotic protein Bax and the decreased anti-apoptotic protein Bcl-2 induced by PA. However, the anti-apoptotic effect of KL protein was attenuated by LY294002 through the phosphatidylinositol 3 kinase-serine∕threonine kinase pathway.The data suggested that KL protein was probably a potential protective factor against retinopathy in type\xa02 patients.© 2019 The Authors. Journal of Investigation published by Asian Association for the Study of (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: diabetes

High Fat Diet Upregulates Fatty Oxidation and Ketogenesis via Intervention of PPAR-γ.

Systemic hyperlipidemia and intracellular lipid accumulation induced by chronic high fat diet (HFD) leads to enhanced fatty oxidation (FAO) and ketogenesis. The present study was aimed to determine whether activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) by surplus free fatty acids (FA) in hyperlipidemic condition, has a positive feedback regulation over FAO and ketogenic enzymes controlling lipotoxicity and cardiac apoptosis.8 weeks old C57BL/6 wild type (WT) or PPAR-γ-/- mice were challenged with 16 weeks 60% HFD to induce obesity mediated type 2 (T2DM) and diabetic cardiomyopathy. Treatment course was followed by echocardiographic measurements, glycemic and lipid profiling, immunoblot, qPCR and immunohistochemistry (IHC) analysis of PPAR-γ and following mitochondrial metabolic enzymes 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), mitochondrial β- hydroxy butyrate dehydrogenase (BDH1) and pyruvate dehydrogenase kinase isoform 4 (PDK4). In vivo model was translated in vitro, with neonatal rat cardiomyocytes (NRCM) treated with PPAR-γ agonist/antagonist and PPAR-γ overexpression adenovirus in presence of (PA). Apoptosis was determined in vivo from left ventricular heart by TUNEL assay and immunoblot analysis.We found exaggerated circulating ketone bodies production and expressions of the related mitochondrial enzymes HMGCS2, BDH1 and PDK4 in HFD-induced diabetic hearts and in PA-treated NRCM. As a mechanistic approach we found HFD mediated activation of PPAR-γ is associated with the above-mentioned mitochondrial enzymes. HFD-fed PPAR-γ-/-mice display decreased hyperglycemia, hyperlipidemia associated with increased insulin responsiveness as compared to HFD-fed WT mice PPAR-γ-/-HFD mice demonstrated a more robust functional recovery after induction, as well as significantly reduced myocyte apoptosis and improved cardiac function.PPAR-γ has been described previously to regulate lipid metabolism and adipogenesis. The present study suggests for the first time that increased PPAR-γ expression by HFD is responsible for cardiac dysfunction via upregulation of mitochondrial enzymes HMGCS2, BDH1 and PDK4. Targeting PPAR-γ and its downstream mitochondrial enzymes will provide novel strategies in preventing metabolic and myocardial dysfunction in .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic metabolism-mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 .Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Metabolic flexibility to lipid availability during exercise is enhanced in individuals with high insulin sensitivity.

Metabolic flexibility to lipid (MetFlex-lip) is the capacity to adapt lipid oxidation to lipid availability. Hypothetically, impaired MetFlex-lip in skeletal muscle induces accumulation of lipid metabolites that interfere with insulin signaling. Our aim was to compare MetFlex-lip during exercise in subjects with low (Low_IS) vs. high (High_IS) insulin sensitivity. Twenty healthy men were designated as Low_IS or High_IS on the basis of the median of the homeostatic model assessment of insulin resistance index. Groups had similar age, body mass index, and maximum oxygen uptake (V̇o). Subjects cycled at 50% V̇o until expending 650 kcal. Adaptation in lipid oxidation was calculated as the drop in respiratory quotient (RQ) at the end of exercise vs. the maximum RQ (ΔRQ). Lipid availability was calculated as the increase in circulating nonesterified fatty acids (NEFA) at the end of exercise vs. the minimum NEFA (ΔNEFA). ΔRQ as a function of ΔNEFA was used to determine MetFlex-lip. On average, RQ and circulating NEFA changed similarly in both groups. However, ΔRQ correlated with ΔNEFA in High_IS ( r\u2009=\u2009-0.83, P < 0.01) but not in Low_IS ( r\u2009=\u2009-0.25, P = 0.48) subjects. Thus the slope of the ΔRQ vs. ΔNEFA relationship was steeper in High_IS vs. Low_IS subjects (-0.139 ± 0.03 vs. -0.025 ± 0.03 RQ·mmol·l, respectively; P < 0.05), with similar intercepts. We conclude that in subjects with High_IS lipid-to-carbohydrate oxidation ratio adapts to the increased circulating NEFA availability during exercise. Such MetFlex-lip appears impaired in subjects with Low_IS. Whether a cause-effect relationship exists between impaired MetFlex-lip and low insulin sensitivity remains to be determined.

Keyword: diabetes

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for obesity and .© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

Saturated fatty stimulates production of extracellular vesicles by renal tubular epithelial cells.

Lipotoxicity, an accumulation of intracellular lipid metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of metabolic disease. , a predominant lipid derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated (PA), and BSA-conjugated oleic (OA) for 24-48\xa0h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30-150\xa0nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of lipid-induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of lipid-induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keyword: diabetes

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced insulin resistance that defines in part the mechanisms of type 2 and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty metabolism, insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

Inhibition of insulin resistance by PGE1 via autophagy-dependent FGF21 pathway in diabetic nephropathy.

Insulin resistance is a critical process in the initiation and progression of diabetic nephropathy (DN). Alprostadil (Prostaglandin E1, PGE1) had protective effects on renal function. However, it is unknown whether PGE1 inhibited insulin resistance in renal tubule epithelial cells via autophagy, which plays a protective role in DN against insulin resistance. Insulin resistance was induced by (PA) in human HK-2 cells, shown as the decrease of insulin-stimulated AKT phosphorylation, glucose transporter-4 (GLUT4), glucose uptake and enhanced phosphorylation of insulin receptor substrate 1(IRS-1) at site serine 307 (pIRS-1ser307) and downregulated expression of IRS-1. Along with less abundance of p62, autophagy markers LC3B and Beclin-1 significantly increased in HK-2 cells exposed to PA. Such abnormal changes were significantly reversed by PGE1, which mimicked the role of autophagy gene 7 small interfering RNA (ATG7 siRNA). Furthermore, PGE1 promoted the protein expression of autophagy-related fibroblast growth factor-21 (FGF21), which alleviated insulin resistance. Results from western blotting and immunohistochemistry indicated that PGE1 remarkably restored autophagy, insulin resistance and the FGF21 expression in rat kidney of type 2 (T2DM). Collectively, we demonstrated the potential protection of PGE1 on insulin resistance in renal tubules via autophagy-dependent FGF21 pathway in preventing the progression of DN.

Keyword: diabetes

Palmitate and Stearate are Increased in the Plasma in a 6-OHDA Model of Parkinson\'s Disease.

Parkinson\'s disease (PD) is the second most common neurodegenerative disorder, without any widely available curative therapy. Metabolomics is a powerful tool which can be used to identify unexpected pathway-related disease progression and pathophysiological mechanisms. In this study, metabolomics in brain, plasma and liver was investigated in an experimental PD model, to discover small molecules that are associated with dopaminergic cell loss.Sprague Dawley (SD) rats were injected unilaterally with 6-hydroxydopamine (6-OHDA) or saline for the vehicle control group into the medial forebrain bundle (MFB) to induce loss of dopaminergic neurons in the substantia nigra pars compacta. Plasma, midbrain and liver samples were collected for metabolic profiling. Multivariate and univariate analyses revealed metabolites that were altered in the PD group.In plasma, ( = 3.72 × 10, FC = 1.81) and stearic ( = 3.84 × 10, FC = 2.15), were found to be increased in the PD group. ( = 3.5 × 10) and stearic ( = 2.7 × 10) correlated with test scores indicative of motor dysfunction. Monopalmitin ( = 4.8 × 10, FC = -11.7), monostearin ( = 3.72 × 10, FC = -15.1) and myo-inositol ( = 3.81 × 10, FC = -3.32), were reduced in the midbrain. The liver did not have altered levels of these molecules.Our results show that saturated free fatty acids, their monoglycerides and myo-inositol metabolism in the midbrain and enteric circulation are associated with 6-OHDA-induced PD pathology.

Keyword: diabetes

Sodium Orthovanadate Changes Fatty Composition and Increased Expression of Stearoyl-Coenzyme A Desaturase in THP-1 Macrophages.

Vanadium compounds are promising antidiabetic agents. In addition to regulating glucose metabolism, they also alter lipid metabolism. Due to the clear association between and atherosclerosis, the purpose of the present study was to assess the effect of sodium orthovanadate on the amount of individual fatty acids and the expression of stearoyl-coenzyme A desaturase (SCD or Δ-desaturase), Δ-desaturase, and Δ-desaturase in macrophages. THP-1 macrophages differentiated with phorbol 12-myristate 13-acetate (PMA) were incubated in vitro for 48\xa0h with 1\xa0μM or 10\xa0μM sodium orthovanadate (NaVO). The estimation of fatty composition was performed by gas chromatography. Expressions of the genes SCD, fatty desaturase 1 (FADS1), and fatty desaturase 2 (FADS2) were tested by qRT-PCR. Sodium orthovanadate in THP-1 macrophages increased the amount of saturated fatty acids (SFA) such as and stearic , as well as monounsaturated fatty acids (MUFA)-oleic and palmitoleic . Sodium orthovanadate caused an upregulation of SCD expression. Sodium orthovanadate at the given concentrations did not affect the amount of polyunsaturated fatty acids (PUFA) such as linoleic , arachidonic , eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA). In conclusion, sodium orthovanadate changed SFA and MUFA composition in THP-1 macrophages and increased expression of SCD. Sodium orthovanadate did not affect the amount of any PUFA. This was associated with a lack of influence on the expression of FADS1 and FADS2.

Keyword: diabetes

Docosahexaenoic antagonizes the boosting effect of on LPS inflammatory signaling by inhibiting gene transcription and ceramide synthesis.

It is well known that saturated fatty acids (SFAs) and unsaturated fatty , in particular omega-3 polyunsaturated fatty acids (n-3 PUFAs), have different effects on inflammatory signaling: SFAs are pro-inflammatory but n-3 PUFAs have strong anti-inflammatory properties. We have reported that (PA), a saturated fatty , robustly amplifies lipopolysaccharide (LPS) signaling to upregulate proinflammatory gene expression in macrophages. We also reported that the increased production of ceramide (CER) via sphingomyelin (SM) hydrolysis and CER de novo synthesis plays a key role in the synergistic effect of LPS and PA on proinflammatory gene expression. However, it remains unclear if n-3 PUFAs are capable of antagonizing the synergistic effect of LPS and PA on gene expression and CER production. In this study, we employed the above macrophage culture system and lipidomical analysis to assess the effect of n-3 PUFAs on proinflammatory gene expression and CER production stimulated by LPS and PA. Results showed that DHA strongly inhibited the synergistic effect of LPS and PA on proinflammatory gene expression by targeting nuclear factor kappa B (NFκB)-dependent gene transcription. Results also showed that DHA inhibited the cooperative effect of LPS and PA on CER production by targeting CER de novo synthesis, but not SM hydrolysis. Furthermore, results showed that myriocin, a specific inhibitor of serine palmitoyltransferase, strongly inhibited both LPS-PA-stimulated CER synthesis and proinflammatory gene expression, indicating that CER synthesis is associated with proinflammatory gene expression and that inhibition of CER synthesis contributes to DHA-inhibited proinflammatory gene expression. Taken together, this study demonstrates that DHA antagonizes the boosting effect of PA on LPS signaling on proinflammatory gene expression by targeting both NFκB-dependent transcription and CER de novo synthesis in macrophages.

Keyword: diabetes

Salvianolic B Attenuates Apoptosis of HUVEC Cells Treated with High Glucose or High Fat via Sirt1 Activation.

High glucose and high fat are important inducements for the development and progression of diabetic cardiopathy. Salvianolic B (SAB), which is the most abundant and bioactive compound in Danshen, attenuates oxidative stress-related disorders, such as cardiovascular diseases, cerebral ischemia, and . However, the effect of SAB on diabetic cardiopathy is not clear. The aim of study was to investigate the effect and the underlying molecular mechanisms of SAB on diabetic cardiopathy in vitro model. The human umbilical vein endothelial (HUVEC) cells were treated with high glucose (HG, 30 mM) or high fat (, PA, 0.75 mM) in the presence or absence of SAB (100, 200, and 400 mg/L) and incubated for 24 h. We found that HG or PA induced apoptosis of HUVEC cells, while treatment with SAB inhibited the apoptosis. We also found that SAB reversed HG- or PA-induced oxidative stress, apoptosis cell cytokines production, and expression of thioredoxin-interacting protein (TXNIP). Moreover, SAB increased HG- or PA-induced expression of Sirtuin 1 (Sirt1), a nicotinamide adenine dinucleotide- (NAD-) dependent histone deacetylase. Exposure of HUVEC cells to Ex527 (Sirt1 inhibitor) suppressed the effect of SAB on acetyl-p53 and procaspase-3 expressions. In conclusion, the results suggested that SAB could attenuate HUVEC cells damage treated with HG or PA via Sirt1 and might be a potential therapy agent for the diabetic cardiopathy treatment.

Keyword: diabetes

Plasma fatty acids in de novo lipogenesis pathway are associated with diabetogenic indicators among adults: NHANES 2003-2004.

Insulin regulates fatty acids (FAs) in the blood; conversely, FAs may mediate insulin sensitivity and are potentially modifiable risk factors of the diabetogenic state.The objective of our study was to examine the associations between plasma concentrations of FAs, fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among individuals (n\xa0=\xa01433) in the NHANES (2003-2004).Plasma concentrations of 24 individual FAs were considered individually and in subgroups, per chemical structure. Study participants were categorized in diabetogenic groups: Group 1 (HbA1c ≥6.5% or FPG ≥126 mg/dL), Group 2 (HbA1c 5.7% to <6.5% or FPG 100 to <126 mg/dL), and Group 3 (HbA1c <5.7% and FPG <100 mg/dL). We assessed associations between diabetogenic groups and plasma FAs in multivariate multinomial regressions (with Group 3 as the reference).Overall, 7.0% of study participants were in Group 1; 33.3% were in Group 2. Plasma concentrations of several individual FAs, including even-chain saturated FAs (SFAs; myristic, , stearic acids) and monounsaturated FAs (MUFAs; cis-vaccenic, oleic acids), were respectively associated with greater odds of Groups 1 and 2 status, adjusting for covariates. Higher concentrations of SFA and MUFA subgroups (highest compared with lowest quartile) were associated with increased odds of Group 2 status [SFAs adjusted OR (aOR): 1.51 (95% CI: 1.05, 2.18); MUFAs aOR: 1.78 (95% CI: 1.11, 2.85)]. Higher eicosapentaenoic plasma concentration was associated with decreased odds of Group 1 status [quartile 4 aOR: 0.41 (95% CI: 0.17, 0.95)].Higher plasma concentrations of SFAs and MUFAs, primary de novo lipogenesis products, were associated with elevated FPG and HbA1c in a nationally representative study population in the United States. Additional studies are necessary to elucidate potential causal relationships between FAs (from endogenous production and dietary consumption) and diabetogenic indicators, as well as clinical implications for managing and prediabetes.

Keyword: diabetes

Pterostilbene reverses mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: diabetes

and Oleic : The Yin and Yang of Fatty Acids in Type 2 .

Increased plasma non-esterified fatty acids (NEFAs) link obesity with insulin resistance and type 2 (T2DM). However, in contrast to the saturated FA (SFA) , the monounsaturated FA (MUFA) oleic elicits beneficial effects on insulin sensitivity, and the dietary :oleic ratio impacts risk in humans. Here we review recent mechanistic insights into the beneficial effects of oleic compared with on insulin resistance and T2DM, including its anti-inflammatory actions, and its capacity to inhibit endoplasmic reticulum (ER) stress, prevent attenuation of the insulin signaling pathway, and improve β cell survival. Understanding the molecular mechanisms of the antidiabetic effects of oleic may contribute to understanding the benefits of this FA in the prevention or delay of T2DM.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Circulating Linoleic is Associated with Improved Glucose Tolerance in Women after Gestational .

Women with previously diagnosed gestational (GDM) are at increased risk of type-2- (T2D). We aimed to establish links between glucose tolerance (GT) and serum fatty (FA) profile in the transition from GDM to T2D. Six years after GDM, 221 women were grouped as having normal GT (NGT), impaired GT (IGT), or T2D based on oral GT test results. Fasting serum FAs were profiled, anthropometric measures taken, and dietary intake determined. Linoleic (LA) was significantly higher in NGT women ( < 0.001) compared with IGT and T2D, and emerged as a strong predictor of low glucose and insulin levels, independently of BMI. Self-reported vegetable oil consumption correlated with LA serum levels and glucose levels. Delta-6-, delta-9-, and stearoyl-CoA-desaturase activities were associated with decreased GT, and delta-5-desaturase activities with increased GT. In a subgroup of women at high risk of , low LA and high levels were seen in those that developed T2D, with no differences in other FAs or metabolic measurements. Results suggest that proportions of LA and are of particular interest in the transition from GDM to T2D. Interconversions between individual FAs regulated by desaturases appear to be relevant to glucose metabolism.

Keyword: diabetes

Argirein alleviates vascular endothelial insulin resistance through suppressing the activation of Nox4-dependent O production in diabetic rats.

Insulin resistance in endothelial cells contributes to the development of cardiovascular disease in type 2 (T2DM). Therefore, there are great potential clinical implications in developing pharmacological interventions targeting endothelial insulin resistance. Our previous studies indicated that argirein which was developed by combining rhein with L-arginine by a hydrogen bond, could substantially relieved stress related exacerbation of cardiac failure and alleviated cardiac dysfunction in T2DM, which was associated with suppressing NADPH oxidase activity. However, it is unclear whether argirein treatment attenuates the vascular lesion and dysfunction in T2DM and its underlying mechanisms.The rat aortic endothelial cells (RAECs) were used to treat with (PA), a most common saturated free fatty , which could induce insulin resistance. It was showed that argirein increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of IRS-1-ser307 and AKT-ser473, consequently resulting in the increase of the production of eNOS and NO in PA-induced RAECs. We further found that argirein blocked the Nox4-dependent superoxide (O) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vitro, argirein increased the release of endothelial NO to relieve the vasodilatory response to acetylcholine and insulin, and restored the expression of Nox4 and pIRS-1-ser307 in the aorta endothelium of high-fat diet (HFD)-fed rats following an injection of streptozocin (STZ).These results suggested that argirein could improve endothelial insulin resistance which was attributed to inhibiting Nox4-dependent redox signaling in RAECs. These studies thus revealed the novel effect of argirein to prevent the vascular complication in T2DM.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of , hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: diabetes

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty metabolism was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. Lipid distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral lipid metabolic process" and "Regulation of lipid storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body energy metabolism.

Keyword: diabetes

Blood Fatty Profiles: New Biomarkers for Cardiometabolic Disease Risk.

Fatty (FA) profiles in different blood compartments are reflections of both diet and metabolism, and some FA levels are related to disease risk.Perhaps the most studied FA-disease relationship is between long-chain omega-3 polyunsaturated fatty acids [eicosapentaenoic (EPA) and docosahexaenoic (DHA)] and cardiovascular disease (CVD). Despite null results from recent large omega-3 FA supplementation trials, new research continues to support past studies showing that blood levels of EPA\u2009+\u2009DHA are inversely related to risk for total mortality and fatal CVD events. But blood levels of other FAs may also be useful markers of risk for a variety of diseases. The essential omega-6 FA linoleic is inversely associated with risk for developing type 2 (T2D), whereas risk for T2D is directly related to biomarkers of de novo lipogenesis ( and palmitoleic acids). Levels of industrially produced trans FAs have been linked to higher risk for CVD. Thus, blood levels of several individual FAs are emerging as modifiable biomarkers for risk of major chronic diseases.

Keyword: diabetes

-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and Inflammation via Nuclear Mechanisms.

Objective- Macrophages play key roles in inflammation and diabetic vascular complications. Emerging evidence implicates long noncoding RNAs in inflammation, but their role in macrophage dysfunction associated with inflammatory diabetic complications is unclear and was therefore investigated in this study. Approach and Results- RNA-sequencing and real-time quantitative PCR demonstrated that a long noncoding RNA Dnm3os (dynamin 3 opposite strand) is upregulated in bone marrow-derived macrophages from type 2 diabetic db/db mice, diet-induced insulin-resistant mice, and diabetic ApoE mice, as well as in monocytes from type 2 diabetic patients relative to controls. Diabetic conditions (high glucose and ) induced Dnm3os in mouse and human macrophages. Promoter reporter analysis and chromatin immunoprecipitation assays demonstrated that diabetic conditions induce Dnm3os via NF-κB activation. RNA fluorescence in situ hybridization and real-time quantitative PCRs of subcellular fractions demonstrated nuclear localization and chromatin enrichment of Dnm3os in macrophages. Stable overexpression of Dnm3os in macrophages altered global histone modifications and upregulated inflammation and immune response genes and phagocytosis. Conversely, RNAi-mediated knockdown of Dnm3os attenuated these responses. RNA pull-down assays with macrophage nuclear lysates identified nucleolin and ILF-2 (interleukin enhancer-binding factor 2) as protein binding partners of Dnm3os, which was further confirmed by RNA fluorescence in situ hybridization immunofluorescence. Furthermore, nucleolin levels were decreased in diabetic conditions, and its knockdown enhanced Dnm3os-induced inflammatory gene expression and histone H3K9-acetylation at their promoters. Conclusions- These results demonstrate novel mechanisms involving upregulation of long noncoding RNA Dnm3os, disruption of its interaction with nucleolin, and epigenetic modifications at target genes that promote macrophage inflammatory phenotype in . The data could lead to long noncoding RNA-based therapies for inflammatory complications.

Keyword: diabetes

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in . Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: diabetes

Downregulation of miR-320 Alleviates Endoplasmic Reticulum Stress and Inflammatory Response in 3T3-L1 Adipocytes.

MicroRNAs serve important roles in the regulation of endoplasmic reticulum stress (ERs). This study aimed to investigate the role of microRNA-320 (miR-320) in the development of ERs and the inflammatory response in 3T3-L1 adipocytes.The adipose tissue expression levels of miR-320 and ERs markers (GRP78, GRP94, Derlin-1 and CHOP) and the serum concentration of inflammatory cytokines (TNF-α, NF-κB and IL-6) in obese patients were evaluated using quantitative real-time RT-PCR. The correlation of miR-320 with genes involved in ERs and inflammation was analyzed. The effects of miR-320 on ERs and inflammation were explored using mature 3T3-L1 adipocytes, which were pretreated with (PA).ERs markers and inflammatory cytokines were all upregulated in obese patients. Adipose tissue miR-320 expression was also increased in obese patients, and had positive correlations with the levels of ERs markers and inflammatory cytokines. After PA treatment, the levels of ERs markers and inflammatory cytokines were elevated significantly in 3T3-L1 adipocytes. Moreover, miR-320 expression was increased in the cells under ERs status. The upregulation of miR-320 could enhance the expression of ERs markers and inflammatory cytokines, but the downregulation of miR-320 resulted in the opposite results.The data of this study indicate that miR-320 expression is upregulated in ERs status, and the downregulation of miR-320 ameliorates ERs and the inflammatory response in 3T3-L1 adipocytes. We consider that the approaches to decrease miR-320 expression may be novel therapeutic strategies for the treatment of obesity and obesity-related diseases.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

Type 2 Promotes Cell Centrosome Amplification via AKT-ROS-Dependent Signalling of ROCK1 and 14-3-3σ.

Type 2 is associated with oxidative stress and DNA damage which can cause centrosome amplification. Thus, the study investigated centrosome amplification in type 2 and the underlying mechanisms.Centrosome numbers in human peripheral blood mononuclear blood cells (PBMC) from healthy subjects and patients with type 2 were compared to access the association between type 2 and centrosome amplification. Colon cancer cells were used to investigate the molecular mechanisms underlying the centrosome amplification triggered by high glucose, insulin and . Western blot analysis was used to quantify the level of protein and protein phosphorylation. Immunofluorescent staining was performed to detect centrosomes. ROS was quantified using flow cytometry technique. Transcriptpmic profiling was performed using Illumina HiSeqTM500 platform.We found that centrosome amplification was increased PBMC from the type 2 diabetic patients, which correlated with the levels of fasting blood glucose and HbA1c. High glucose, insulin and , alone or in combinations, induced ROS production and centrosome amplification. Together, they increased AKT activation as well as the expression, binding and centrosome translation of ROCK1 and 14-3-3σ. Results from further analyses showed that AKT-ROS-dependent upregulations of expression, binding and centrosome translocation of ROCK1 and 14-3-3σ was the molecular pathway underlying the centrosome amplification in vitro triggered by high glucose, insulin and . Moreover, the key in vitro molecular signalling events activated by high glucose, insulin and were verified in PBMC from the patients with type 2 .Our results show that type 2 promotes cell centrosome amplification, and suggest that the diabetic pathophysiological factors-activated AKT-ROS-dependent signalling of ROCK1 and 14-3-3σ is the underlying molecular mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Role of glutamine synthetase in angiogenesis beyond glutamine synthesis.

Glutamine synthetase, encoded by the gene GLUL, is an enzyme that converts glutamate and ammonia to glutamine. It is expressed by endothelial cells, but surprisingly shows negligible glutamine-synthesizing activity\xa0in these cells at physiological glutamine levels. Here we show in mice that genetic deletion of Glul in endothelial cells impairs vessel sprouting during vascular development, whereas pharmacological blockade of glutamine synthetase suppresses angiogenesis in ocular and inflammatory skin disease while only minimally affecting healthy adult quiescent endothelial cells. This relies on the inhibition of endothelial cell migration but not proliferation. Mechanistically we show that in human umbilical vein endothelial cells GLUL knockdown reduces membrane localization and activation of the GTPase RHOJ while activating other Rho GTPases and Rho kinase, thereby inducing actin stress fibres and impeding endothelial cell motility. Inhibition of Rho kinase rescues the defect in endothelial cell migration that is induced by GLUL knockdown. Notably, glutamine synthetase palmitoylates itself and interacts with RHOJ to sustain RHOJ palmitoylation, membrane localization and activation. These findings reveal that, in addition to the\xa0known formation of glutamine, the enzyme glutamine synthetase shows unknown\xa0activity in endothelial cell migration during pathological angiogenesis\xa0through RHOJ palmitoylation.

Keyword: diabetes

Rare ginsenosides ameliorate lipid overload-induced myocardial insulin resistance via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 . However, their pharmacological studies regarding myocardial fuel metabolism and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel metabolism and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high fat diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of lipid overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: diabetes

Metabolomics analysis of the protective effect of rubusoside on -induced lipotoxicity in INS-1 cells using UPLC-Q/TOF MS.

is one of the most severe chronic diseases worldwide. It is widely accepted that apoptosis of the pancreatic beta cell is an important cause for the induction of hyperglycemia and high levels of free fatty acids (FFAs), also called lipotoxicity associated with pancreatic beta cell dysfunction. Lipotoxicity has been proven to be an important pathogenic factor of . However, until now, the mechanism of FFA-induced lipotoxicity in INS-1 cells has not been fully understood. Current anti-diabetic drugs that protect islet cells are often toxic to healthy cells, resulting in negative side effects. Thus, there is an urgent need to identify more effective and safer anti-diabetic agents to protect pancreatic islet cells. Rubusoside (RUB) is a major ingredient in the leaves of Rubus suavissimus S. Lee, which decreases blood glucose levels by protecting pancreatic islet cells. However, the exact mechanism of this effect is unknown. In this study, metabolomics experiments based on UPLC-Q/TOF MS characterized a total of 15 metabolites as potential biomarkers associated with lipotoxicity induced by in INS-1 cells. According to the metabolic pathway analysis, pentose and glucuronate interconversions, and glycerophospholipid metabolism were recognized as the most influenced metabolic pathways associated with lipotoxicity. Unexpectedly, deviations of 14 metabolites in lipotoxic INS-1 cells were regulated by RUB, suggesting synergistic mediation of the abnormal metabolic pathways. The metabolomics strategy based on UPLC/Q-TOF MS analysis provides a new insight into the mechanisms of lipotoxicity induced by and the anti-lipotoxic activity of RUB in INS-1 cells.

Keyword: diabetes

The role and mechanism of K3.1 channels in human monocyte migration induced by .

Monocyte migration into diseased tissues contributes to the pathogenesis of diseases. Intermediate-conductance Ca-activated K (K3.1) channels play an important role in cell migration. However, the role of K3.1 channels in mediating monocyte migration induced by (PA) is still unclear. Using cultured THP-1 cells and peripheral blood mononuclear cells from healthy subjects, we investigated the role and signaling mechanisms of K3.1 channels in mediating the migration induced by PA. Using methods of Western blotting analysis, RNA interference, cell migration assay and ELISA, we found that PA-treated monocytes exhibited increment of the protein levels of K3.1 channel and monocyte chemoattractant protein-1 (MCP-1), and the effects were reversed by co-incubation of PA with anti-TLR2/4 antibodies or by specific inhibitors of p38-MAPK, or NF-κB. In addition, PA increased monocyte migration, which was abolished by a specific K3.1 channel blocker, TRAM-34, or K3.1 small interfering RNA (siRNA). The expression and secretion of MCP-1 induced by PA was also similarly prevented by TRAM-34 and K3.1 siRNA. These results demonstrate for the first time that PA upregulates K3.1 channels through TLR2/4, p38-MAPK and NF-κB pathway to promote the expression of MCP-1, and then induce the trans-endothelial migration of monocytes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 , which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture.

Previous work showed that muscle-derived stem cells (MDSCs) exposed long-term to the milieu of uncontrolled type 2 (UC-T2D) in male obese Zucker (OZ) rats, were unable to correct the associated erectile dysfunction and the underlying histopathology when implanted into the corpora cavernosa, and were also imprinted with a noxious gene global transcriptional signature (gene-GTS), suggesting that this may interfere with their use as autografts in stem cell therapy.To ascertain the respective contributions of dyslipidemia and hyperglycemia to this MDSC damage, clarify its mechanism, and design a bioassay to identify the damaged stem cells.Early MDSCs and late MDSCs were respectively isolated from nearly normal young OZ rats and moderately hyperglycemic and severely dyslipidemic/obese aged rats with erectile dysfunction. Monolayer cultures of early diabetic MDSCs were incubated 4 days in DMEM/10% fetal calf serum\xa0+ or\xa0- aged OZ or lean Zucker serum from non-diabetic lean Zucker rats (0.5-5%) or with soluble (PA) (0.5-2 mM), cholesterol (CHOL) (50-400 mg/dL), or glucose (10-25 mM).Fat infiltration was estimated by Oil red O, apoptosis by TUNEL, protein expression by Western blots, and gene-GTS and microRNA (miR)-GTS were determined in these stem cells\' RNA.Aged OZ serum caused fat infiltration, apoptosis, myostatin overexpression, and impaired differentiation. Some of these changes, and also a proliferation decrease occurred with PA and CHOL. The gene-GTS changes by OZ serum did not resemble the in\xa0vivo changes, but some occurred with PA and CHOL. The miR-GTS changes by OZ serum, PA, and CHOL resembled most of the in\xa0vivo changes. Hyperglycemia did not replicate most alterations.MDSCs may be damaged in long-term UC-T2D/obese patients and be ineffective in autologous human stem cell therapy, which may be prevented by excluding the damaged MDSCs.The in\xa0vitro test of MDSCs is innovative and fast to define dyslipidemic factors inducing stem cell damage, its mechanism, prevention, and counteraction. Confirmation is required in other T2D/obesity rat models and stem cells (including human), as well as miR-GTS biomarker validation as a stem cell damage biomarker.Serum from long-term UC-T2D/obese rats or dyslipidemic factors induces a noxious phenotype and miR-GTS on normal MDSCs, which may lead in\xa0vivo to the repair inefficacy of late diabetic MDSCs. This suggests that autograft therapy with MDSCs in long-term UT-T2D obese patients may be ineffective, albeit this may be predictable by prior stem cell miR-GTS tests. Masouminia M, Gelfand R, Kovanecz I, et\xa0al. Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture. J Sex Med 2018;15:1678-1697.Copyright © 2018 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

A prospective and longitudinal study of plasma phospholipid saturated fatty profile in relation to cardiometabolic biomarkers and the risk of gestational .

Data on saturated fatty acids (SFAs) in relation to metabolic function and glucose homeostasis remain controversial. Such data are lacking among pregnant women.We prospectively investigated objectively measured individual and subclasses of plasma phospholipid SFAs throughout pregnancy in relation to cardiometabolic markers and gestational (GDM) risk.Within the National Institute of Child Health and Human Development Fetal Growth Studies-Singleton Cohort of 2802 singleton pregnancies, 107 GDM cases were ascertained via medical record review and matched to 214 non-GDM controls on age, race/ethnicity, and gestational week (GW) at blood collection. Individual plasma phospholipid SFA concentrations were repeatedly measured throughout pregnancy at GWs 10-14, 15-26, 23-31, and 33-39 and also grouped into subclasses of even- or odd-chain SFAs.From GW 10, even-chain SFA concentrations were significantly higher among women who later developed GDM, whereas odd-chain SFAs were significantly lower among GDM cases compared with controls. At GWs 10-14, the SFA (16:0) was positively associated with impaired insulin resistance and cardiometabolic markers and the risk of GDM [adjusted OR comparing the highest with the lowest quartile (aORQ4-Q1): 4.76; 95% CI: 1.72, 13.10; P-trend\xa0=\xa00.001]. In contrast, odd-chain SFAs were inversely related to the previously mentioned markers and GDM risk [aORQ4-Q1 for pentadecanoic (15:0): 0.32; 95% CI: 0.11, 0.92; P-trend\xa0=\xa00.025; for heptadecanoic (17:0): 0.20; 95% CI: 0.07, 0.58; P-trend\xa0=\xa00.003]. Women with high (median or greater) even-chain SFA concentrations and low (less than median) odd-chain SFAs had a 9.43-fold (95%: CI 3.26-, 27.30-fold) increased risk compared with women with low even-chain and high odd-chain SFA concentrations. Similar results were observed at GWs 15-26.The study provided one of the first lines of evidence suggesting that circulating concentrations of SFAs varying by SFA chain length, as early as GWs 10-14, were significantly and differentially associated with subsequent risk of GDM. Our findings highlight the importance of assessing objectively measured, individual, and subclasses of SFAs to investigate their distinct biological and pathophysiologic roles in glucose homeostasis and cardiometabolic outcomes. This study was registered at www.clinicaltrials.gov as .

Keyword: diabetes

Sirtuin 5 overexpression attenuates glucolipotoxicity-induced pancreatic β cells apoptosis and dysfunction.

Recently, SIRT5 was reported to be a predominant desuccinylase and demalonylase in mitochondria. Ablation of SIRT5 enhances the systemic succinylation and malonylation of mitochondrial proteins, including various metabolic enzymes; however, its function in pancreatic β cells has not yet been clarified. In this study, we evaluated the effects of SIRT5 overexpression on glucolipotoxicity-induced apoptosis in β cell lines. Full-length SIRT5, which preferentially targeted to mitochondria and partially to the nucleus and cytoplasm, was overexpressed in NIT-1 cells. Chronic exposure to excess palmitate and glucose (High-PA-G) induced apoptosis and suppressed glucose-stimulated insulin secretion in β cells. SIRT5 overexpression significantly alleviated apoptosis under the High-PA-G condition, accompanied by suppressed Caspase 3 activity and reduced malondialdehyde levels. SIRT5 overexpression also improved β cell secretory capacity in response to glucose under the High-PA-G condition, suggesting its protective role in β cell function. Furthermore, SIRT5 overexpression reversed the decreasing trend of anti-apoptotic factors BCL-2 and BCL-XL expression under High-PA-G condition. Further regulation mechanisms between SIRT5 and these anti-apoptotic factors remains to be explored in future studies. Our data reveal that SIRT5 is a potentially protective factor for pancreatic β cells against glucolipotoxicity-induced apoptosis and cell dysfunction.Copyright © 2018. Published by Elsevier Inc.

Keyword: diabetes

Postprandial Saturated Fatty Acids Increase the Risk of Type 2 : A Cohort Study in a Chinese Population.

Experimental evidence suggests saturated fatty acids (SFAs) are associated with insulin resistance, but results from epidemiological studies on fasting SFAs- risk are inconsistent.We investigated SFA (fasting and 2-hour postprandial) profiles and risk.A total of 8940 participants were recruited for the Harbin People\'s Health Study in 2008. Serum SFAs (fasting and 2-hour postprandial) at baseline in Chinese men and women without were profiled, and type 2 was ascertained using World Health Organization criteria after 4 to 7 years of follow-up.Associations between 2-hour postprandial SFA (2h-SFA) and .At baseline, incident cases of were older with a higher body mass index and waist circumference. After a mean follow-up of 6.7 years, 658 incident cases of occurred. After propensity score computation and inverse probability of treatment weighting (IPTW) estimation, fasting SFAs were unrelated to risk but IPTW-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the highest tertile of 2-hour postprandial stearic (2h-SA), 2-hour postprandial (2h-PA), and 2h-SFA for risk were 2.50 (2.08 to 3.16), 1.56 (1.23 to 2.02), and 1.70 (1.34 to 2.17), respectively (P-trend < 0.0001). Similarly, 2h-SA/fasting SA, 2h-PA/fasting PA, and 2h-SFA/fasting SFA ratios [IPTW-adjusted OR (95% CI): 2.94 (2.39 to 3.58), 2.31 (1.80 to 2.93), and 2.42 (1.91 to 3.11), respectively; P-trend < 0.0001] predicted the risk.Higher serum 2h-SFA (but not fasting SFA) independently predicted risk.

Keyword: diabetes

Changes of MODY signal pathway genes in the endoplasmic reticulum stress in INS-1-3 cells.

Metabolic disturbances induce endoplasmic reticulum stress (ERS) in pancreatic beta cells. This study aims to investigate whether a common pathway exists in the ERS induced by various chemicals, including high levels of glucose and palmitate in INS-1-3 cells.ERS in INS-1-3 cells was induced by exposure cells to thapsigargin (TG), tunicamycin (TM) or (PA) +high glucose (HG). Digital gene expression (DGE) profiling technique was used to detect differentially expressed genes. The profile of gene expression was detected by gene oncology (GO) function and pathway enrichment analysis. Nkx6.1 over-expression was established in INS-1-3 cell lines by lentivirus infection to revert the inhibition of Nkx6.1 expression found in the situation of ERS. Real time reverse transcription polymerase chain reaction (RT-PCR) was used to verify the expression changes of key genes. Cell viability was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptosis was determined by flow cytometry. INS-1-3 cell function was measured by glucose stimulated insulin secretion test(GSIS).As compared to control, DGE demonstrated that there were 135, 57 and 74 differentially expressed genes in TM, TG and HG+PA groups, respectively. Those differentially expressed genes were enriched to ERS, antigen processing and presentation, protein export pathways, and interestingly, the maturity onset of the young (MODY) pathway. Nkx6.1 is one of common down-regulated gene in MODY signaling pathway among TM, TG and HG+PA groups. Over-expression of Nkx6.1 ameliorated glucolipotoxicity induced apoptosis rate by 45.4%, and increased proliferation by 40.9%. At the same time, GSIS increased by 1.82 folds.MODY pathway genes expression was changed in the state of ERS. Over-expression of Nkx6.1 protected the INS-1-3 cells from glucolipotoxicity.

Keyword: diabetes

Rational design of dimeric lipidated Xenopus glucagon-like peptide 1 analogues as long-acting antihyperglycaemic agents.

Dimerization is viewed as an effective means to enhance the binding affinity and therapeutic potency of peptides. Both dimerization and lipidation effectively prolong the half-life of peptides in\xa0vivo by increasing hydrodynamic size and facilitating physical interactions with serum albumin. Here, we report a novel method to discover long-acting glucagon-like peptide 1 (GLP-1) analogues by rational design based on Xenopus GLP-1 through a combined dimerization and lipidization strategy. On the basis of our previous structure analysis of Xenopus GLP-1, and a C-terminal Cys were firstly introduced into two Xenopus GLP-1 analogues (1 and 2), and the afforded 3 and 4 were further reacted with bis-maleimide amine to afford two dimeric lipidated Xenopus GLP-1 analogues (5 and 6). The in\xa0vitro and in\xa0vivo biological activities of 5 and 6 were significantly improved as compared with their monomers. Moreover, the selected compound 6 showed greater hypoglycemic and insulinotropic activities than liraglutide even when the dose of 6 was reduced to half in db/db mice. Pharmacokinetic test revealed that 6 had a ∼ 3-fold longer half-life than liraglutide in Kunming mice and SD rats, and the longer half-life of 6 led to excellent long-acting hypoglycemic effects as confirmed by two different pharmacological methods conducted on db/db mice. Finally, a 7 weeks chronic study conducted on db/db mice demonstrated the better therapeutic efficacies of 6 on glucose tolerance normalization, HbA1c reduction and pancreas islets protection than liraglutide. The present research showed that combined dimerization and lipidization is effective when applied to Xenopus GLP-1 analogue to develop novel GLP-1 analogue for the treatment of type 2 . In addition, the promising preclinical data of 6 suggested the therapeutic potential of 6 as a novel anti-diabetic agent.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Excessive Autophagy Activation and Increased Apoptosis Are Associated with -Induced Cardiomyocyte Insulin Resistance.

Diabetic cardiomyopathy (DCM) remains the major cause of death associated with . Researchers have demonstrated the importance of impaired cardiac insulin signaling in this process. Insulin resistance (IR) is an important predictor of DCM. Previous studies examining the dynamic changes in autophagy during IR have yielded inconsistent results. This study aimed to investigate the dynamic changes in autophagy and apoptosis in the rat H9c2 cardiomyocyte IR model. H9c2 cells were treated with 500\u2009M (PA) for 24 hours, resulting in the induction of IR. To examine autophagy, monodansylcadaverine staining, GFP-LC3 puncta confocal observation, and Western blot analysis of LC3I-to-LC3II conversion were used. Results of these studies showed that autophagic vesicles increased in numbers during the first 24 hours and then decreased by 36 hours after PA treatment. Western blot analysis showed that treatment of H9c2 cells with 500\u2009M PA for 24 hours decreased the expression of Atg12-Atg5, Atg16L1, Atg3, and PI3Kp85. Annexin V/PI flow cytometry revealed that PA exposure for 24 hours increased the rate of apoptosis. Together, this study demonstrates that PA induces IR in H9c2 cells and that this process is accompanied by excessive activation of autophagy and increases in apoptosis.

Keyword: diabetes

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism.A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of -induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate -induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid metabolism were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of lipid droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and lipid levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic lipid droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid metabolism dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid metabolism in T2DM, which can inhibit hepatic lipogenesis and gluconeogenesis and elevate glycogen synthesis by targeting STAT3.

Keyword: diabetes

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: diabetes

Impact of Brain Fatty Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and energy balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with fatty signaling. In humans circulating saturated fatty acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of fatty acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic fatty signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain fatty signaling and its contemporaneous effect on liver function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

Sonodynamic therapy inhibits palmitate-induced beta cell dysfunction via PINK1/Parkin-dependent mitophagy.

In type 2 (T2DM), the overload of glucose and lipids can promote oxidative stress and inflammatory responses and contribute to the failure of beta cells. However, therapies that can modulate the function of beta cells and thus prevent their failure have not been well explored. In this study, beta cell injury model was established with (PA) to simulate the lipotoxicity (high-fat diet) found in T2DM. Sonodynamic therapy (SDT), a novel physicochemical treatment, was applied to treat injured beta cells. We found that SDT had specific effects on mitochondria and induced transient large amount of mitochondrial reactive oxygen species (ROS) production in beta cells. SDT also improved the morphology and function of abnormal mitochondria, inhibited inflammatory response and reduced beta cell dysfunction. The improvement of mitochondria was mediated by PINK1/Parkin-dependent mitophagy. Additionally, SDT rescued the transcription of PINK1 mRNA which was blocked by PA treatment, thus providing abundant PINK1 for mitophagy. Moreover, SDT also increased insulin secretion from beta cells. The protective effects of SDT were abrogated when mitophagy was inhibited by cyclosporin A (CsA). In summary, SDT potently inhibits lipotoxicity-induced beta cell failure via PINK1/Parkin-dependent mitophagy, providing theoretical guidance for T2DM treatment in aspects of islet protection.

Keyword: diabetes

Theaflavins Improve Insulin Sensitivity through Regulating Mitochondrial Biosynthesis in -Induced HepG2 Cells.

Theaflavins, the characteristic and bioactive polyphenols in black tea, possess the potential improving effects on insulin resistance-associated metabolic abnormalities, including obesity and type 2 . However, the related molecular mechanisms are still unclear. In this research, we investigated the protective effects of theaflavins against insulin resistance in HepG2 cells induced by . Theaflavins significantly increased glucose uptake of insulin-resistant cells at noncytotoxic doses. This activity was mediated by upregulating the total and membrane bound glucose transporter 4 protein expressions, increasing the phosphor-Akt (Ser473) level, and decreasing the phosphorylation of IRS-1 at Ser307. Moreover, theaflavins were found to enhance the mitochondrial DNA copy number, down-regulate the PGC-1β mRNA level and increase the PRC mRNA expression. Mdivi-1, a selective mitochondrial division inhibitor, could attenuate TFs-induced promotion of glucose uptake in insulin-resistant HepG2 cells. Taken together, these results suggested that theaflavins could improve hepatocellular insulin resistance induced by free fatty acids, at least partly through promoting mitochondrial biogenesis. Theaflavins are promising functional food ingredients and medicines for improving insulin resistance-related disorders.

Keyword: diabetes

Nutritionally Derived Metabolic Cues Typical of the Obese Microenvironment Increase Cholesterol Efflux Capacity of Adipose Tissue Macrophages.

Cholesterol retention within plasma membranes of macrophages is associated with increased inflammatory signaling. Cholesterol efflux via the transporters ABCA1, ABCG1, and SR-BI to high-density lipoprotein (HDL) particles is a critical mechanism to maintain cellular cholesterol homeostasis. Little is known about the impact of the obese microenvironment on cholesterol efflux capacity (CEC) of macrophages. In this study, the CEC of obese-derived primary adipose-tissue macrophages (ATM) is evaluated and the in vivo microenvironment is modeled in vitro to determine mechanisms underlying modulated CEC.F4/80 ATM are labeled with H-cholesterol ex vivo, and CEC and ABCA1/ABCG1 protein levels are determined. Total, ABCA1-dependent, and ABCA1-independent CECs are determined in J774 macrophages polarized to M1 (LPS&IFNγ), M2 (IL-4&IL-13), or metabolic phenotypes (glucose, insulin, and ).Obese ATM exhibit enhanced CEC and ABCA1 and ABCG1 expression compared to lean ATM. In contrast, ABCA1-CEC is suppressed from M1 polarized macrophages compared to untreated in vitro, by activation of the JAK/STAT pathway. Incubation of macrophages in vitro in high glucose augments cAMP-induced ABCA1 protein expression and ABCA1-CEC.These novel findings demonstrate remarkable plasticity of macrophages to respond to their environment with specific modulation of ABCA1 depending on whether classical pro-inflammatory or metabolic cues predominate.© 2018 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: diabetes

Individual free fatty acids have unique associations with inflammatory biomarkers, insulin resistance and insulin secretion in healthy and gestational diabetic pregnant women.

We investigated the relationships of maternal circulating individual free fatty acids (FFA) with insulin resistance, insulin secretion and inflammatory biomarkers during mid-pregnancy.The data were drawn from a prospective cohort of generally healthy pregnant women (n=1368, African-American 36%, Hispanic 48%, Caucasian 16%) in Camden, NJ. We quantitatively determined 11 FFAs, seven cytokine/adipokine, homeostatic model assessment of insulin resistance (HOMA-IR) and C-peptide levels from the fasting blood samples that were collected at 16 weeks of gestation. Multivariate analyses were performed along with separate analyses for each individual FFA.High HOMA-IR (p<0.001) and C-peptide (p<0.0001) levels were positively associated with a twofold to fourfold increased risk for developing gestational (GDM). Negative relationships were found with specific FFAs (molecular percentage, palmitoleic, oleic, linolenic, myristic acids) and HOMA-IR and C-peptide levels (p<0.01\u2009to p<0.0001). In contrast, , stearic, arachidonic, dihomo-γ-linolenic (DGLA) and docosahexaenoic acids were positively associated with HOMA-IR and C-peptide (p<0.01\u2009to p<0.0001). The individual FFAs also predicted cytokine/adipokine levels. For example, women who had elevated DGLA (highest quartile) were twice as (adjusted OR 2.06, 95%\u2009CI 1.42 to 2.98) likely to have higher interleukin (IL)-8 (p<0.0001) levels. Conversely, women with high palmitoleic, oleic, and linolenic levels had reduced odds (≥2-fold, p<0.01\u2009to p<0.001) for having higher IL-8, IL-6 or tumor necrosis factor-alpha levels.Our results suggest that maternal individual FFAs uniquely affect insulin resistance and secretion. The effects are either direct or indirect via modulation of the inflammatory response. Modifying the composition of FFAs may help in reducing the risk of GDM.

Keyword: diabetes

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid metabolism-related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 . However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of lipid metabolism- and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid metabolism-related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid metabolism, suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in . In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: diabetes

MicroRNA‑29a is involved lipid metabolism dysfunction and insulin resistance in C2C12 myotubes by targeting PPARδ.

MicroRNA‑29a (miR‑29a) expression has been reported to be closely associated with skeletal muscle insulin resistance and type 2 . The present study investigated the effect of miR‑29a on (PA)‑induced lipid metabolism dysfunction and insulin resistance in C2C12 myotubes via overexpressing or silencing of miR‑29a expression. Mouse C2C12 myoblasts were cultured, differentiated and transfected with miR‑29a or miR‑29a inhibitor lentiviral with or without subsequent (PA) treatment. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were performed to assess the mRNA and protein levels of related genes, respectively. PA treatment increased the expression of miR‑29a in a time‑ and dose‑ dependent manner. miR‑29a silencing improved insulin‑induced glucose uptake and increased glucose transporter‑4 (GLUT4) transportation to the plasma membrane by upregulating its target peroxisome proliferator‑activated receptor δ (PPARδ). Furthermore, it was observed that miR‑29a regulated the expression of genes associated with lipid metabolism, including pyruvate dehydrogenase kinase isoform, mitochondrial uncoupling protein (UCP)2, UCP3, long chain specific acyl‑CoA dehydrogenase, mitochondrial and fatty transport protein 2. The results confirmed that silencing miR‑29a induced a decrease in glucose transport and affected lipid metabolism in PA‑treated C2C12 cells, and therefore may be involved in insulin resistance by targeting PPARδ in skeletal muscle. Therefore, the inhibition of miR‑29a may be a potential novel strategy for treating insulin resistance and type 2 .

Keyword: diabetes

HAMSCs/HBMSCs coculture system ameliorates osteogenesis and angiogenesis against glucolipotoxicity.

Osteoporosis and vascular lesions induced by glucolipotoxicity are common complications of (DM). In order to deal with these complications, we designed a new therapeutic strategy, i.e. coculture system containing human amnion-derived mesenchymal stem cells (HAMSCs) and human bone marrow mesenchymal stem cells (HBMSCs). Two in\xa0vitro coculture models, transwell and mixed cocultures, were proposed for 7 days with variable HAMSCs: HBMSCs ratios. Then, supernatant from each coculture was used to reverse the deficiency of HBMSCs and human umbilical vein endothelial cells (HUVECs) impaired by high glucose and (GP). We found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation, osteogenic differentiation and superoxide dismutase (SOD) activity, as well as induced the reactive oxygen species (ROS) level in HBMSCs. Meanwhile, glucolipotoxicity suppressed cell proliferation, tube formation capacity and angiogenic potential of HUVECs. Though, HAMSCs/HBMSCs coculture system reduced HBMSCs dysfunction by antioxidant properties and promoted angiogenesis in HUVECs. The mixed HAMSCs/HBMSCs coculture at the optimal ratio of 3/1 showed significantly greater cell proliferation, antioxidant properties, osteogenic and angiogenic differentiation than HBMSCs or HUVECs alone. In conclusion, the current coculture system of HAMSCs/HBMSCs can be a potential therapeutic material for advancing bone and vascular regeneration against DM-induced glucolipotoxicity.Copyright © 2018 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: diabetes

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I , insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: diabetes

BRICHOS domain of Bri2 inhibits islet amyloid polypeptide (IAPP) fibril formation and toxicity in human beta cells.

Aggregation of islet amyloid polypeptide (IAPP) into amyloid fibrils in islets of Langerhans is associated with type 2 , and formation of toxic IAPP species is believed to contribute to the loss of insulin-producing beta cells. The BRICHOS domain of integral membrane protein 2B (Bri2), a transmembrane protein expressed in several peripheral tissues and in the brain, has recently been shown to prevent fibril formation and toxicity of Aβ42, an amyloid-forming peptide in Alzheimer disease. In this study, we demonstrate expression of Bri2 in human islets and in the human beta-cell line EndoC-βH1. Bri2 colocalizes with IAPP intracellularly and is present in amyloid deposits in patients with type 2 . The BRICHOS domain of Bri2 effectively inhibits fibril formation in vitro and instead redirects IAPP into formation of amorphous aggregates. Reduction of endogenous Bri2 in EndoC-βH1 cells with siRNA increases sensitivity to metabolic stress leading to cell death while a concomitant overexpression of Bri2 BRICHOS is protective. Also, coexpression of IAPP and Bri2 BRICHOS in lateral ventral neurons of results in an increased cell survival. IAPP is considered to be the most amyloidogenic peptide known, and described findings identify Bri2, or in particular its BRICHOS domain, as an important potential endogenous inhibitor of IAPP aggregation and toxicity, with the potential to be a possible target for the treatment of type 2 .Copyright © 2018 the Author(s). Published by PNAS.

Keyword: diabetes

Human amnion-derived mesenchymal stem cells promote osteogenesis of human bone marrow mesenchymal stem cells against glucolipotoxicity.

Epidemiological evidence suggests that (DM) is an important factor in promoting periodontitis. It not only affects the attachment of connective tissue but also causes loss of alveolar bone. Hence, there is an urgent need to find an effective treatment for DM-induced bone deficiency. This study aimed to investigate the effects of human amniotic mesenchymal stem cells (HAMSCs) on the proliferation and osteogenic differentiation of DM-induced human bone marrow mesenchymal stem cells (HBMSCs). High glucose and (GP) were used to mimic DM-induced glucolipotoxicity. The proliferation levels were measured using flow cytometry. Alkaline phosphatase activity substrate assays, Alizarin red S staining, and western blotting were used to investigate osteogenic differentiation. Oxidative stress was measured by assaying the levels of reactive oxygen species. This study found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation and osteogenesis, and upregulated the oxidative stress level in HBMSCs. However, HAMSCs attenuated HBMSC dysfunction through antioxidant activity by influencing p38 mitogen-activated protein kinase and vascular endothelial growth factor secretion. In conclusion, our findings indicate that HAMSCs might be suitable for treating DM-mediated bone deficiency.

Keyword: diabetes

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high fat diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related metabolic diseases such as type II (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: diabetes

Activator protein-1 and caspase 8 mediate p38α MAPK-dependent cardiomyocyte apoptosis induced by .

Lipoapoptosis of cardiomyocytes may underlie diabetic cardiomyopathy. Numerous forms of cardiomyopathies share a common end-pathway in which apoptotic loss of cardiomyocytes is mediated by p38α mitogen activated protein kinase (MAPK). Although we have previously shown that (PA), a saturated fatty (SFA) elevated in plasma of type 2 and morbid obesity, induces apoptosis in cardiomyocytes via p38α MAPK-dependent signaling, the downstream cascade events that cause cell death remain unknown. The objective of this study\xa0was to investigate mechanisms involved in -induced cardiomyocyte apoptosis. Human adult ventricular cardiomyocyte line (AC16 cells) exposed to high physiological levels of PA for 16\xa0h showed enhanced transcription and phosphorylation of c-fos and c-jun subunits of AP-1 and transcription of caspase 8. When AC16 cells were transfected with small interfering RNA specific against p38α MAPK (si-p38α) for 24 or 48\xa0h, the amplified phosphorylation of c-fos was dose-dependently attenuated, and procaspase 8 was dose-dependently reduced. With translational knockdown of c-fos, PA-induced apoptosis was diminished. Inhibition of caspase 8 for 24\xa0h reduced apoptosis in PA-treated cardiomyocytes. These findings provide evidence for induction of apoptosis in cardiomyocytes exposed to high SFA by a novel pathway requiring activation of c-fos/AP-1 and caspase 8. These results demonstrate how elevated plasma SFA may lead to continual and cumulative loss of cardiomyocytes and potentially contribute to the\xa0development of diabetic cardiomyopathy.

Keyword: diabetes

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/lipid metabolism in type 2 . Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high-fat diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

Keyword: diabetes

Myristic specifically stabilizes diacylglycerol kinase δ protein in C2C12 skeletal muscle cells.

Decreased levels of the δ isozyme of diacylglycerol kinase (DGK) in skeletal muscle attenuate glucose uptake and, consequently, are critical for the pathogenesis of type 2 . We recently found that free myristic (14:0), but not free (16:0), increased the DGKδ protein levels and enhanced glucose uptake in C2C12 myotube cells. However, it has been unclear how myristic regulates the level of DGKδ2 protein. In the present study, we characterized the myristic -dependent increase of DGKδ protein. A cycloheximide chase assay demonstrated that myristic , but not , markedly stabilized DGKδ protein. Moreover, other DGK isozymes, DGKη and ζ, as well as glucose uptake-related proteins, such as protein kinase C (PKC) α, PKCζ, Akt and glycogen synthase kinase 3β, failed to be stabilized by myristic . Furthermore, DGKδ was not stabilized in cultured hepatocellular carcinoma cells, pancreas carcinoma cells or neuroblastoma cells, and only a moderate stabilizing effect was observed in embryonic kidney cells. A proteasome inhibitor and a lysosome inhibitor, MG132 and chloroquine, respectively, partly inhibited DGKδ degradation, suggesting that myristic prevents, at least in part, the degradation of DGKδ by the ubiquitin-proteasome system and the autophagy-lysosome pathway. Overall, these results strongly suggest that myristic attenuates DGKδ protein degradation in skeletal muscle cells and that this attenuation is fatty -, protein- and cell line-specific. These new findings provide novel insights into the molecular mechanisms of the pathogenesis of type 2 .Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: diabetes

METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, insulin resistance, and . Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: diabetes

Fatty acids, epigenetic mechanisms and chronic diseases: a systematic review.

Chronic illnesses like obesity, type 2 (T2D) and cardiovascular diseases, are worldwide major causes of morbidity and mortality. These pathological conditions involve interactions between environmental, genetic, and epigenetic factors. Recent advances in nutriepigenomics are contributing to clarify the role of some nutritional factors, including dietary fatty acids in gene expression regulation. This systematic review assesses currently available information concerning the role of the different fatty acids on epigenetic mechanisms that affect the development of chronic diseases or induce protective effects on metabolic alterations.A targeted search was conducted in the PubMed/Medline databases using the keywords "fatty acids and epigenetic". The data were analyzed according to the PRISMA-P guidelines.Consumption fatty acids like n-3 PUFA: EPA and DHA, and MUFA: oleic and palmitoleic was associated with an improvement of metabolic alterations. On the other hand, fatty acids that have been associated with the presence or development of obesity, T2D, pro-inflammatory profile, atherosclerosis and IR were n-6 PUFA, saturated fatty acids (stearic and ), and trans fatty acids (elaidic), have been also linked with epigenetic changes.Fatty acids can regulate gene expression by modifying epigenetic mechanisms and consequently result in positive or negative impacts on metabolic outcomes.

Keyword: diabetes

Nrf2-Mediated Antioxidant Defense and Peroxiredoxin 6 Are Linked to Biosynthesis of Ester of 9-Hydroxystearic .

Fatty esters of hydroxy fatty acids (FAHFAs) are lipid mediators with promising antidiabetic and anti-inflammatory properties that are formed in white adipose tissue (WAT) via de novo lipogenesis, but their biosynthetic enzymes are unknown. Using a combination of lipidomics in WAT, quantitative trait locus mapping, and correlation analyses in rat BXH/HXB recombinant inbred strains, as well as response to oxidative stress in murine models, we elucidated the potential pathway of biosynthesis of several FAHFAs. Comprehensive analysis of WAT samples identified ∼160 regioisomers, documenting the complexity of this lipid class. The linkage analysis highlighted several members of the nuclear factor, erythroid 2 like 2 ()-mediated antioxidant defense system (), lipid-handling proteins (), and the family of flavin containing monooxygenases () as the positional candidate genes. Transgenic expression of and deletion of genes resulted in reduction of ester of 9-hydroxystearic (9-PAHSA) and 11-PAHSA levels, while oxidative stress induced by an inhibitor of glutathione synthesis increased PAHSA levels nonspecifically. Our results indicate that the synthesis of FAHFAs via carbohydrate-responsive element-binding protein-driven de novo lipogenesis depends on the adaptive antioxidant system and suggest that FAHFAs may link activity of this system with insulin sensitivity in peripheral tissues.© 2018 by the American Association.

Keyword: diabetes

Multiplex Surface Plasmon Resonance Imaging-Based Biosensor for Human Pancreatic Islets Hormones Quantification.

arises from secretory defects in vascularized micro-organs known as the islets of Langerhans. Recent studies indicated that furthering our understanding of the paracrine effect of somatostatin on glucose-induced insulin secretion could represent a novel therapeutic avenue for . While many research groups are interested in insulin and glucagon secretion, few are particularly focused on studying the paracrine interaction in islets\' cells, and none on monitoring a secretory fingerprint that contemplates more than two hormones. Surface plasmon resonance imaging can achieve high-throughput and multiplexed biomolecule quantification, making it an ideal candidate for detection of multiple islet\'s secretion products if arrays of hormones can be properly implemented on the sensing surface. In this study, we introduced a multiplex surface plasmon resonance imaging-based biosensor for simultaneous quantification of insulin, glucagon, and somatostatin. Performing this multiplex biosensing of hormones was mainly the result of the design of an antifouling sensing surface comprised by a mixed self-assembly monolayer of CHO-PEG-SH and 16-mercaptohexadecanoic , which allowed it to operate in a complex matrix such as an islet secretome. The limit of detection in multiplex mode was 1 nM for insulin, 4 nM for glucagon, and 246 nM for somatostatin with a total analysis time of 21 min per point, making our approach the first reporting a label-free and multiplex measurement of such a combination of human hormones. This biosensor holds the promise of providing us with a mean for the further understanding of the paracrine effect of somatostatin on glucose-induced insulin secretion and consequently help develop novel therapeutic agents for .

Keyword: diabetes

SAD-A, a downstream mediator of GLP-1 signaling, promotes the phosphorylation of Bad S155 to regulate in\xa0vitro β-cell functions.

The incretin hormone GLP-1 reduces β-cell failure in patients with type 2 . Previous studies demonstrated that GLP-1 activates SAD-A, a member of the AMPK family, to regulate glucose-stimulated secretion (GSIS), but the underlying mechanisms of SAD-A regulation of β-cell functions remain poorly understood. Here, we propose that activation of SAD-A by GLP-1 promotes the phosphorylation of Bad S155, which in turn positively affects GSIS and β-cell survival. Bad therefore appears to be a downstream molecule of a SAD-A pathway that mediates the GLP-1-triggered reduction in β-cell failure. Knockdown of endogenous SAD-A expression significantly exacerbated in\xa0vitro β-cell dysfunction under lipotoxic conditions and promoted lipotoxicity-induced apoptosis, whereas overexpression of SAD-A inhibited β-cell apoptosis. SAD-A silencing increased ER stress and inhibited the autophagic flux, which contributed to β-cell apoptosis. Thus, SAD-A appears to function as a downstream molecule of GLP-1 signaling that results in Bad S155 phosphorylation. This phosphorylation might therefore be involved in the GLP-1-linked protection against β-cell dysfunction and apoptosis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

Glucagon-like peptide-1 ameliorates cardiac lipotoxicity in diabetic cardiomyopathy via the PPARα pathway.

Lipotoxicity cardiomyopathy is the result of excessive accumulation and oxidation of toxic lipids in the heart. It is a major threat to patients with . Glucagon-like peptide-1 (GLP-1) has aroused considerable interest as a novel therapeutic target for because it stimulates insulin secretion. Here, we investigated the effects and mechanisms of the GLP-1 analog exendin-4 and the dipeptidyl peptidase-4 inhibitor saxagliptin on cardiac lipid metabolism in diabetic mice (DM). The increased myocardial lipid accumulation, oxidative stress, apoptosis, and cardiac remodeling and dysfunction induced in DM by low streptozotocin doses and high-fat diets were significantly reversed by exendin-4 and saxagliptin treatments for 8\xa0weeks. We found that exendin-4 inhibited abnormal activation of the (PPARα)-CD36 pathway by stimulating protein kinase A (PKA) but suppressing the Rho-associated protein kinase (ROCK) pathway in DM hearts, (PA)-treated rat h9c2 cardiomyocytes (CMs), and isolated adult mouse CMs. Cardioprotection in DM mediated by exendin-4 was abolished by combination therapy with the PPARα agonist wy-14643 but mimicked by PPARα gene deficiency. Therefore, the PPARα pathway accounted for the effects of exendin-4. This conclusion was confirmed in cardiac-restricted overexpression of PPARα mediated by adeno-associated virus serotype-9 containing a cardiac troponin T promoter. Our results provide the first direct evidence that GLP-1 protects cardiac function by inhibiting the ROCK/PPARα pathway, thereby ameliorating lipotoxicity in diabetic cardiomyopathy.© 2018 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

Keyword: diabetes

Saturated fatty acids bound to albumin enhance osteopontin expression and cleavage in renal proximal tubular cells.

Osteopontin (OPN) is one of the proinflammatory cytokines upregulated in the kidneys of diabetic animals and patients with nephropathy. An increase in urinary albumin and albumin-bound fatty acids (FA) presents a proinflammatory environment to the proximal tubules in proteinuric kidney diseases including diabetic nephropathy. This study was designed to examine if FA overload could stimulate OPN expression and cleavage in renal tubule epithelial cells. OPN gene and protein expression was examined in the kidney of Zucker diabetic (ZD) rats and cultured proximal tubular cells exposed to either bovine serum albumin (BSA) or BSA conjugated with (PA), the most abundant saturated plasma FA. Real-time PCR analysis confirmed an upregulation of renal cortical OPN gene correlated with albuminuria and nephropathy progression in ZD rats at the age of 7-20 weeks. Immunofluorescence staining of kidney sections revealed a massive induction of OPN protein in albumin-overloaded proximal tubules of ZD rats. A significant increase in both intact and cleaved OPN proteins was further demonstrated in the diabetic kidney and urine samples, which was attenuated by antiproteinuric treatment with losartan, an angiotensin II receptor blocker. When exposed to fatty -free BSA, NRK-52E cells exhibited an increase in protein levels of full-length and cleaved OPN. Moreover, the increase in OPN fragments was greatly enhanced in the presence of PA (250-500 µM). Together, our results support a stimulatory effect of albumin and conjugated FA on OPN expression and cleavage in renal tubule epithelial cells. Thus, besides lowering albuminuria/proteinuria, mitigating circulating FAs may be an effective intervention for preventing and slowing down the progression of nephropathy associated with obesity and type 2 .

Keyword: diabetes

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

Inflammation markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free fatty acids (FFAs) are known to cause damage in various tissues by inducing inflammation. In this study, we investigated whether a FFA (palmitate) induces inflammation in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive oxygen species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish liver and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including .Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Resveratrol reduces intracellular reactive oxygen species levels by inducing autophagy through the AMPK-mTOR pathway.

Oxidative stress induced by free fatty aggravates endothelial injury, which leads to diabetic cardiovascular complications. Reduction of intracellular oxidative stress may attenuate these pathogenic processes. The dietary polyphenol resveratrol reportedly exerts potential protective effects against endothelial injury. This study determined whether resveratrol can reduce the (PA)-induced generation of reactive oxygen species (ROS) and further explored the underlying molecular mechanisms. We found that resveratrol significantly reduced the PA-induced endothelial ROS levels in human aortic endothelial cells. Resveratrol also induced endothelial cell autophagy, which mediated the effect of resveratrol on ROS reduction. Resveratrol stimulated autophagy via the AMP-activated protein kinase (AMPK)-mTOR pathway. Taken together, these data suggest that resveratrol prevents PA-induced intracellular ROS by autophagy regulation via the AMPK-mTOR pathway. Thus, the induction of autophagy by resveratrol may provide a novel therapeutic candidate for cardioprotection in metabolic syndrome.

Keyword: diabetes

Celastrol Reverses -Induced Insulin Resistance in HepG2 Cells via Restoring the miR-223 and GLUT4 Pathway.

The natural triterpenoid compound celastrol ameliorates insulin resistance (IR) in animal models, but the underlying molecular mechanism is unclear. In this study, we investigated how celastrol regulates IR.The HepG2 cellular IR model was initially established with (PA). The expression and activity of glucose transporter 4 (GLUT4), insulin receptor substrate-1 (IRS1) and 9 microRNAs (miRNAs) (miR-7, -34a, -96, -113, -126, -145, -150, -223 and -370) were detected before and after celastrol treatment using the PA-induced HepG2 IR model.The results showed that 250\u2009µM PA for ≥2 days was optimal for inducing IR in HepG2 cells; 600\u2009nM celastrol significantly attenuated the PA-induced IR in HepG2 cells. The PA-induced GLUT4 and IRS1 downregulation and Ser307 phosphorylation on IRS1 was reversed by subsequent treatment with 600\u2009nM celastrol for 6\u2009h. We next investigated which IR-related miRNAs were possible upstream regulators of celastrol-mediated reversal of PA-induced HepG2 IR. Two miRNAs, miR-150 and -223, were significantly downregulated by PA and were re-raised by subsequent celastrol treatment; and miR-223 was upstream of miR-150. Moreover, knocking down miR-223 abolished celastrol\'s anti-IR effects in the PA-induced model.Collectively, our results demonstrated that celastrol reverses PA-induced IR-related alterations, in part via miR-223 in HepG2 cells. Further investigation is warranted for establishing the clinical potential of celastrol in treating IR-related disorders.Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Dietary modulation of energy homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy metabolism is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: diabetes

TAp63 is correlated with chronic inflammation in patients with newly diagnosed type 2 .

To investigate TAp63 expression in patients with type 2 (T2DM) and the potential correlations between TAp63 and proinflammatory cytokines production and other clinical parameters.Peripheral blood mononuclear cells (PBMCs) and plasma were collected from 72 T2DM (cases) and 72 healthy subjects (controls). Fasting blood glucose (FBG), fasting insulin (FIN) and a blood lipid profile were measured. The homeostasis model assessment (HOMA) was used to estimate insulin resistance (IR). Plasma tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were determined. PBMCs isolated from healthy subjects were cultured with or without 33.3 mmol/l glucose or 0.5 mmol/l (PA) for 6 h, 24 h, 48 h, and 72 h. The expression of TAp63 at mRNA and protein levels in PBMCs was analyzed using real-time qRT-PCR and western blots, respectively.TAp63 expression was significantly lower in T2DM patients compared with that of the controls. In addition, TAp63 expression showed a negative correlation with FBG, FIN, HbA1c, HOMA-IR, FFAs, TNF-α, and IL-6 levels. Treatment with 33.3 mmol/l glucose or 0.5 mmol/l PA increased TAp63 expression in the cultured PBMCs.TAp63 level may be correlated with chronic inflammatory state and perturbed glucose and lipid metabolism in T2DM.Copyright © 2018. Published by Elsevier Inc.

Keyword: diabetes

Hydroxystearic Acids Activate GPR40, Which Is Involved in Their Beneficial Effects on Glucose Homeostasis.

hydroxystearic acids (PAHSAs) are endogenous lipids with anti-diabetic and anti-inflammatory effects. PAHSA levels are reduced in serum and adipose tissue of insulin-resistant people and high-fat diet (HFD)-fed mice. Here, we investigated whether chronic PAHSA treatment enhances insulin sensitivity and which receptors mediate PAHSA effects. Chronic PAHSA administration in chow- and HFD-fed mice raises serum and tissue PAHSA levels ∼1.4- to 3-fold. This improves insulin sensitivity and glucose tolerance without altering body weight. PAHSA administration in chow-fed, but not HFD-fed, mice augments insulin and glucagon-like peptide (GLP-1) secretion. PAHSAs are selective agonists for GPR40, increasing Ca flux, but not intracellular cyclic AMP. Blocking GPR40 reverses improvements in glucose tolerance and insulin sensitivity in PAHSA-treated chow- and HFD-fed mice and directly inhibits PAHSA augmentation of glucose-stimulated insulin secretion in human islets. In contrast, GLP-1 receptor blockade in PAHSA-treated chow-fed mice reduces PAHSA effects on glucose tolerance, but not on insulin sensitivity. Thus, PAHSAs activate GPR40, which is involved in their beneficial metabolic effects.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Resveratrol affects hepatic gluconeogenesis via histone deacetylase 4.

The aim of this study was to determine whether resveratrol (Rev) affects the expression, phosphorylation, and nuclear and cytoplasmic distribution of histone deacetylase 4 (HDAC4), which in turn affects gluconeogenesis in hepatocytes under an insulin-resistant state.HepG2 cells were treated with 0.25 mmol/L (PA) to establish an insulin resistance model. The cells were divided into five groups: control, PA, PA + Rev 100 µM, PA + Rev 50 µM, and PA + Rev 20 µM. After treatment for 24 hours, mRNA and protein expression levels of gluconeogenesis pathway-related molecules and HDAC4 were examined. Next, HepG2 cells were transfected with siRNA-HDAC4. The cells were divided into control, PA, PA + Rev 20 µM, PA + Rev 20 µM +siRNA-HDAC4 negative control, and PA + Rev 20 µM +siRNA-HDAC4 knockdown groups to determine the expression of gluconeogenesis pathway proteins.Compared with the control group, the gluconeogenesis pathway-related molecules, glucose-6-phosphatase catalytic subunit (G6PC), phosphoenolpyruvate carboxykinase 1 (PCK1) and forkhead box protein O1 (FOXO1), were increased, and the phosphorylation of FOXO1 decreased after PA treatment. The p-HDAC4 level decreased with the increase in HDAC4 in the nucleus and the decrease in HDAC4 in the cytoplasm in the PA group. Treatment with Rev 20 µM suppressed gluconeogenesis and promoted HDAC4 shuttling into the cytoplasm from the nucleus. However, 100 and Rev 50 µM exerted the opposite effects. Finally, after HDAC4 knockdown, the expression levels of the key gluconeogenesis molecules, G6PC, PCK1, and FOXO1, were increased, and p-FOXO1 was decreased, indicating that gluconeogenesis was enhanced.A low concentration of Rev inhibited gluconeogenesis under insulin-resistance conditions via translocation of HDAC4 from the nucleus to the cytoplasm.

Keyword: diabetes

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic insulin resistance and Type 2 (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of insulin resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo lipogenesis and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve adipogenesis through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of lipid species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with insulin resistance and risk of developing T2D.

Keyword: diabetes

Mitochondrial dysfunction and inhibition of myoblast differentiation in mice with high-fat-diet-induced pre-.

Pre- is characterized by impaired glucose tolerance (IGT) and/or impaired fasting glucose. Impairment of skeletal muscle function is closely associated with the progression of . However, the entire pathological characteristics and mechanisms of pre- in skeletal muscle remain fully unknown. Here, we established a mouse model of pre-, in which 6-week-old male C57BL6/J mice were fed either normal diet or high-fat diet (HFD) for 8 or 16 weeks. Both non-fasting and fasting glucose levels and the results of glucose and insulin tolerance tests showed that mice fed an 8-week HFD developed pre- with IGT; whereas mice fed a 16-week HFD presented with impaired fasting glucose and impaired glucose tolerance (IFG-IGT). Mice at both stages of pre- displayed decreased numbers of mitochondria in skeletal muscle. Moreover, IFG-IGT mice exhibited decreased mitochondrial membrane potential and ATP production in skeletal muscle and muscle degeneration characterized by a shift in muscle fibers from predominantly oxidative type I to glycolytic type II. Western blotting and histological analysis confirmed that myoblast differentiation was only inhibited in IFG-IGT mice. For primary skeletal muscle satellite cells, inhibition of differentiation was observed in -induced insulin resistance model. Moreover, enhanced myoblast differentiation increased glucose uptake and insulin sensitivity. These findings indicate that pre- result in mitochondrial dysfunction and inhibition of myoblast differentiation in skeletal muscle. Therefore, interventions that enhance myoblast differentiation may improve insulin resistance of at the earlier stage.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Clinopodium chinense Attenuates -Induced Vascular Endothelial Inflammation and Insulin Resistance through TLR4-Mediated NF- B and MAPK Pathways.

Elevated (PA) levels are associated with the development of inflammation, insulin resistance (IR) and endothelial dysfunction. Clinopodium chinense (Benth.) O. Kuntze has been shown to lower blood glucose and attenuate high glucose-induced vascular endothelial cells injury. In the present study we investigated the effects of ethyl acetate extract of C. chinense (CCE) on PA-induced inflammation and IR in the vascular endothelium and its molecular mechanism. We found that CCE significantly inhibited PA-induced toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs). Consequently, this led to the inhibition of the following downstream adapted proteins myeloid differentiation primary response gene 88, Toll/interleukin-1 receptor domain-containing adaptor-inducing interferon- and TNF receptor-associated factor 6. Moreover, CCE inhibited the phosphorylation of Ikappa B kinase , nuclear factor kappa-B (NF- B), c-Jun N-terminal kinase, extracellular regulated protein kinases, p38-mitogen-activated protein kinase (MAPK) and subsequently suppressed the release of tumor necrosis factor- , interleukin-1 (IL-1 ) and IL-6. CCE also inhibited IRS-1 serine phosphorylation and ameliorated insulin-mediated tyrosine phosphorylation of IRS-1. Moreover, CCE restored serine/threonine kinase and endothelial nitric oxide synthase (eNOS) activation and thus increased insulin-mediated nitric oxide (NO) production in PA-treated HUVECs. This led to reverse insulin mediated endothelium-dependent relaxation, eNOS phosphorylation and NO production in PA-treated rat thoracic aortas. These results suggest that CCE can significantly inhibit the inflammatory response and alleviate impaired insulin signaling in the vascular endothelium by suppressing TLR4-mediated NF- B and MAPK pathways. Therefore, CCE can be considered as a potential therapeutic candidate for endothelial dysfunction associated with IR and .

Keyword: diabetes

The Dynamic Effects of Isosteviol on Insulin Secretion and Its Inability to Counteract the Impaired β-Cell Function during Gluco-, Lipo-, and Aminoacidotoxicity: Studies In Vitro.

Isosteviol (ISV), a diterpene molecule, is an isomer of the backbone structure of a group of substances with proven antidiabetic capabilities. The aim of this study was to investigate if ISV elicits dynamic insulin release from pancreatic islets and concomitantly is able to ameliorate gluco-, lipo-, and aminoacidotoxicity in clonal β-cell line (INS-1E) in relation to cell viability and insulin secretion. Isolated mice islets placed into perifusion chambers were perifused with 3.3 mM and 16.7 mM glucose with/without 10 M ISV. INS-1E cells were incubated for 72 h with either 30 mM glucose, 1 mM palmitate or 10 mM leucine with or without 10 M ISV. Cell viability was evaluated with a Cytotoxic Fluoro-test and insulin secretion was measured in Krebs-Ringer Buffer at 3.3 mM and 16.7 mM glucose. In the presence of 3.3 mM glucose, 10 M ISV did not change basal insulin secretion from perifused islets. However, at a high glucose level of 16.7 mM, 10 M ISV elicited a 2.5-fold increase (-ISV: 109.92 ± 18.64 ng/mL vs. +ISV: 280.15 ± 34.97 ng/mL; < 0.01). After 72 h gluco-, lipo-, or aminoacidotoxicity in INS-1E cells, ISV treatment did not significantly affect cell viability (glucotoxicity, -ISV: 19.23 ± 0.83%, +ISV: 18.41 ± 0.90%; lipotoxicity, -ISV: 70.46 ± 3.15%, +ISV: 65.38 ± 2.81%; aminoacidotoxicity: -ISV: 8.12 ± 0.63%; +ISV: 7.75 ± 0.38%, all nonsignificant). ISV did not improve impaired insulin secretion (glucotoxicity, -ISV: 52.22 ± 2.90 ng/mL, +ISV: 47.24 ± 3.61 ng/mL; lipotoxicity, -ISV: 19.94 ± 4.10 ng/mL, +ISV: 22.12 ± 3.94 ng/mL; aminoacidotoxicity: -ISV: 32.13 ± 1.00 ng/mL; +ISV: 30.61 ± 1.54 ng/mL, all nonsignificant). In conclusion, ISV acutely stimulates insulin secretion at high but not at low glucose concentrations. However, ISV did not counteract cell viability or cell dysfunction during gluco-, lipo-, or aminoacidotoxicity in INS-1E cells.

Keyword: diabetes

Primary defects in lipid handling and resistance to exercise in myotubes from obese donors with and without type 2 .

Several studies have shown that human primary myotubes retain the metabolic characteristic of their donors in vitro. Others and we have demonstrated a reduced lipid turnover and fat oxidation rate in myotubes derived from obese donors without and with type 2 (T2D). Because exercise is known to increase fat oxidative capacity in skeletal muscle, we investigated if in vitro exercise could restore primary defects in lipid handling in myotubes of obese without and with T2D compared to lean non-diabetic donors. Primary myotubes cultures were derived from biopsies of lean, obese and T2D subjects. One single bout of long-duration exercise was mimicked in vitro by electrical pulse stimulation (EPS) for 24 h. Lipid handling was measured using radiolabeled palmitate, Metabolic gene expression by real-time qPCR, and proteins by western blot. We first show that myotubes from obese and T2D donors had increased uptake and incomplete oxidation of palmitate. This was associated with reduced mitochondrial respiratory chain complex II, III and IV protein expression in myotubes from obese and T2D subjects. EPS stimulated palmitate oxidation in lean donors, while myotubes from obese and T2D donors were refractory to this effect. Interestingly, EPS increased total palmitate uptake in myotubes from lean donors while myotubes from T2D donors had a reduced rate of palmitate uptake into complex lipids and triacylglycerols. Novelty: • Myotubes from obese and T2D donors are characterized by primary defects in handling. • Both obese and T2D myotubes are partially refractory to the beneficial effect of exercise on lipid handling.

Keyword: diabetes

Antagonistic interaction between Nodal and insulin modulates pancreatic β-cell proliferation and survival.

Insulin signaling pathway in β-cell is essential to promote β-cells proliferation and survival, while Nodal-ALK7-Smad3 signaling involves β-cells apoptosis. We attempted to address inter-relationship between Nodal and insulin in modulating β-cell proliferation and apoptosis.Using INS-1 β-cells and isolated rat islets, we examined the effects of Nodal, insulin, or the two combined on β-cell proliferation and/or apoptosis.The β-cells under high-glucose or palmitate conditions showed significant up-regulation of Nodal expression and activation of its downstream signaling pathway resulted in increased cleaved caspase-3. Insulin treatment led to significantly attenuated Nodal-induced cell apoptotic pathway. Similar results were found in directly Nodal-treated β-cell that insulin could partially block Nodal-induced up-regulation of ALK7-Smad3-caspase-3 signaling pathways with significantly attenuated β-cell apoptosis. Interestingly, we found that insulin-induced Akt activation and downstream molecules including GSK-3β, β-catenin and ERK1/2 was significantly attenuated by the co-treatment with Nodal, resulted in decreased cell proliferation. Furthermore, Nodal decreased glucose-evoked calcium influx and played a negative role during glucose-stimulated insulin secretion in the β-cells. Immunocytochemistry studies showed that Nodal treatment translocated Smad3 from cytosol mostly to the nucleus; however, co-treatment with insulin significantly decreased Smad3 nuclear localization. Co-immunoprecipitation experiments showed a directly interaction between Smad3 and Akt, and this interaction was enhanced by co-treatment with insulin.Our data suggest that the antagonistic interaction between Nodal and insulin has a role in the regulation of β-cell mass and secretion.

Keyword: diabetes

Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.

Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves liver injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 (T2DM) and to determine the underlying mechanisms of these effects.A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with . Major signalling profiles involved in fatty oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry.Nuciferine restored impaired glucose tolerance and insulin resistance in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed -induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation.Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.© 2018 The British Pharmacological Society.

Keyword: diabetes

Low molecular weight fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic fatty-liver disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common liver disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in -induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the liver of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for obesity/-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti-obesity remedy. Several pharmacological studies have shown its beneficial effects in metabolism. Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate lipid and glucose metabolism and are therapeutic targets for type 2 . This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, lipid accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased lipid accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 , correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic insulin resistance development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Sphingomyelinase Down-regulation Alleviates Vascular Endothelial Insulin Resistance in Diabetic Rats.

Insulin resistance in endothelial cells contributes to the development of cardiovascular disease in patients with type 2 . sphingomyelinase (ASM) is a soluble glycoprotein which plays a vital role in the development and progression of various diseases such as cardiovascular and metabolic diseases. However, it remains unknown if ASM regulates insulin resistance in vascular endothelial cells in type 2 . ASM down-regulation with gene silencing and selective inhibitor amitriptyline was used in the rat aortic endothelial cells (RAECs) treated with (PA), a common saturated free fatty , which is thought to be the major cause of insulin resistance. It was shown that ASM down-regulation increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of pIRS-1-ser307 and AKT-ser473 via ceramide, consequently resulting in the decrease of the production of endothelial nitric oxide synthase (eNOS) and nitric oxide in PA-induced RAECs. We further found that ASM down-regulation blocked the Nox2- and Nox4-dependent superoxide (O ) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vivo, amitriptyline relieved the vasodilatory response to acetylcholine and restored the level of ceramide, Nox2 and Nox4 in the aorta endothelium of high-fat diet-fed rats following an injection of streptozotocin. Taken together, these results suggest that ASM down-regulation can improve endothelial insulin resistance which is attributed to inhibiting redox signalling in RAECs. Thus, these data support the idea that ASM is a promising clinical biomarker and potential therapeutic target for diabetic vascular complication.© 2018 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Keyword: diabetes

Saturated fatty combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic inflammation, a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: diabetes

Liraglutide ameliorates palmitate-induced insulin resistance through inhibiting the IRS-1 serine phosphorylation in mouse skeletal muscle cells.

A reduction in insulin-stimulated glucose uptake in skeletal muscles is a characteristic of insulin resistance and type 2 (T2DM). The glucagon-like peptide (GLP)-1 agonist liraglutide can reduce blood glucose levels in individuals with T2DM. However, its effect on insulin-induced glucose metabolism in the skeletal muscle of insulin resistance is unknown. We investigated the effects and action mechanisms of liraglutide on insulin resistance (IR) in the\xa0skeletal muscle cells treatment with (PA).The cell-surface GLUT4myc levels were determined by an antibody-coupled colorimetric assay. The phosphorylation levels of Akt, PI3K(p85α), AS160, IRS1, IKK, and JNK were determined by western blotting. The quantifications of mRNA levels of TNFα, IL-1β, and IL-6 were determined by real-time PCR. Analysis of variance was used for data analysis.PA elevated not only phosphorylation of JNK, IRS1 serines, and IKKα/β, but also the expression of IL-6,\xa0TNFα and IL-1β in C2C12-GLUT4myc cells. PA can reduce phosphorylation of IRS1 tyrosine. These effects of PA were reversed by liraglutide. In addition, liraglutide can reverse PA-decreased insulin-stimulated cell-surface GLUT4 levels, Akt, PI3K(p85α), and AS160 phosphorylation.Liraglutide can enhance insulin-induced GLUT4 translocation by inhibiting IRS1 serine phosphorylation in PA-treated muscle cells.

Keyword: diabetes

Palmitate-Induced Insulin Hypersecretion and Later Secretory Decline Associated with Changes in Protein Expression Patterns in Human Pancreatic Islets.

In obese children with high circulating concentrations of free fatty palmitate, we have observed that insulin levels at fasting and in response to a glucose challenge were several times higher than in obese children with low concentrations of the fatty as well as in lean controls. Declining and even insufficient insulin levels were observed in obese adolescents with high levels of the fatty . In isolated human islets exposed to palmitate we have observed insulin hypersecretion after 2 days exposure. In contrast, insulin secretion from the islets was reduced after 7 days culture in the presence of the fatty . This study aims\xa0at identifying islet-related biological events potentially linked with the observed insulin hypersecretion and later secretory decline in these obese children and adolescents using the islet model. We analyzed protein expression data obtained from human islets exposed to elevated palmitate levels for 2 and 7 days by an improved methodology for statistical analysis of differentially expressed proteins. Protein profiling of islet samples by liquid chromatography-tandem mass spectrometry identified 115 differentially expressed proteins (DEPs). Several DEPs including sorcin were associated with increased glucose-stimulated insulin secretion\xa0in islets after 2 days of exposure to palmitate. Similarly, several metabolic pathways including altered protein degradation, increased autophagy, altered redox condition, and hampered insulin processing were coupled to the functional impairment of islets after 7 days of culture in the presence of palmitate. Such biological events, once validated in the islets, may give rise to novel treatment strategies aiming at normalizing insulin levels in obese children with high palmitate levels, which may reduce or even prevent obesity-related type 2 .

Keyword: diabetes

Hsp74/14-3-3σ Complex Mediates Centrosome Amplification by High Glucose, Insulin, and .

It has been reported recently that type 2 promotes centrosome amplification via 14-3-3σ/ROCK1 complex. In the present study, 14-3-3σ interacting proteins are characterized and their roles in the centrosome amplification by high glucose, insulin, and are investigated. Co-immunoprecipitation in combination with MS analysis identified 134 proteins that interact with 14-3-3σ, which include heat shock 70 kDa protein 4 (Hsp74). Gene ontology analyses reveal that many of them are enriched in binding activity. Kyoto Encyclopedia of Genes and Genomes analysis shows that the top three enriched pathways are ribosome, carbon metabolism, and biosynthesis of amino acids. Molecular and functional investigations show that the high glucose, insulin, and increase the expression and binding of 14-3-3σ and Hsp74 as well as centrosome amplification, all of which are inhibited by knockdown of 14-3-3σ or Hsp74. Moreover, molecular docking analysis shows that the interaction between the 14-3-3σ and the Hsp74 is mainly through hydrophobic contacts and a lesser degree ionic interactions and hydrogen bond by different amino acids residues. In conclusion, the results suggest that the experimental treatment triggers centrosome amplification via upregulations of expression and binding of 14-3-3σ and Hsp74.© 2019 The Authors. Proteomics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: diabetes

Overexpression of heart-type fatty binding protein enhances fatty -induced podocyte injury.

Deregulated lipid metabolism is a characteristic of metabolic diseases including type 2 and obesity, and likely contributes to podocyte injury and end-stage kidney disease. Heart-type fatty binding protein (H-FABP) was reported to be associated with lipid metabolism. The present study investigated whether H-FABP contributes to podocyte homeostasis. Podocytes were transfected by lentiviral vector to construct a cell line which stably overexpressed H-FABP. Small interfering RNA capable of effectively silencing H-FABP was introduced into podocytes to construct a cell line with H-FABP knockdown. Certain groups were treated with (PA) and the fat metabolism, as well as inflammatory and oxidative stress markers were measured. PA accelerated lipid metabolism derangement, inflammatory reaction and oxidative stress in podocytes. Overexpression of H-FABP enhanced the PA-induced disequilibrium in podocytes. The mRNA and protein expression levels of acyl-coenzyme A oxidase 3 and monocyte chemotactic protein 1, and the protein expression levels of 8-hydroxy-2\'-deoxyguanosine and 4-hydroxynonenal were upregulated in the H-FABP overexpression group, while the mRNA and protein expression of peroxisome proliferator activated receptor α was downregulated. Knockdown of H-FABP inhibited the PA-induced injury and lipid metabolism derangement, as well as the inflammatory reaction and oxidative stress in podocytes. These results indicated that overexpression of H-FABP enhances fatty -induced podocyte injury, while H-FABP inhibition may represent a potential therapeutic strategy for the prevention of lipid metabolism-associated podocyte injury.

Keyword: diabetes

Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

Obesity increases the risk of developing . Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in with obesity. Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II . Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling pathways was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of with obesity that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for with obesity.

Keyword: diabetes

Wnt7a promotes wound healing by regulation of angiogenesis and inflammation: Issues on and obesity.

Diabetic skin heals wounds poorly. Though obesity is the common risk factor of , few studies have investigated its effects on wound healing.This study aimed to evaluate the morphology and possible mechanism of human umbilical vein endothelial cells (HUVEC-C) in response to different levels of glucose and , and explore the role of Wnt7a in wound healing.The functional changes of HUVEC-C and mRNA expression of Wnt signaling were determined by analyzing cell viability, migration, tube formation and rt-PCR in gradients of glucose and . Recombinant Wnt7a protein was injected around wounds made on streptozotocin (STZ) -induced diabetic rats with (HF) or without (DM) high-fat diet. Angiogenesis and inflammatory statement were mainly analyzed by immunohistochemistry, ELISA, cytometry and Western blotting.The expression of Wnt7a significantly decreased in high Glc/PA cultured cells or DM and HF wounded rats. Impaired wound healing was also observed in DM and HF groups. The healing rate significantly accelerated after localized injection with Wnt7a at d10. Moreover, the expression of CD31, eNOS phosphorylation and NO were increased; the reduction of local neutrophils influx, ICAM-1 and IL-6/8 expression levels were obvious especially in diabetic with obesity rats at d10 after Wnt7a treatment.This study indicates the potential role of Wnt7a, which is beneficial for regeneration of damaged vessels, moderation of inflammatory statement in diabetic wound healing with or without obesity, thus demonstrating its possible utility as a topical administration to promote healing rate.Copyright © 2018. Published by Elsevier B.V.

Keyword: diabetes

Consumption of a high fat diet promotes protein O-GlcNAcylation in mouse retina via NR4A1-dependent GFAT2 expression.

The incidence of type 2 , the most common cause of diabetic retinopathy (DR), is rapidly on the rise in developed countries due to overconsumption of calorie rich diets. Using an animal model of diet-induced obesity/pre-, we evaluated the impact of a diet high in saturated fat (HFD) on O-GlcNAcylation of retinal proteins, as dysregulated O-GlcNAcylation contributes to diabetic complications and evidence supports a role in DR. Protein O-GlcNAcylation was increased in the retina of mice fed a HFD as compared to littermates receiving control chow. Similarly, O-GlcNAcylation was elevated in retinal Müller cells in culture exposed to the saturated fatty palmitate or the ceramide analog Cer6. One potential mechanism responsible for elevated O-GlcNAcylation is increased flux through the hexosamine biosynthetic pathway (HBP). Indeed, inhibition of the pathway\'s rate-limiting enzyme glutamine-fructose-6-phosphate amidotransferase (GFAT) prevented Cer6-induced O-GlcNAcylation. Importantly, expression of the mRNA encoding GFAT2, but not GFAT1 was elevated in both the retina of mice fed a HFD and in retinal cells in culture exposed to palmitate or Cer6. Notably, expression of nuclear receptor subfamily 4 group A member 1 (NR4A1) was increased in the retina of mice fed a HFD and NR4A1 expression was sufficient to promote GFAT2 mRNA expression and O-GlcNAcylation in retinal cells in culture. Whereas palmitate or Cer6 addition to culture medium enhanced NR4A1 and GFAT2 expression, chemical inhibition of NR4A1 transactivation repressed Cer6-induced GFAT2 mRNA expression. Overall, the results support a model wherein HFD increases retinal protein O-GlcNAcylation by promoting NR4A1-dependent GFAT2 expression.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Observational clinical and nerve conduction study on effects of a nutraceutical combination on painful diabetic distal symmetric sensory-motor neuropathy in patients with type 1 and type 2.

Painful distal symmetric polyneuropathy (pDSPN) is one of the most common and invalidating complications of , both of type 1 and type 2. Mechanisms responsible for the occurrence of the pDSPN are multifactorial and involve metabolic pathways regulating inflammation, microvessel circulation, axonal degeneration and so on. Several therapeutic approaches have been proposed to treat pain and each of them showed positive effects associated to drug-related side effects.Twenty-five consecutive patients with diagnosis of and pDSPN and tried to manage pain with a dietary supplement composed of a mixture of natural extracts (β-caryophyllene, myrrh, carnosic ) and PEA. This is a nutraceutical with potential multiple effects on metabolic, pain and vascular compartments, a profile considered useful in pDSPN. Patients were enrolled and polyneuropathy evaluated by means of nerve conduction study. Pain was assessed using VAS score scale and MNSI. Each patient was evaluated at T0 (time of enrollment) and at T1 (after 16 weeks of treatment).Supplement administration was well tolerated and induced unexpectedly significant amelioration of polyneuropathy with increase amplitude and reduction of pain. No side effects were reported.This fixed combination could well be considered as a potential nutraceutical option to manage pDSPN in diabetic patients.

Keyword: diabetes

PAQR3 regulates phosphorylation of FoxO1 in insulin-resistant HepG2 cells via NF-κB signaling pathway.

Insulin resistance is a significant feature of type 2 and glucose and lipid metabolism disorders. Activation of NF-κB signaling pathway plays an important role in the formation of insulin resistance. FoxO1 plays a major role in regulating glucose and lipid metabolism, as well as insulin signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under insulin resistance. In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced insulin-resistant HepG2 cells, thereby causing glucose and lipid metabolism disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced insulin-resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and lipid metabolism disorders induced by insulin resistance.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: diabetes

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: diabetes

Nutraceutical Potential of in Metabolic Syndrome.

L. is a well-known fruit worldwide, and its highest production occurs in tropical and subtropical regions. The pulp contains vitamins A, C, and E, B complex vitamins, such as pantothenic and folate, and minerals, such as magnesium and potassium, as well as food fibers. Phenolic compounds, such as benzyl isothiocyanate, glucosinolates, tocopherols (α and δ), β-cryptoxanthin, β-carotene and carotenoids, are found in the seeds. The oil extracted from the seed principally presents oleic fatty followed by , linoleic and stearic acids, whereas the leaves have high contents of food fibers and polyphenolic compounds, flavonoids, saponins, pro-anthocyanins, tocopherol, and benzyl isothiocyanate. Studies demonstrated that the nutrients present in its composition have beneficial effects on the cardiovascular system, protecting it against cardiovascular illnesses and preventing harm caused by free radicals. It has also been reported that it aids in the treatment of and in the reduction of cholesterol levels. Thus, both the pulp and the other parts of the plant (leaves and seeds) present antioxidant, anti-hypertensive, hypoglycemic, and hypolipidemic actions, which, in turn, can contribute to the prevention and treatment of obesity and associated metabolic disorders.

Keyword: diabetes

Erythrocyte Saturated Fatty Acids and Incident Type 2 in Chinese Men and Women: A Prospective Cohort Study.

The association between circulating saturated fatty acids (SFAs) and incident type 2 (T2D) is reported in Western populations with inconsistent results, while evidence from Asian populations is scarce. We aimed to examine the associations between erythrocyte SFAs and incident T2D in a Chinese population. Between 2008 and 2013, a total of 2683 participants, aged 40⁻75 years, free of were included in the present analyses. Incident T2D cases were ascertained during follow-up visits. Gas chromatography was used to measure erythrocyte fatty acids at baseline. The Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs). During 13,508 person years of follow-up, 216 T2D cases were identified. Compared with the first quartile, multivariable-adjusted HRs (95% CIs) of the fourth quartile were 1.20 (0.82⁻1.76; = 0.242) for myristic (14-carbon tail, zero double bonds; 14:0), 0.69 (0.48⁻0.99; = 0.080) for (16:0), 1.49 (1.02⁻2.19; = 0.047) for stearic (18:0), 1.46 (1.00⁻2.12; = 0.035) for arachidic (20:0), 1.48 (0.99⁻2.22; = 0.061) for behenic (22:0), and 1.08 (0.74⁻1.56; = 0.913) for lignoceric (24:0). Our findings indicate that individual erythrocyte SFAs are associated with T2D in different directions, with 18:0 and 20:0 SFAs positively associated with the risk, whereas no convincing inverse association for 16:0 SFAs.

Keyword: diabetes

Phlorizin Exerts Direct Protective Effects on (PA)-Induced Endothelial Dysfunction by Activating the PI3K/AKT/eNOS Signaling Pathway and Increasing the Levels of Nitric Oxide (NO).

BACKGROUND Sodium glucose transporter-2 inhibitors are the newest antidiabetic drugs that seem to be cardioprotective and can prevent type 2 in patients with high cardiovascular risks. Previous clinical trials have shown that these inhibitors can alleviate endothelial dysfunction, but the mechanism of action remains unknown. How SGLT inhibitor influences the release of NO in PA-induced HUVECs has never been reported. MATERIAL AND METHODS To explore the potential effects of the endothelial-protective mechanism of phlorizin and its impact on nitric oxide (NO), human umbilical vein endothelial cells (HUVECs) were incubated with (PA) and then treated with phlorizin. Western blotting was performed to assess the phosphorylation of AKT, eNOS, and IRS-1. To further explore potential targets, siRNA transfection was used to demonstrate the role of SGLT1 and SGLT2. RESULTS Phlorizin suppressed the expression of SGLT1 and SGLT2, activated the PI3K/AKT/eNOS signaling pathway, increased the output of NO, and promoted the consumption of glucose in PA-induced HUVECs. Through demonstrating siRNA suppression of the expression of SGLT1 and SGLT2 in PA-induced HUVECs, this study provides a new understanding of the mechanism behind SGLT1 and SGLT2. CONCLUSIONS Our data demonstrate that phlorizin ameliorates the endothelial dysfunction link with the activation of the PI3K/AKT/eNOS signaling pathway and augmentation of the release of NO, partially through suppressing the expression of SGLT1 and SGLT2 in PA-induced HUVECS.

Keyword: diabetes

Secreted Wnt6 mediates -associated centrosome amplification via its receptor FZD4.

We have recently published that type 2 promotes cell centrosome amplification via up-regulation of ROCK1 and 14-3-3σ. The study further investigated the molecular mechanisms underlying the -associated centrosome amplification. We found that treatment of cells with high glucose, insulin and increased the intracellular and the extracellular protein levels of Wnt6 as well as the cellular level of β-catenin. The treatment also activated β-catenin and promoted its nuclear translocation. Treatment of cells with siRNA species for Wnt6, FZD4 or β-catenin as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture medium could all attenuate the treatment-triggered centrosome amplification. Moreover, we showed that secreted Wnt6-FZD4-β-catenin was the signaling pathway which was upstream of ROCK1 and 14-3-3σ. We also found that AGEs were also able to increase the cellular and extracellular levels of Wnt6, the cellular protein level of β-catenin, and centrosome amplification. Treatment of the cells with siRNA species for Wnt6 or FZD4 as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture could all inhibit the AGEs-elicited centrosome amplification. In colon tissues from diabetic mice model, the protein levels of Wnt6 and 14-3-3σ were increased. In conclusion, our results showed that the pathophysiological factors in type 2 , including AGEs, were able to induce centrosome amplification. It is suggested that secreted Wnt6 binds to FZD4 to activate the canonical Wnt6 signaling pathway which is upstream of ROCK1 and 14-3-3σ, which is the cell signaling pathway underlying the -associated centrosome amplification.

Keyword: diabetes

Risk of associated with fatty acids in the de novo lipogenesis pathway is independent of insulin sensitivity and response: the Insulin Resistance Atherosclerosis Study (IRAS).

To examine the associations of fatty acids in the de novo lipogenesis (DNL) pathway, specifically myristic (14:0), (16:0), palmitoleic (c16:1\u2009n-7), myristoleic (c14:1n5), stearic (18:0) and oleic (c18:1\u2009n-9), with 5-year risk of type 2 . We hypothesized that DNL fatty acids are associated with risk of type 2 independent of insulin sensitivity.We evaluated 719 (mean age 55.1±8.5 years, 44.2% men, 42.3% Caucasians) participants from the Insulin Resistance Atherosclerosis Study. Multivariable logistic regression models with and without adjustment of insulin sensitivity were used to assess prospective associations of DNL fatty acids with incident type 2 .Type 2 incidence was 20.3% over 5\u2009years. In multivariable regression models, , palmitoleic, myristic, myristoleic and oleic acids were associated with increased risk of type 2 (p<0.05). had the strongest association (OR per standard unit of 1.46; 95%\u2009CI 1.23 to 1.76; p<0.001), which remained similar with addition of insulin sensitivity and acute insulin response (AIR) to the model (OR 1.36; 95%\u2009CI 1.09 to 1.70, p=0.01). Oleic and palmitoleic acids were also independently associated with incident type 2 . In multivariable models, ratios of fatty acids corresponding to stearoyl CoA desaturase-1 and Elovl6 enzymatic activity were significantly associated with risk of type 2 independent of insulin sensitivity and AIR.We observed associations of DNL fatty acids with type 2 incidence independent of insulin sensitivity.

Keyword: diabetes

N-3 Polyunsaturated Fatty Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Rats by Postponing Shortening of Hepatic Telomeres and Modulating Liver Metabolism.

The long-term influence of gestational (GDM) on offspring and the effect of omega-3 polyunsaturated fatty acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and inflammation in the liver of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the liver, in which numerous metabolites and metabolic pathways were altered when GDM offspring grew to old age. Many metabolites were related to risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: diabetes

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low fat diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic lipid accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and , and (3) dietary SFA type impacts LCPUFA metabolism.Published by Elsevier Inc.

Keyword: diabetes

Hybrid lipids, peptides, and lymphocytes: new era in type 1 research.

Type 1 (T1D) results from autoimmune destruction of insulin-producing β cells in islets of Langerhans. Many genetic and immunological insights into autoimmune disease pathogenesis were initially uncovered in the context of T1D and facilitated by preclinical studies using the nonobese diabetic (NOD) mouse model. Recently, the study of T1D has led to the discovery of fatty esters of hydroxyl fatty acids (FAHFAs), which are naturally occurring hybrid peptides that modulate inflammation and pathogenesis, and a hybrid lymphocyte that expresses both B and T cell receptors. esters of hydroxy stearic acids (PAHSAs) are the most extensively studied FAHFA. In this issue of the JCI, Syed et al. have shown that PAHSAs both attenuate autoimmune responses and promote β cell survival in NOD mice. Given the lack of effective T1D therapies and the paucity of known side effects of PAHSAs, this lipid may have therapeutic potential for individuals at risk for or newly diagnosed with T1D.

Keyword: diabetes

PPARγ promotes -associated centrosome amplification via increasing the expression of SKA1 directly at the transcriptional level.

We have\xa0recently published that type 2 can induce cell centrosome amplification due to the action of high glucose, , and insulin, and ROCK1 and 14-3-3σ are signal mediators. In this study, we further investigated the molecular mechanisms of the centrosome amplification in colon cancer HCT116 cells. Treatment of the cells with high glucose, , and insulin increased the expression of peroxisome proliferator-activated receptor γ (PPARγ) as well as the spindle and kinetochore associated protein 1 (SKA1), knockdown of each of which resulted in the inhibition of the treatment-triggered centrosome amplification. Knockdown of PPARγ inhibited the treatment-evoked increase in the SKA1 level, whereas knockdown of SKA1 did not modify the treatment-increased PPARγ level. We found a predicted binding site for PPARγ in the promoter region of the SKA1 gene from the JASPAR database. Experimental results showed that the treatment increased the messenger RNA level of SKA1, which could be inhibited by PPARγ chemical inhibitor or small interfering RNA. Moreover, we were able to show that PPARγ could bind to the binding site in the SKA1 gene promoter, which was increased by the experimental treatment. In conclusion, it is suggested that the pathophysiological factors in type 2 , high glucose, , and insulin, induce the cell centrosome amplification through the PPARγ-SKA1 pathway, in which PPARγ increases the expression of SKA1 via directly enhancing the SKA1 gene transcription.© 2019 Wiley Periodicals, Inc.

Keyword: diabetes

Peroxidase expression is decreased by palmitate in cultured podocytes but increased in podocytes of advanced diabetic nephropathy.

High levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 . (PA) is the predominant circulating saturated FFA. PA induces mitochondrial superoxide and hydrogen peroxide (H O ) generation in cultured podocytes. To elucidate the role of PA in antioxidant defense systems in diabetic nephropathy (DN), cultured podocytes were exposed to 250\u2009μM PA for 1-24\u2009hr, and protein expressions of catalase, peroxiredoxins (Prxs), and glutathione peroxidase (GPx) were examined by western blot analysis. PA induced an early transient increase in the Prx1, Prx2, and GPx1 levels in podocytes, but not catalase. Long-term exposure of PA to podocytes significantly decreased the protein levels of Prx1, Prx2, GPx1, and catalase. Coincubation of PA-treated cells with oleic , however, restored the expression of these proteins. In advanced human diabetic glomeruli, H O generation was elevated as shown by increased fluorescence of dichlorofluorescein. Strong immunostaining for Prx1, Prx2, GPx1, and catalase was observed in the podocytes of advanced human DN, wherein transforming growth factor-β1 staining was also positive. These results suggest that podocytes are susceptible to PA-induced oxidative damage with impaired peroxidase activity and that peroxidases have futile antioxidant effects in the podocytes in the late stages of DN. Given this, PA-induced podocyte injury via inadequate peroxidase response to H O appears to play an important role in the pathogenesis of DN.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Lipid profile changes in erythrocyte membranes of women with diagnosed GDM.

Gestational (GDM) is a glucose intolerance that begins or is first recognized during pregnancy. It is currently a growing health problem worldwide affecting from 1% to 14% of all pregnant women depending on racial and ethnic group as well as the diagnostic and screening criteria. Our preliminary study aimed at investigating the erythrocyte membrane fatty profiles of pregnant women, in particular with diagnosed with gestational (GDM), and with normal glucose tolerant (NGT) pregnant women as a control group. The study group comprised 43 pregnant women, 32 of whom were diagnosed with GDM according to the WHO criteria, and 11 with normal glucose tolerance. The erythrocyte membrane phospholipids were obtained according to the Folch extraction procedure. Fatty acids (FA) were analyzed by gas chromatography (GC) as the corresponding fatty methyl esters (FAME). A cluster of 14 fatty acids identified contained >98% of the recognized peaks in the GC analysis. The analysis of fatty acids from erythrocytes revealed important differences between GDM and NGT women in the third trimester, and the results were correlated with biochemical data. Among the 14 measured FA representing the membrane lipidomic profile, the levels of three saturated FA (myristic, , stearic acids) tended to decrease in GDM patients, with the percentage content of stearic significantly changed. The relative content of monounsaturated fatty acids (MUFA) tended to increase, in particular the oleic and vaccenic contents were significantly increased in erythrocyte membranes of the GDM group in comparison with the NGT group. The GDM group demonstrated higher sapienic levels (+29%) but this change was not statistically significant. This study revealed association between an impaired cis-vaccenic concentration in erythrocytes membrane and GDM development. No significant changes of polyunsaturated fatty acids (PUFA) were observed in GDM and NGT erythrocytes. We postulate, basing on the differences between the GDM and NGT lipidomic profiles, that stearic and cis-vaccenic acids can be considered as dual biomarkers of specific SFA-MUFA conversion pathway, involving the coupling of delta-9 desaturase and elongase enzymes. Our results indicate that the SFA-MUFA families may be involved in the pathophysiology of metabolic diseases such as GDM, but the further studies are needed to confirm our hypothesis. In conclusion, the erythrocyte membranes of GDM women undergo remodeling resulting in abnormal fatty profiles, which are reflection of the long-term status of organism and can have great impact on both the mother and her offspring.

Keyword: diabetes

Vernonia amygdalina Delile extract inhibits the hepatic gluconeogenesis through the activation of adenosine-5\'monophosph kinase.

It has been reported that Vernonia amygdalina Delile(VA) presents an anti-diabetic effect, and the effect of VA on lowering glucose is formulated via suppressing the expression of the key hepatic gluconeogenesis enzyme. Therefore, we further explored the probable mechanism of VA on dismissing hepatic gluconeogenesis through the activation of adenosine-5\' monophosphate kinase (AMPK) in vivo and in vitro.We developed type 2 diabetic mice with STZ and oral administration with VA (50\u202fmg/kg, 100\u202fmg/kg and 150\u202fmg/kg) once a day for 6 weeks. Fasting blood glucose (FBG), fasting insulin (FINS) and oral glucose tolerance tests (OGTT) were conducted. The expression levels of AMPK, phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase) proteins in live were evaluated by western blot. Then, we further explored the mechanism of VA on hepatic gluconeogenesis in vitro experiments. Glucose production and the expression of AMPK, PEPCK and G6Pase proteins were detected after VA treatment with the presence of the AMPK inhibitor Compound C.VA reduced FBG and caused a significant improvement in glucose tolerance and insulin resistance (HOMA-IR) in STZ-induced mice. VA inhibited the elevated expression of gluconeogenesis key enzymes (PEPCK and G6Pase) and up-regulated AMPK activity in liver. In (PA)-induced HepG2 cells, VA decreased glucose production and the expression of PEPCK and G6Pase proteins, also activated AMPK pathway. The effects of VA on gluconeogenesis could be reversed by Compound C.These results reveal that VA suppresses hepatic gluconeogenesis at least partially through activating the AMPK.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Patients: A Pilot Study.

Metabolic changes in insulin-dependent (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Association. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: diabetes

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial lipid accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic lipid metabolism. Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 animal models. After streptozotocin (STZ) or high-fat diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, lipid accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and lipid accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing lipid accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Synthesis and biological evaluations of marine oxohexadecenoic acids: PPARα/γ dual agonism and anti-diabetic target gene effects.

Obesity and associated disorders such as metabolic syndrome and type 2 (T2D) have reached epidemic proportions. Several natural products have been reported as Peroxisome Proliferator-Activated Receptor (PPAR) agonists, functioning as lead compounds towards developing new anti-diabetic drugs due to adverse side effects of existing PPAR drugs. We recently isolated and identified (7E)-9-oxohexadec-7-enoic (1) and (10E)-9-oxohexadec-10-enoic (2) from the marine algae Chaetoceros karianus. Herein we report the total synthesis, pharmacological characterization, and biological evaluations of these naturally occurring oxo-fatty acids (oFAs). The syntheses of 1 and 2 afforded sufficient material for extensive biological evaluations. Both oFAs show an appreciable dose-dependent activation of PPARα and -γ, with EC values in the micromolar range, and an ability to regulate important PPAR target genes in hepatocytes and adipocytes. Moreover, both 1 and 2 are able to drive adipogenesis when evaluated in the Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocyte cell model, but with lowered expression of adipocyte markers and reduced lipid accumulation compared to the drug rosiglitazone. This seems to be caused by a transient upregulation of PPARγ and C/EBPα expression. Importantly, whole transcriptome analysis shows that both compounds induce anti-diabetic gene programs in adipocytes by upregulating insulin-sensitizing adipokines and repressing pro-inflammatory cytokines.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: diabetes

Alkannin Inhibited Hepatic Inflammation in Diabetic Db/Db Mice.

The current study was designed to investigate the protective role of alkannin (ALK) on liver injury in diabetic C57BL/KsJ-db/db mice and explore its potential mechanisms.An oral glucose tolerance test (OGTT) was performed. The levels of insulin, alanine aminotransferase (ALT), aspartate aminotransaminase (AST), total cholesterol (TC) and triglyceride (TG) were determined by commercial kits. The pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were determined by ELISA. The levels of the ROCK/NF-κB pathway were determined by Western blotting.The contents of pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were inhibited by ALK, metformin or fasudil in diabetic db/db mice. Further, Western blotting analysis showed that the expression of Rho, ROCK1, ROCK2, p-NF-κBp65, and p-IκBα was significantly reversed by ALK treatment. In human hepatic HepG2 cells, the hepatoprotective effects of ALK were further characterized. With response to -challenge, increased amounts of insulin, ALT, AST, TG, and TC were observed, whereas ALK pretreatment significantly inhibited their leakage in HepG2 cells without appreciable cytotoxic effects. The inflammation condition was recovered with ALK treatment as shown by changes of IL-1β, IL-6 and TNF-α. Further, Western blotting analysis also suggested that ALK improves hepatic inflammation in a Rho-kinase pathway.The present study successfully investigated the role of Rho-kinase signalling in diabetic liver injury. ALK exhibited hepatoprotective effects in diabetic db/db mice, and it might act through improving hepatic inflammation through the Rho-kinase pathway.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Astragalosides IV protected the renal tubular epithelial cells from free fatty acids-induced injury by reducing oxidative stress and apoptosis.

Renal tubular injury is associated with the development of diabetic nephropathy (DN) and the end-stage renal disease (ESRD). Free fatty acids (FFAs)-associated lipotoxicity contributes to injury of proximal renal tubular epithelial (HK-2) cells in . (PA) which is the most abundant saturated fatty in FFAs is closely associated with the gradual decline of renal function. Astragalosides IV (AS-IV) has a variety of pharmacological effects such as anti-inflammation and anti-oxidation. In the current study, we investigated the effects of AS-IV on PA-induced apoptosis of HK-2 cells and the underlying mechanisms. The results showed that AS-IV (10, 20, 40\u2009μmol/L) could alleviate PA-induced apoptosis of HK-2 cells. We found that AS-IV reduced the expression of Bax and cleaved-caspase3, but increased the expression of Bcl-2 and phosphorylated Nrf2 in HK-2 cells. Moreover, AS-IV reduced the level of reactive oxygen species (ROS) in the cells. Our study suggests that AS-IV could protect against PA-induced apoptosis in HK-2 cells by inhibiting ROS generation and apoptotic protein expression. This study may provide a new theoretical option for the patients with type 2 .Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: diabetes

Nucleophosmin/B23 contributes to hepatic insulin resistance through the modulation of NF-κB pathway.

Nucleophosmin (NPM)/B23 is an important nucleolar phosphoprotein involved in the regulation of assorted cellular signaling pathways. In the present study, we revealed a critical role of NPM in liver insulin resistance. NPM is markedly upregulated in insulin-resistant liver tissues and (PA)-exposed HepG2 cells both at mRNA and protein levels. Ectopic expression of NPM in hepatocytes aggravated PA-induced insulin resistance, lipid droplet accumulation, glucose intake impairment as well as the expression of gluconeogenic genes. Coinciding with these results, interference of NPM using small interfering RNA (siRNA) oligos ameliorated PA-induced insulin resistance, as revealed by increased phosphorylation of AKT and GSK3β following insulin treatment. As predicted, PA-triggered alterations in glucose intake and the expression of gluconeogenic enzymes were attenuated following NPM depletion. Finally, we showed that NPM plays an indispensible role in PA-induced activation of NF-κB pathway. Both of NF-κB p65 phosphorylation and nuclear translocation were impeded by NPM interference in PA-treated HepG2 cells. Taken together, these findings explicitly demonstrate that NPM participates in the development of liver insulin resistance, suggesting that NPM may serve as a potential therapeutic target of type 2 .Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Attenuation of Free Fatty -Induced Muscle Insulin Resistance by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 . Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties.

Keyword: diabetes

PAHSAs attenuate immune responses and promote β cell survival in autoimmune diabetic mice.

esters of hydroxy stearic acids (PAHSAs) are endogenous antidiabetic and antiinflammatory lipids. Here, we show that PAHSAs protect against type 1 (T1D) and promote β cell survival and function. Daily oral PAHSA administration to nonobese diabetic (NOD) mice delayed the onset of T1D and markedly reduced the incidence of T1D, whether PAHSAs were started before or after insulitis was established. PAHSAs reduced T and B cell infiltration and CD4+ and CD8+ T cell activation, while increasing Treg activation in pancreata of NOD mice. PAHSAs promoted β cell proliferation in both NOD mice and MIN6 cells and increased the number of β cells in NOD mice. PAHSAs attenuated cytokine-induced apoptotic and necrotic β cell death and increased β cell viability. The mechanism appears to involve a reduction of ER stress and MAPK signaling, since PAHSAs lowered ER stress in NOD mice, suppressed thapsigargin-induced PARP cleavage in human islets, and attenuated ERK1/2 and JNK1/2 activation in MIN6 cells. This appeared to be mediated in part by glucagon-like peptide 1 receptor (GLP-1R) and not the G protein-coupled receptor GPR40. PAHSAs also prevented impairment of glucose-stimulated insulin secretion and improved glucose tolerance in NOD mice. Thus, PAHSAs delayed the onset of T1D and reduced its incidence by attenuating immune responses and exerting direct protective effects on β cell survival and function.

Keyword: diabetes

PCNA and JNK1-Stat3 pathways respectively promotes and inhibits -associated centrosome amplification by targeting at the ROCK1/14-3-3σ complex in human colon cancer HCT116 cells.

We have recently reported that type 2 promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho-associated coiled-coil containing protein kinase 1 (ROCK1)/14-3-3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14-3-3σ. Individual knockdown of PCNA, NPM, or 14-3-3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment-increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and also increased the expressions of c-Jun N-terminal kinase-1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment-increased expression of 14-3-3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14-3-3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment-increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1-Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14-3-3σ complex, and NPM serves as an independent signal for the centrosome amplification.© 2018 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

Keyword: diabetes

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid metabolism that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty metabolism and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty metabolism. Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: diabetes

Cellular toxicity of dietary trans fatty acids and its correlation with ceramide and diglyceride accumulation.

High fatty (FA) levels are deleterious to pancreatic β-cells, largely due to the accumulation of biosynthetic lipid intermediates, such as ceramides and diglycerides, which induce ER stress and apoptosis. Toxicity of palmitate (16:0) and oleate (18:1 cis-Δ) has been widely investigated, while very little data is available on the cell damages caused by elaidate (18:1 trans-Δ) and vaccenate (18:1 trans-Δ), although the potential health effects of these dietary trans fatty acids (TFAs) received great publicity. We compared the effects of these four FAs on cell viability, apoptosis, ER stress, JNK phosphorylation and autophagy as well as on ceramide and diglyceride contents in RINm5F insulinoma cells. Similarly to oleate and unlike palmitate, TFAs reduced cell viability only at higher concentration, and they had mild effects on ER stress, apoptosis and autophagy. Palmitate increased ceramide and diglyceride levels far more than any of the unsaturated fatty acids; however, incorporation of TFAs in ceramides and diglycerides was strikingly more pronounced than that of oleate. This indicates a correlation between the accumulation of lipid intermediates and the severity of cell damage. Our findings reveal important metabolic characteristics of TFAs that might underlie a long term toxicity and hence deserve further investigation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: diabetes

Complement Component C3 Is Highly Expressed in Human Pancreatic Islets and Prevents β Cell Death via ATG16L1 Interaction and Autophagy Regulation.

We show here that human pancreatic islets highly express C3, which is both secreted and present in the cytosol. Within isolated human islets, C3 expression correlates with type 2 (T2D) donor status, HbA1c, and inflammation. Islet C3 expression is also upregulated in several rodent models. C3 interacts with ATG16L1, which is essential for autophagy. Autophagy relieves cellular stresses faced by β cells during T2D and maintains cellular homeostasis. C3 knockout in clonal β cells impaired autophagy and led to increased apoptosis after exposure of cells to and IAPP. In the absence of C3, autophagosomes do not undergo fusion with lysosomes. Thus, C3 may be upregulated in islets during T2D as a cytoprotective factor against β cell dysfunction caused by impaired autophagy. Therefore, we revealed a previously undescribed intracellular function for C3, connecting the complement system directly to autophagy, with a broad\xa0potential importance in other diseases and cell types.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high-fat diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the liver, liver/body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high-fat diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high-fat diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Ceramide stearic to ratio predicts incident .

Ceramide lipids have a role in the development of insulin resistance, and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident .A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the metabolic syndrome (Prevent Metabolic Syndrome [PrevMetSyn], n\u2009=\u2009371). risk score models were developed to estimate the 10\xa0year risk of incident .Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and risk scores significantly decreased in individuals who had 5% or more weight loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident , and may be modulated by lifestyle intervention.

Keyword: diabetes

Acute and Repeated Treatment with 5-PAHSA or 9-PAHSA Isomers Does Not Improve Glucose Control in Mice.

Fatty esters of hydroxylated fatty acids (FAHFAs) were discovered as a novel class of endogenous mammalian lipids whose profound effects on metabolism have been shown. In the current study, in\xa0vitro and in\xa0vivo the metabolic effects of two of these FAHFAs, namely -5- (or -9) -hydroxy-stearic (5- or 9-PAHSA, respectively) were profiled. In DIO mice fed with differentially composed low- or high-fat diets, acute and subchronic treatment with 5-PAHSA and 9-PAHSA alone, or in combination, did not significantly improve the deranged metabolic status. Neither racemic 5- or 9-PAHSA, nor the enantiomers were able to: (1) increase basal or insulin-stimulated glucose uptake in\xa0vitro, (2) stimulate GLP-1 release from GLUTag cells, or (3) induce GSIS in rat, mouse, or human islets or in a human pancreatic β cell line. Therefore, our data do not support the\xa0further development of PAHSAs or their derivatives for the control of insulin resistance and hyperglycemia.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Kaempferol alleviates -induced lipid stores, endoplasmic reticulum stress and pancreatic β-cell dysfunction through AMPK/mTOR-mediated lipophagy.

Kaempferol, a natural flavonoid, has the beneficial effects of preserving pancreatic β-cell mass and function, but its action on β-cell lipid metabolism still remains elusive. Recently, autophagy has been reported to play a major role in lipid metabolism in various cell types, but its role in pancreatic β-cell\'s lipid metabolism is rarely reported. Here, we investigated the role of kaempferol-induced autophagy in inhibition of lipid stores, ER stress and β-cell dysfunction in -challenged RIN-5F cells and isolated pancreatic islets. The lipid-lowering effect of kaempferol was determined by Oil Red O staining, triglyceride assay, BODIPY labeling, RT-PCR and immunoblot analysis of PLIN2 (the lipid droplet coat protein) expression. Further, the involvement of AMPK/mTOR-mediated lipophagy was established by pharmacological and genetic inhibitors of autophagy and AMPK. The co-localization studies of lipid droplets with autophagosomes/lysosomes by BODIPY-MDC-LysoTracker co-staining, LC3/BODIPY labeling and LC3/PLIN2 double immunolabeling further strengthened the findings. Kaempferol treatment exhibited decreased lipid stores and increased co-localization of lipid droplets with autophagosomes and lysosomes in -challenged β-cells. Moreover, inhibition of autophagy led to decreased co-localization and increased lipid droplets accumulation. Kaempferol-induced alleviation of ER stress and β-cell dysfunctions was established by immunoblot analysis of CHOP-10 (a key mediator of cell death in response to ER stress) and insulin content/secretion analysis respectively. Together, these findings suggest that kaempferol prevents ectopic lipid accumulation and ER stress, thus restoring β-cell function through AMPK-mediated lipophagy. The current data implies that kaempferol may be a potential therapeutic candidate to prevent obesity-linked diabetic complications.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Faster Protocol for Endogenous Fatty Esters of Hydroxy Fatty (FAHFA) Measurements.

Fatty esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of endogenous lipids with antidiabetic and anti-inflammatory activities. Interest in these lipids is due to their unique biological activites and the observation that insulin-resistant people have lower esters of hydroxystearic (PAHSA) levels, suggesting that a FAHFA deficiency may contribute to metabolic disease. Rigorous testing of this hypothesis will require the measurement of many clinical samples; however, current analytical workflows are too slow to enable samples to be analyzed quickly. Here we describe the development of a significantly faster workflow to measure FAHFAs that optimizes the fractionation and chromatography of these lipids. We can measure FAHFAs in 30 min with this new protocol versus 90 min using the older protocol with comparable performance in regioisomer detection and quantitation. We also discovered through this optimization that oleic esters of hydroxystearic acids (OAHSAs), another family of FAHFAs, have a much lower background signal than PAHSAs, which makes them easier to measure. Our faster workflow was able to quantify changes in PAHSAs and OAHSAs in mouse tissues and human plasma, highlighting the potential of this protocol for basic and clinical applications.

Keyword: diabetes

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of , obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe liver inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: diabetes

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high fat diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, fatty acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and lipid homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and liver, gastrocnemius and brown fat of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: diabetes

Myo-inositol alters 13C-labeled fatty metabolism in human placental explants.

We postulate that myo-inositol, a proposed intervention for gestational-, affects transplacental lipid supply to the fetus. We investigated the effect of myo-inositol on fatty- processing in human placental-explants from uncomplicated pregnancies. Explants were incubated with 13C-labeled , 13C-oleic- and 13C-docosahexaenoic- across a range of myo-inositol concentrations for 24 h and 48 h. The incorporation of labeled-fatty-acids into individual lipids was quantified by liquid-chromatography-mass-spectrometry. At 24 h, myo-inositol increased the amount of 13C- and 13C-oleic- labeled lipids (median fold-change relative to control=1). Significant effects were seen with 30 µM myo-inositol (physiological) for 13C--lysophosphatidylcholines (1.26) and 13C--phosphatidylethanolamines (1.17). At 48 h, myo-inositol addition increased 13C-oleic--lipids but decreased 13C- and 13C-docosahexaenoic- lipids. Significant effects were seen with 30 µM myo-inositol for 13C-oleic--phosphatidylcholines (1.25), 13C-oleic--phosphatidylethanolamines (1.37) and 13C-oleic--triacylglycerols (1.32) and with 100 µM myo-inositol for 13C-docosahexaenoic--triacylglycerols (0.78). Lipids labeled with the same 13C-fatty- showed similar responses when tested at the same time-point, suggesting myo-inositol alters upstream processes such as fatty- uptake or activation. Myo-inositol supplementation may alter placental lipid physiology with unknown clinical consequences.

Keyword: diabetes

Interesterified palm olein lowers postprandial glucose-dependent insulinotropic polypeptide response in type 2 .

We aim to investigate the postprandial effects of palm olein (PO) and chemically interesterified palm olein (IPO) with different proportions of at the sn-2 position using high oleic sunflower oil (HOS) as control fat on concentrations of gut hormones, glucose homeostasis, satiety, lipid and inflammatory parameters in type 2 diabetic (T2D) subjects.Using a randomised double-blind crossover design, 21 (men\u2009=\u20096, women\u2009=\u200915) T2D subjects consumed test meals (3.65\xa0MJ) consisting of a high fat muffin (containing 50\xa0g test fats provided as PO, IPO or HOS) and a milkshake. Postprandial changes in gut hormones, glucose homeostasis, satiety, lipid and inflammatory parameters after meals were analysed. Some of the solid fractions of the IPO were removed and thus the fatty composition of the PO and IPO was not entirely equal (PO vs IPO: palmitate 39.8 vs 38.7; oleate 43.6 vs 45.1). PO, IPO and HOS contained 9.7, 38.9 and 0.2\xa0g/100\xa0g total fatty acids of at the sn-2 position, respectively. At 37\xa0°C, IPO contained 4.2% SFC whereas PO and HOS were completely melted.Our novel observation shows that the incremental area under curve (iAUC) 0-6\xa0h of plasma GIP concentration was on average 16% lower following IPO meal compared with PO and HOS (P\u2009<\u20090.05) meals. Serum C-peptide concentrations exhibited a significant meal\u2009×\u2009gender interaction (P\u2009=\u20090.009). No differences between test meals were noted for other measurements.This study shows no adverse effect of interesterification on hormones associated with glucose homeostasis notably GLP-1 in T2D subjects.ClinicalTrials.gov . https://clinicaltrials.gov/ct2/show/.

Keyword: diabetes

Circulating Very-Long-Chain SFA Concentrations Are Inversely Associated with Incident Type 2 in US Men and Women.

Very-long-chain SFAs (VLCSFAs), such as arachidic (20:0), behenic (22:0), and lignoceric (24:0), have demonstrated inverse associations with cardiometabolic conditions, although more evidence is needed to characterize their relation with risk of type 2 (T2D). In addition, little is known regarding their potential dietary and lifestyle predictors.We aimed to examine the association of plasma and erythrocyte concentrations of VLCSFAs with incident T2D risk.We used existing measurements of fatty concentrations in plasma and erythrocytes among 2854 and 2831 participants in the Nurses\' Health Study (NHS) and Health Professionals Follow-Up Study (HPFS), respectively. VLCSFAs were measured using GLC, and individual fatty concentrations were expressed as a percentage of total fatty acids. Incident T2D cases were identified by self-reports and confirmed by a validated supplementary questionnaire. Cox proportional hazards regression was used to evaluate the association between VLCSFAs and T2D, adjusting for demographic, lifestyle, and dietary variables.During 39,941 person-years of follow-up, we documented 243 cases of T2D. Intakes of peanuts, peanut butter, vegetable fat, dairy fat, and /stearic (16:0-18:0) fatty acids were significantly, albeit weakly, correlated with plasma and erythrocyte VLCSFA concentrations (|rs|\xa0≤\xa00.19). Comparing the highest with the lowest quartiles of plasma concentrations, pooled HRs (95% CIs) were 0.51 (0.35, 0.75) for arachidic , 0.43 (0.28, 0.64) for behenic , 0.40 (0.27, 0.61) for lignoceric , and 0.41 (0.27, 0.61) for the sum of VLCSFAs, after multivariate adjustments for demographic, lifestyle, and dietary factors. For erythrocyte VLCSFAs, only arachidic and behenic concentrations were inversely associated with T2D risk.Our findings suggest that, in US men and women, higher plasma concentrations of VLCSFAs are associated with lower risk of T2D. More research is needed to understand the mechanistic pathways underlying these associations.Copyright © The Author(s) 2019.

Keyword: diabetes

The monounsaturated fatty oleate is the major physiological toxic free fatty for human beta cells.

Free fatty acids (FFAs) can cause glucose intolerance and . Lipotoxicity to the pancreatic beta cells is considered to be a major underlying cause for this phenomenon. The aim of this study was to analyse the toxicity profile of FFAs in the human EndoC-βH1 beta-cell line and to compare the results with isolated rat and human islets with special reference to the physiologically most prevalent FFAs (PA) and oleic (OA). Toxicity after a 2-day incubation with the different FFAs was analysed by the caspase-3 assay and confirmed by the propidium iodide and annexin V staining tests. The long-chain saturated PA (C16:0) and the monounsaturated OA (C18:1) were both toxic to human EndoC-βH1 beta cells and pseudoislets, as well as to rat islets, and, as confirmed in a pilot experiment, also to human islets. Furthermore, OA provided no protection against the toxicity of PA. Likewise, elaidic (EA, the trans isomer of OA; trans-OA) was significantly toxic, in contrast to the non-metabolisable analogues methylated PA (MePA) and methylated OA (MeOA). Fatty acids with a chain length\u2009\u2009< C16 were not toxic in EndoC-βH1 beta cells. Caspase-3 was also activated by linoleic (LA)(C18:2) but not by γ-linolenic (γ-LNA)(C18:3). Overall, only long-chain FFAs with chain lengths\u2009 >\u2009C14, which generate hydrogen peroxide in the peroxisomal beta-oxidation, were toxic. This conclusion is also supported by the toxicity of the branched-chain FFA pristanic , which is exclusively metabolised in the peroxisomal beta-oxidation. The lack of a protective effect of the monounsaturated fatty OA has important consequences for a beta-cell protective lipid composition of a diet. A cardioprotective diet with a high OA content does not fulfil this requirement.

Keyword: diabetes

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular Inflammation in HepG2 Cells.

Fatty acids induced hepatic inflammation plays an important role in nonalcoholic fatty liver disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti-inflammation in various human organs. However, the anti-inflammation property of HS in the fatty liver is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free fatty acids-induced inflammation. The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

20-HETE promotes glucose-stimulated insulin secretion in an autocrine manner through FFAR1.

The long-chain fatty receptor FFAR1 is highly expressed in pancreatic β-cells. Synthetic FFAR1 agonists can be used as antidiabetic drugs to promote glucose-stimulated insulin secretion (GSIS). However, the physiological role of FFAR1 in β-cells remains poorly understood. Here we show that 20-HETE activates FFAR1 and promotes GSIS via FFAR1 with higher potency and efficacy than dietary fatty acids such as , linoleic, and α-linolenic . Murine and human β-cells produce 20-HETE, and the ω-hydroxylase-mediated formation and release of 20-HETE is strongly stimulated by glucose. Pharmacological inhibition of 20-HETE formation and blockade of FFAR1 in islets inhibits GSIS. In islets from type-2 diabetic humans and mice, glucose-stimulated 20-HETE formation and 20-HETE-dependent stimulation of GSIS are strongly reduced. We show that 20-HETE is an FFAR1 agonist, which functions as an autocrine positive feed-forward regulator of GSIS, and that a reduced glucose-induced 20-HETE formation contributes to inefficient GSIS in type-2 .

Keyword: diabetes

Melatonin improves insulin resistance and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with insulin resistance and type 2 . The aim of this study was to determine whether melatonin improves hepatic insulin resistance and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve insulin resistance in FETUA-overexpressing cells, indicating that improvement in insulin resistance by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced body weight gain to one-third of that of the HFD group and hepatic steatosis. Insulin sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the liver and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic insulin resistance and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: diabetes

The BACE1 product sAPPβ induces ER stress and inflammation and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and fatty oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, insulin resistance, PGC-1α down-regulation, and fatty oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and insulin resistance in skeletal muscle and gluconeogenesis in liver.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Chronic -induced lipotoxicity correlates with defective trafficking of ATP sensitive potassium channels in pancreatic β cells.

Lipotoxicity is associated with a high level of fatty accumulation in pancreatic β-cells. An overload of free fatty acids contributes to pancreatic β-cell apoptosis and dysfunction. Insulin secretion involves sequential ionic events upon glucose stimulation. ATP sensitive potassium (K) channels serve as glucose sensors and effectively initiate glucose-stimulated insulin secretion. This study investigated the effects of lipotoxicity on the trafficking of K channels in pancreatic β cells using chronic -injected mice and treated insulinoma cells. The chronic -injected mice displayed type II diabetic characteristics. The pancreatic sections of these mice exhibited a decrease in the expression of K channels. We then tested the time and dose effects of on the cell viability of INS-1 cells. We observed a significant decrease in the surface expression of K channels after 72 h of treatment with 0.4 mM . In addition, this treatment induced pancreatic β-cell apoptosis by increasing cleaved caspase 3 protein level. Our results demonstrated cotreatment with glibenclamide, the sulfonylurea compounds for type II , in -treated cells reduces cell death and recovers the glucose stimulated insulin secretion through increasing the surface expression of K channels. Importantly, glibenclamide also improved glucose tolerance, triglyceride concentration, and insulin sensitivity in the -injected mice. In conclusion, an increase in the surface expression of K channels restores insulin secretion, reduces pancreatic β-cell\'s apoptosis, highlighting correct trafficking of K channels is important in survival of β-cells during lipotoxicity.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and .

Danzhi jiangtang capsule (DJC) is mainly used to treat type 2 and vascular complication. Our aim was to investigate whether the protective effects of DJC by oral administration on high-fat diet (HFD) and -induced damages of endothelial cells are related to oxidative stress and endoplasmic reticulum (ER) stress. Male Sprague-Dawley rats were randomly divided into standard chow diet (SCD), HFD, HFD plus DJC-low dose (HFD\u2009+\u2009DJCL) and HFD plus DJC-high dose treatment groups (HFD\u2009+\u2009DJCH). The latter three groups were given HFD feeding for three months, then the HFD\u2009+\u2009DJCL and HFD\u2009+\u2009DJCH rats were treated with DJC (500 and 1000\u2009mg/kg/day) via gavage for another two months respectively. Endothelium-dependent relaxations induced by acetylcholine were observed in isolated rat thoracic aortic ring preparation. Malondialdehyde (MDA), total-antioxidant capacity (T-AOC), superoxide dismutase (SOD), interleukin 1β (IL-1β), tumour necrosis factor α (TNFα), free fatty acids (FFA), endothelin-1 (ET-1) and nitric oxide (NO) levels in serum were assayed. The pathological changes, protein expression of endothelial NO synthase (eNOS), phosphorylated eNOS (p-eNOS) and ER stress-related genes in the thoracic aorta were measured. Human umbilical vein endothelial cells (HUVEC) were treated with serum-medicated DJC and then with to detect the reactive oxygen species (ROS) levels and C/EBP homologous protein (CHOP) distribution, expression of p-eNOS and ER stress-related genes. DJC therapy exhibited a potent antioxidant activity and effectively prevented the endothelial dysfunction (ED) manifested by promoting p-eNOS expression and enhancing NO release, decreasing lipid deposition (Oil-red O staining, CPT1b and ACC) and inflammation (IL-1β, TNFα, CD68 and p-JNK), alleviating oxidative and ER stress, and decreasing the apoptosis of endothelial cells (TUNEL, BCL-2 and BAX) induced by HFD and respectively. These findings suggest that the administration of DJC had endothelial protective effects against HFD-induced ED, inflammation and apoptosis by alleviating lipid deposition, oxidative and ER stress.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: diabetes

Metabolomics Study of Metabolic Changes in Renal Cells in Response to High-Glucose Exposure Based on Liquid or Gas Chromatography Coupled With Mass Spectrometry.

Diabetic nephropathy (DN) is one of the most serious microvascular complications and the leading causes of death in (DM). To find biomarkers for prognosing the occurrence and development of DN has significant clinical value for its prevention, diagnosis, and treatment. In this study, a non-targeted cell metabolomics-based ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry and gas chromatography coupled with mass spectrometry was developed and performed the dynamic metabolic profiles of rat renal cells including renal tubular epithelial cells (NRK-52E) and glomerular mesangial cells (HBZY-1) in response to high glucose at time points of 12\u2009h, 24 h, 36 h, and 48 h. Some potential biomarkers were then verified using clinical plasma samples collected from 55 healthy volunteers, 103 DM patients, and 57 DN patients. Statistical methods, such as principal component analysis and partial least squares to latent structure-discriminant analysis were recruited for data analyses. As a result, and linoleic (all-cis-9,12) were the potential indicators for the occurrence and development of DN, and valine, leucine, and isoleucine could be used as the prospective biomarkers for DM. In addition, rise and fall of leucine and isoleucine levels in plasma could be used for prognosing DN in DM patients. Through this study, we established a novel non-targeted cell dynamic metabolomics platform and identified potential biomarkers that may be applied for the diagnosis and prognosis of DM and DN.

Keyword: diabetes

Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high-fat diet-fed mice.

Type 2 is frequently accompanied by fatty liver disease. Lipid accumulation within the liver is considered as one of the risk factors for insulin resistance. Hepatocyte growth factor (HGF) is used to treat liver dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting insulin and homeostatic model assessment of insulin resistance were calculated for insulin sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/fatty synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic insulin resistance and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Investigation published by Asian Association for the Study of (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: diabetes

Resveratrol attenuates type 2 by mediating mitochondrial biogenesis and lipid metabolism via Sirtuin type 1.

The rising incidence of type 2 (T2DM) is a major public health problem and novel therapeutic strategies are required to prevent and treat T2DM. It has been demonstrated that resveratrol (RSV) may prevent T2DM by targeting Sirtuin type 1 (SIRT1), indicating that SIRT1 may be a novel therapeutic target for T2DM prevention. In the present study, a T2DM rat model was established by administering a high fat diet and streptozotocin (STZ) injections. Measurements of blood glucose and insulin confirmed successful establishment of the T2DM model. RSV was used to treat rats with STZ-induced T2DM and the results indicated that RSV reversed the STZ-induced downregulation of peroxisome proliferator-activated receptor-γ coactivator-1α, SIRT1 and forkhead box protein O 3a. Furthermore, RSV modulated the activity of superoxide dismutase and malondialdehyde, which are associated with oxidative stress. , cells from the insulinoma cell line clone 1E were pretreated with (PA) to simulate a high fat environment. The results of reverse transcription-quantitative polymerase chain reaction indicated that PA suppressed the expression of SIRT1 in a dose- and time-dependent manner. Furthermore, PA modulated the expression of mitochondrial biogenesis-associated, lipid metabolism-associated and β-cell-associated genes, whereas RSV treatment ameliorated the PA-induced changes in the expression of these genes via SIRT1. The results of the present study suggest that RSV participates in the prevention of T2DM by regulating the expression of mitochondrial genes associated with biogenesis, lipid metabolism and β-cells via SIRT1. The results of the current study provide an insight into the mechanisms by which SIRT1 inhibits T2DM and may be used as a basis for future studies.

Keyword: diabetes

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 . Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and lipid metabolism. Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: diabetes

lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑fat diet (HFD)‑fed mice to verify the role of MEG3 in hepatic insulin resistance in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 .

Keyword: diabetes

Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells.

Free fatty is considered to be one of the major pathogenic factors of inducing insulin resistance. The association between iron disturbances and insulin resistance has recently begun to receive a lot of attention. Although skeletal muscles are a major tissue for iron utilization and storage, the role of iron in palmitate (PA)-induced insulin resistance is unknown. We investigated the molecular mechanism underlying iron dysregulation in PA-induced insulin resistance. Interestingly, we found that PA simultaneously increased intracellular iron and induced insulin resistance. The iron chelator deferoxamine dramatically inhibited PA-induced insulin resistance, and iron donors impaired insulin sensitivity by activating JNK. PA up-regulated transferrin receptor 1 (tfR1), an iron uptake protein, which was modulated by iron-responsive element-binding proteins 2. Knockdown of tfR1 and iron-responsive element-binding proteins 2 prevented PA-induced iron uptake and insulin resistance. PA also translocated the tfR1 by stimulating calcium influx, but the calcium chelator, BAPTA-AM, dramatically reduced iron overload by inhibiting tfR1 translocation and ultimately increased insulin sensitivity. Iron overload may play a critical role in PA-induced insulin resistance. Blocking iron overload may thus be a useful strategy for preventing insulin resistance and .-Cui, R., Choi, S.-E., Kim, T. H., Lee, H. J., Lee, S. J., Kang, Y., Jeon, J. Y., Kim, H. J., Lee, K.-W. Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells.

Keyword: diabetes

Circulating sphingolipids, fasting glucose, and impaired fasting glucose: The Strong Heart Family Study.

Animal studies suggest sphingolipids as an early marker of impaired glucose metabolism; however, research in humans is limited. We evaluated whether individual sphingolipid species were associated with fasting plasma glucose and incident impaired fasting glucose in a longitudinal cohort study.We measured 15 sphingolipid species from blood samples collected in 2001-2003 from 2145 participants without prevalent in the Strong Heart Family Study. Fasting plasma glucose was measured in blood samples collected at baseline and follow-up (mean 5.5\u202fyears after baseline).The average age of study participants was 38\u202fyears; 41% were men. Ceramide, sphingomyelin, and glucosylceramide species levels were higher in older participants; lactosyl-ceramide levels were higher in participants with lower BMIs. In adjusted analyses, greater concentrations of most ceramide species and lower lactosyl-ceramide with (LC-16) were associated with higher glucose levels at baseline. We did not observe associations of sphingomyelin species or glucosyl-ceramide species with glucose levels. Associations of sphingolipid levels with fasting glucose levels at follow-up were similar but had greater uncertainty than associations with baseline glucose. Although no statistically significant associations of sphingolipids with incident impaired fasting glucose were present, results were similar to glucose analyses.We identified several ceramide species associated with higher fasting glucose levels and one sphingolipid, LC-16, that was associated with lower fasting glucose levels. These findings compliment previous research, which linked these sphingolipids with fasting insulin levels, and suggest that higher levels of these ceramides and lower LC-16 may be an early marker of impaired glucose metabolism. FUND: US National Institutes Health.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: diabetes

Endogenous Fatty Acids Are Essential Signaling Factors of Pancreatic β-Cells and Insulin Secretion.

The secretion of insulin from β-cells depends on extracellular factors, in particular glucose and other small molecules, some of which act on G-protein-coupled receptors. Fatty acids (FAs) have been discussed as exogenous secretagogues of insulin for decades, especially after the FA receptor GPR40 (G-protein-coupled receptor 40) was discovered. However, the role of FAs as endogenous signaling factors has not been investigated until now. In the present work, we demonstrate that lowering endogenous FA levels in β-cell medium by stringent washing or by the application of FA-free (FAF) BSA immediately reduced glucose-induced oscillations of cytosolic Ca ([Ca] oscillations) in MIN6 cells and mouse primary β-cells, as well as insulin secretion. Mass spectrometry confirmed BSA-mediated removal of FAs, with , stearic, oleic, and elaidic being the most abundant species. [Ca] oscillations in MIN6 cells recovered when BSA was replaced by buffer or as FA levels in the supernatant were restored. This was achieved by recombinant lipase-mediated FA liberation from membrane lipids, by the addition of FA-preloaded FAF-BSA, or by the photolysis of cell-impermeant caged FAs. Our combined data support the hypothesis of FAs as essential endogenous signaling factors for β-cell activity and insulin secretion.© 2018 by the American Association.

Keyword: diabetes

MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue inflammation in obesity. Both MIP-1α and free fatty acids are elevated in obesity/T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling pathways were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling pathways. In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in obesity/T2D may contribute to metabolic inflammation through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of obesity, insulin resistance, type-2 and metabolic syndrome. Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal metabolic disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA metabolism on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: diabetes

The effect of FADS2 gene rs174583 polymorphism on desaturase activities, fatty profile, insulin resistance, biochemical indices, and incidence of type 2 .

In this study, we investigated the associations of erythrocytes fatty composition, activities of delta-5 desaturase (D5D) and delta-6 desaturase (D6D), and other metabolic risk factors, with type 2 (T2D) risk to determine if rs174583 polymorphism of FADS2 gene had any effect on these associations.Fatty profile of erythrocytes was determined using gas chromatography-mass spectrometry in 95 T2D patients and 95 apparently healthy participants. The genotypes of single-nucleotide polymorphism (SNP) of FADS2 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism technique. Other biochemical parameters were measured in the serum using standard analytical procedures.D6D activity was increased ( < 0.001) and D5D activity was decreased in T2D patients ( < 0.001) compared to controls. Homeostatic model assessment insulin resistance (HOMA-IR) index was positively correlated with D6D ( = 0.34, < 0.001) and negatively correlated with D5D ( = -0.19, = 0.02). ( < 0.001) and dihomo-gamma-linolenic ( = 0.03) were higher and linoleic ( < 0.001) and arachidonic (AA) ( < 0.001) were lower in T2D patients. The distribution of rs174583 genotypes which includes C/T, C/C, and T/T was not different in the two groups ( = 0.63).In the population studied, there was a strong association in the erythrocytes fatty composition, D5D and D6D activities and other metabolic risk factors between non-T2D and T2D patients. In addition, there was a strong association in erythrocytes DGLA and AA contents and D5D activities between rs174583 genotypes in all participants. However, the distribution of rs174583 genotypes did not differ significantly between T2D patient and controls, and it did not appear to be an association between rs174583 SNP and incident of T2D.

Keyword: diabetes

Induces Müller Cell Inflammation that is Potentiated by Co-treatment with Glucose.

Chronic hyperglycemia is thought to be the major stimulator of retinal dysfunction in diabetic retinopathy (DR). Thus, many -related systemic factors have been overlooked as inducers of DR pathology. Cell culture models of retinal cell types are frequently used to mechanistically study DR, but appropriate stimulators of DR-like factors are difficult to identify. Furthermore, elevated glucose, a gold standard for cell culture treatments, yields little to no response from many primary human retinal cells. Thus, the goal of this project was to demonstrate the effectiveness of the free fatty , and compare its use alone and in combination with elevated glucose as a stimulus for human Müller cells, a retinal glial cell type that is activated early in DR pathogenesis and uniquely responsive to fatty acids. Using RNA sequencing, we identified a variety of DR-relevant pathways, including NFκB signaling and inflammation, intracellular lipid signaling, angiogenesis, and MAPK signaling, that were stimulated by , while elevated glucose alone did not significantly alter any -relevant pathways. Co-treatment of high glucose with potentiated the expression of several DR-relevant angiogenic and inflammatory targets, including PTGS2 (COX-2) and CXCL8 (IL-8).

Keyword: diabetes

Crucial Roles of 5-HT and 5-HT2 Receptor in -Related Lipid Accumulation and Pro-Inflammatory Cytokine Generation in Hepatocytes.

Previously, we confirmed that liver-synthesized 5-HT rather than non-liver 5-HT, acting on the 5-HT2 receptor (5-HT2R), modulates lipid-induced excessive lipid synthesis (ELS). Here, we further revealed the effects of the hepatocellular 5-HT system in -related disorders.Studies were conducted in male ICR mice, human HepG2 cells, and primary mouse hepatocytes (PMHs) under gene or chemical inhibition of the 5-HT system, key lipid metabolism, and inflammation-related factors. Protein and messenger RNA expression and levels of the factors were determined via western blotting, reverse transcription PCR, and quantitative assay kits, respectively. Hepatic steatosis with inflammation and fibrosis, intracellular lipid droplet accumulation (LDA), and reactive oxygen species (ROS) location were determined via hematoxylin and eosin, Masson\'s trichrome, Oil red O, and fluorescent-specific staining, respectively. induced the activation of the 5-HT system: the activation of 5-HT2R, primarily 5-HT2AR, in addition to upregulating monoamine oxidase A (MAO-A) expression and 5-HT synthesis, by activating the G protein/ phospholipase C pathway modulated PKCε activation, resulting in ELS with LDA; the activation of NF-κB, which mediates the generation of pro-inflammatory cytokines, was primarily due to ROS generation in the mitochondria induced by MAO-A-catalyzed 5-HT degradation, and secondarily due to the activation of PKCε. These effects of the 5-HT system were also detected in - or high glucose-treated PMHs and regulated multiple inflammatory signaling pathways. In diabetic mice, co-treatment with antagonists of both 5-HT synthesis and 5-HT2R significantly abolished hepatic steatosis, inflammation, and fibrosis as well as hyperglycemia and dyslipidemia.Activation of the hepatocellular 5-HT system plays a crucial role in inducing -related hepatic dysfunction and is a potential therapeutic target.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Identification of early biological changes in palmitate-treated isolated human islets.

Long-term exposure to elevated levels of free fatty acids (FFAs) is deleterious for beta-cell function and may contribute to development of type 2 (T2DM). Whereas mechanisms of impaired glucose-stimulated insulin secretion (GSIS) in FFA-treated beta-cells have been intensively studied, biological events preceding the secretory failure, when GSIS is accentuated, are poorly investigated. To identify these early events, we performed genome-wide analysis of gene expression in isolated human islets exposed to fatty palmitate for different time periods.Palmitate-treated human islets showed decline in beta-cell function starting from day two. Affymetrix Human Transcriptome Array 2.0 identified 903 differentially expressed genes (DEGs). Mapping of the genes onto pathways using KEGG pathway enrichment analysis predicted four islet biology-related pathways enriched prior but not after the decline of islet function and three pathways enriched both prior and after the decline of islet function. DEGs from these pathways were analyzed at the transcript level. The results propose that in palmitate-treated human islets, at early time points, protective events, including up-regulation of metallothioneins, tRNA synthetases and fatty -metabolising proteins, dominate over deleterious events, including inhibition of fatty detoxification enzymes, which contributes to the enhanced GSIS. After prolonged exposure of islets to palmitate, the protective events are outweighed by the deleterious events, which leads to impaired GSIS.The study identifies temporal order between different cellular events, which either promote or protect from beta-cell failure. The sequence of these events should be considered when developing strategies for prevention and treatment of the disease.

Keyword: diabetes

Oxidative stress induced by modulates K2.3 channels in vascular endothelium.

Elevated plasma free fatty acids level has been implicated in the development of insulin resistance, inflammation, and endothelial dysfunction in diabetic and nondiabetic individuals. However, the underlying mechanisms still remain to be defined. Herein, we investigated the effect of (PA), the most abundant saturated fatty in the human body, on small-conductance Ca-activated potassium channels (K2.3)-mediated relaxation in rodent resistance arteries and the underlying molecular mechanism. The effect of PA on K2.3 in endothelium was evaluated using real-time PCR, Western blotting, whole-cell patch voltage-clamp, wire and pressure myograph system, and reactive oxygen species (ROS) were measured by using dihydroethidium and 2\', 7\'-dichlorofluorescein diacetate. K2.3-mediated vasodilatation responses to acetylcholine and NS309 (agonist of K2.3 and K3.1) were impaired by incubation of normal mesenteric arteries with 100\u202fμM\u202fPA for 24\u202fh. In cultured human umbilical vein endothelial cells (HUVECs), PA decreased K2.3 current and expression at mRNA and protein levels. Incubation with the NADPH oxidase (Nox) inhibitor dibenziodolium (DPI) partly inhibited the PA-induced ROS production and restored K2.3 expression. Inhibition of either p38-MAPK or NF-κB using specific inhibitors (SB203580, SB202190 or Bay11-7082, pyrrolidinedithiocarbamate) attenuated PA-induced downregulation of K2.3 and inhibition of p38-MAPK also attenuated PA-induced phosphorylation of NF-κB p65. Furthermore, DPI reversed the increment of phospho-p38-MAPK by PA. These results demonstrated that PA downregulated K2.3 expressions via Nox/ROS/p38-MAPK/NF-κB signaling leading to endothelial vasodilatory dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro research.

The interplay between hyper-glycemia and -lipidemia in (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for lipid transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Adiponectin homolog novel osmotin protects obesity/-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated .To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Time-Dependent alteration to the tight junction structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal microbiota composition, increased intestinal permeability, and onset of type 2 (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the tight junction (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the microbiota, after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Epigallocatechin gallate improves insulin resistance in HepG2 cells through alleviating inflammation and lipotoxicity.

High levels of circulating free fatty acids (FFAs), inflammation and oxidative stress are important causes for insulin resistance (IR) and type 2 . The aim of this study was to investigate the mechanisms of EGCG in alleviating IR in HepG2 cells.HepG2 cells were treated with 25\u202fmM glucose, 0.25\u202fmM (PA), or 50\u202fμM EGCG for 24\u202fh.EGCG increased glucose uptake and decreased glucose content. EGCG markedly decreased the levels of inflammatory and oxidative stress factors including nuclear factor κB (NF-κB), tumor necrosis factor-α, interleukin-6, reactive oxygen species, malondialdehyde and p53 protein, and markedly increased superoxide dismutases (SOD), glutathione peroxidase and SOD2 protein. EGCG significantly downregulated the levels of FFAs, triacylglycerol and cholesterol in HepG2 cells. The glucose transporter 2 (GLUT2) protein and its downstream proteins peroxisome proliferator-activated receptor γ coactivator (PGC)-1β were significantly increased, and sterol regulatory element-binding-1c (SREBP-1c) protein, and fatty synthase (FAS) were significantly decreased by EGCG in HepG2. Moreover, the foregoing effects were reversed by siRNA-mediated knockdown of GLUT2.Our data demonstrated that EGCG improved IR, possibly through ameliorating glucose (25\u202fmM) and PA (0.25\u202fmM)-induced inflammation, oxidative stress, and FFAs via the GLUT2/PGC-1β/SREBP-1c/FAS pathway in HepG2 cells.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

A novel PPARα/γ agonist, propane-2-sulfonic octadec-9-enyl-amide, ameliorates insulin resistance and gluconeogenesis in vivo and vitro.

Peroxisome proliferator-activated receptor alpha/gamma (PPARα/γ) agonists have emerged as important pharmacological agents for improving insulin action. Propane-2-sulfonic octadec-9-enyl-amide (N15) is a novel PPARα/γ dual agonist synthesized in our laboratory. The present study investigates the efficacy and safety of N15 on insulin resistance regulation in high fat diet (HFD)-and streptozotocin (STZ)-induced diabetic mice and in (PA)-induced HepG2 cells. Our results showed that N15 remarkably ameliorated insulin resistance and dyslipidemia in vivo, as well as rectified the glucose consumption and gluconeogenesis in vitro. Moreover, the glucose-lowering effect of N15 was associated with PPARγ mediated up-regulation of hepatic glucose consumption and down-regulation of gluconeogenesis. Meanwhile, N15 exerted advantageous effects on glucose and lipid metabolism without triggering weight gain and hepatotoxicity in mice. In conclusion, our data demonstrated that by alleviating glucose and lipid abnormalities, N15 could be used as a potential prophylactic and therapeutic agent against type 2 and related metabolic disorders.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Fatty and Lipopolysaccharide Effect on Beta Cells Proteostasis and its Impact on Insulin Secretion.

Metabolic overload by saturated fatty acids (SFA), which comprises β-cell function, and impaired glucose-stimulated insulin secretion are frequently observed in patients suffering from obesity and type 2 . The increase of intracellular Ca triggers insulin granule release, therefore several mechanisms regulate Ca efflux within the β-cells, among others, the plasma membrane Ca-ATPase (PMCA). In this work, we describe that lipotoxicity mediated mainly by the saturated (PA) (16C) is associated with loss of protein homeostasis (proteostasis) and potentially cell viability, a phenomenon that was induced to a lesser extent by stearic (18C), myristic (14C) and lauric (12C) acids. PA was localized on endoplasmic reticulum, activating arms of the unfolded protein response (UPR), as also promoted by lipopolysaccharides (LPS)-endotoxins. In particular, our findings demonstrate an alteration in PMCA1/4 expression caused by PA and LPS which trigger the UPR, affecting not only insulin release and contributing to β-cell mass reduction, but also increasing reactive nitrogen species. Nonetheless, stearic (SA) did not show these effects. Remarkably, the proteolytic degradation of PMCA1/4 prompted by PA and LPS was avoided by the action of monounsaturated fatty acids such as oleic and palmitoleic . Oleic recovered cell viability after treatment with PA/LPS and, more interestingly, relieved endoplasmic reticulum (ER) stress. While palmitoleic improved the insulin release, this fatty seems to have more relevant effects upon the expression of regulatory pumps of intracellular Ca. Therefore, chain length and unsaturation of fatty acids are determinant cues in proteostasis of β-cells and, consequently, on the regulation of calcium and insulin secretion.

Keyword: diabetes

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: diabetes

Palmitate and oleate co-treatment increases myocellular protein content via impaired protein degradation.

Protein balance is a crucial determinant of myocellular size and function. The effects of fatty acids on myocellular protein balance remain controversial. The aim of this study was to determine the direct effects of a mixed-species fatty environment on myocellular protein synthesis and degradation.C2C12 myotubes were cultured in media containing equimolar (250\xa0μM) and oleate (PO) or bovine serum albumin control for ≤72\xa0h. Myocellular protein balance was determined via incorporation (synthesis) or release (degradation) of H-tyrosine after 24, 48, and 72\xa0h of treatment. Expression of major proteolytic genes was measured by reverse transcription polymerase chain reaction.PO significantly increased myocellular protein content at 24, 48, and 72\xa0h. Basal myocellular protein synthesis was unchanged by PO. However, PO significantly decreased basal rate of protein degradation at 24\xa0h and this effect persisted throughout 72\xa0h of treatment. Expression of the proteolytic genes Atrogin-1 (MAFbx), MuRF-1, LC3, and ATG4 B, was reduced during the 72\xa0h PO.A mixed-species fatty environment increases myocellular protein content by decreasing the rate of protein degradation, which may be regulated at the level of gene transcription.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: diabetes

, but not high-glucose, induced myocardial apoptosis is alleviated by N‑acetylcysteine due to attenuated mitochondrial-derived ROS accumulation-induced endoplasmic reticulum stress.

Pharmacological inhibition of reactive oxygen species (ROS) is a potential strategy to prevent -induced cardiac dysfunction. This study was designed to investigate precise effects of antioxidant N‑acetylcysteine (NAC) in alleviating diabetic cardiomyopathy (DCM). Echocardiography and histologic studies were performed 12 weeks after streptozocin injection. Protein levels involved in endoplasmic reticulum stress (ERS) and apoptosis were analyzed by western blotting in diabetic hearts or high-glucose (HG, 30\u2009mM)- and (PA, 300\u2009μM)-cultured neonatal rat cardiomyocytes (NRCMs). ROS generation and structural alterations of mitochondria were also assessed. We report that NAC alleviated -induced cardiac abnormality, including restored ejection fraction (EF %), fraction shortening (FS %), peak E to peak A ratio (E/A) and reduced cardiac hypertrophy and fibrosis. These effects were concomitant with blocked ERS and apoptosis, as evidenced by inactivation of phosphorylated inositol-requiring enzyme-1α (IRE1α)/spliced X-box binding protein 1 (XBP1), phosphorylated protein kinase-like kinase (PERK)/phosphorylated eukaryotic initiation factor 2α (eIF2α) and glucose-regulated protein 78 (GRP78)/activating transcription factor 6 (ATF6α)/C/EBP homologous protein (CHOP) pathways, as well as suppressed Bcl-2-associated X protein (BAX)/B-cell lymphoma-2 (Bcl-2) and cleaved caspase 3 expressions. Mechanistically, PA mediated excessive mitochondrial ROS generation and oxidative stress, which were antagonized by NAC and Mito-TEMPO, a mitochondrial ROS inhibitor. No effects were noted by addition of apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, and NADPH oxidase 4 (NOX 4) and NOX 2 expressions were not altered, indicating that PA-induced ROS generation is independent of NADPH oxidases. Most intriguingly, HG failed to promote ROS production despite its ability to promote ERS and apoptosis in NRCMs. Collectively, these findings indicate that NAC primarily abrogates PA-mediated mitochondrial ROS through ERS and therefore alleviates myocardial apoptosis but has little effect on HG-induced cardiac injury. This uncovers a potential role for NAC in formulating novel cardioprotective strategies in DCM patients.

Keyword: diabetes

The novel NADPH oxidase 4 selective inhibitor GLX7013114 counteracts human islet cell death in vitro.

It has been proposed that pancreatic beta-cell dysfunction in type 2 is promoted by oxidative stress caused by NADPH oxidase (Nox) over-activity. The aim of the present study was to evaluate the efficacy of novel Nox inhibitors as protective agents against cytokine- or high glucose + palmitate-induced human beta-cell death. The Nox2 protein was present mainly in the cytoplasm and was induced by cytokines. Nox4 protein immunoreactivity, with some nuclear accumulation, was observed in human islet cells, and was not affected by islet culture in the presence of cytokines or high glucose + palmitate. Nox inhibitors with partial or no isoform selectivity (DPI, dapsone, GLX351322, and GLX481372) all reduced ROS production of human islet cells exposed to high glucose + palmitate. This was paralleled by improved viability and reduced caspase 3 activation. The Nox1 selective inhibitor ML171 failed to reduce human islet cell death in response to both cytokines and high glucose + palmitate. The selective Nox2 inhibitor Phox-I2 also failed to protect against cytokines, but protected partially against high glucose + palmitate-induced cellular death. The highly selective Nox4 inhibitor GLX7013114 protected islet cells against both cytokines and high glucose + palmitate. However, as no osmotic control for high glucose was used, we cannot exclude the possibility that the high glucose effect was due to osmosis. It is concluded that Nox4 may participate in stress-induced islet cell death in human islets in vitro. We propose that Nox4 mediates pro-apoptotic effects in intact islets under stressful conditions and that selective Nox4-inhibition may be a therapeutic strategy in type 2 .

Keyword: diabetes

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only energy metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating obesity-induced inflammation.

Keyword: diabetes

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: dysbiosis

Gut and Metabolome Response of Seed Oil on Metabolism Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.

Keyword: dysbiosis

Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut , and hepatic metabolism.

Keyword: dysbiosis

Vaginal lipidomics of women with vulvovaginal candidiasis and cytolytic vaginosis: A non-targeted LC-MS pilot study.

To characterize the lipid profile in vaginal discharge of women with vulvovaginal candidiasis, cytolytic vaginosis, or no vaginal infection or dysbiosis.Cross-sectional study.Genital Infections Ambulatory, Department of Tocogynecology, University of Campinas, Campinas, São Paulo-Brazil.Twenty-four women were included in this study: eight with vulvovaginal candidiasis, eight with cytolytic vaginosis and eight with no vaginal infections or dysbiosis (control group).The lipid profile in vaginal discharge of the different study groups was determined by liquid chromatography-mass spectrometry and further analyzed with MetaboAnalyst 3.0 platform.Vaginal lipids concentration and its correlation with vulvovaginal candidiasis and cytolytic vaginosis.PCA, PLS-DA and hierarchical clustering analyses indicated 38 potential lipid biomarkers for the different groups, correlating with oxidative stress, inflammation, apoptosis and integrity of the vaginal epithelial tissue. Among these, greater concentrations were found for Glycochenodeoxycholic -7-sulfate, O-adipoylcarnitine, 1-eicosyl-2-heptadecanoyl-glycero-3-phosphoserine, undecanoic , formyl dodecanoate and lipoic in the vulvovaginal candidiasis group; N-(tetradecanoyl)-sphinganine, DL-PPMP, 1-oleoyl-cyclic phosphatidic, and 5-aminopentanoic in the cytolytic vaginosis group; and 1-nonadecanoyl-glycero-3-phosphate, eicosadienoic , 1-stearoyl-cyclic-phosphatidic , 1-(9Z,12Z-heptadecadienoyl)-glycero-3-phosphate, formyl 9Z-tetradecenoate and 7Z,10Z-hexadecadienoic in the control group.Lipids related to oxidative stress and apoptosis were found in higher concentrations in women with vulvovaginal candidiasis and cytolytic vaginosis, while lipids related to epithelial tissue integrity were more pronounced in the control group. Furthermore, in women with cytolytic vaginosis, we observed higher concentrations of lipids related to bacterial overgrowth.

Keyword: dysbiosis

Dynamic alterations in the gut and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut and metabolome.

Keyword: dysbiosis

Effects of eicosapentaenoic and gamma-linolenic acids (dietary lipids) on pulmonary surfactant composition and function during porcine .

To investigate whether a diet enriched with fish and borage oils, with their high polyunsaturated fatty (PUFA) content, alters surfactant composition and function during .Prospective, randomized, blinded, controlled animal study.Research laboratory at a medical center.Thirty-six 15- to 25-kg, disease-free, castrated male pigs. DIETS AND MEASUREMENTS: Three groups of pigs (n = 12 per group) were fed for 8 days diets containing either omega-6 fatty acids (FAs) (corn oil; diet A), or omega-3 FAs (fish oil; diet B), or a combination of omega-6 and omega-3 FAs (borage and fish oils; diet C). Eight of 12 pigs in each group received a 0.1-mg/kg bolus of Escherichia coli endotoxin followed by a continuous infusion (0. 075 mg/kg/h). One lung was subsequently isolated ex vivo, and pressure-volume curves were measured. The contralateral lung was lavaged, and surfactant was analyzed for total and individual phospholipids and FA composition. Minimum and maximum surface tension was measured by bubble surfactometry.Pigs fed either diet B or C had increased oleic (C(18:1) omega-9), eicosapentaenoic (EPA; C(20:5) omega-3), docosahexaenoic (C(22:6) omega-3), and total omega-3 and monounsaturated FAs in their surfactant PUFA pools. The relative percentage of linoleic (C(18:2) omega-6) and total omega-6 FAs were significantly lower from pigs fed diets B and C compared with diet A. (C(16:0)) concentrations, the primary FA in surfactant, had a tendency to be lower in pigs fed diets B and C. There were no demonstrable effects on surfactant function or pulmonary compliance.Diets containing EPA or EPA and gamma-linolenic altered the PUFA composition of pulmonary surfactant, but without demonstrable effects on surfactant function during porcine .

Keyword: endotoximia

Deletion of apoptosis signal-regulating kinase 1 (ASK1) protects pancreatic beta-cells from stress-induced death but not from glucose homeostasis alterations under pro-inflammatory conditions.

Type 2 diabetes is characterized by pancreatic beta-cell dysfunction and is associated with low-grade . Recent observations suggest that apoptosis signal-regulating kinase 1 (ASK1) is involved in beta-cell death in response to different stressors. In this study, we tested whether ASK1 deficiency protects beta-cells from glucolipotoxic conditions and cytokines treatment or from glucose homeostasis alteration induced by endotoxemia.Insulin secretion was neither affected upon shRNA-mediated downregulation of ASK1 in MIN6 cells nor in islets from ASK1-deficient mice. ASK1 silencing in MIN6 cells and deletion in islets did not prevent the deleterious effect of glucolipotoxic conditions or cytokines on insulin secretion. However, it protected MIN6 cells from death induced by ER stress or palmitate and islets from short term caspase activation in response to cytokines. Moreover, endotoxemia induced by LPS infusion increased insulin secretion during hyperglycemic clamps but the response was similar in wild-type and ASK1-deficient mice. Finally, insulin sensitivity in the presence of LPS was not affected by ASK1-deficiency.Our study demonstrates that ASK1 is not involved in beta-cell function and dysfunction but controls stress-induced beta-cell death.

Keyword: endotoximia

Effects of endotoxin infusion on circulating levels of eicosanoids, progesterone, cortisol, glucose and lactic , and abortion in pregnant cows.

The effects of Escherichia coli endotoxin infusions (1.0 or 2.5 micrograms kg-1 over 6 h) on pregnancy were investigated in cows in the first, second and third trimester of gestation. Endotoxin increased the plasma levels of prostaglandins (PGs), thromboxane B2 and cortisol, and decreased progesterone. The severity of the clinical signs and the magnitude of the increases in plasma PGs, thromboxane B2 and cortisol tended to depend on the dose of endotoxin, but were independent of the gestation period. There was hyperglycemia followed by hypoglycemia and lactic acidemia. Hyperglycemia and lactic acidemia were significant only at the high dose of endotoxin. Endotoxin infusion at both doses caused a preferential mobilization of oleic from adipose tissue, and also had some effects on the mobilization of and stearic acids during the post-infusion period. The cows in the first trimester of gestation were more sensitive to the abortifacient effect of endotoxin than cows in the second and third trimester of gestation. The results of this study indicate that the mechanism of endotoxin-induced abortion in cows initially involves a prolonged release of PGF2 alpha and its subsequent stimulant effect on uterine smooth muscle contraction and luteolytic effect leading to a gradual decline in the plasma levels of progesterone. It was concluded that pregnancy terminates in the absence of an adequate level of progesterone, especially during the first trimester of gestation, when progesterone of extraluteal origin is not yet available, coupled with the PGF2 alpha-induced propulsive contraction of the uterus. In addition, the metabolic and circulatory failures in severe cases of , especially at the high dose of endotoxin, resulting either directly or indirectly via the release of various autacoids, catecholamines and cortisol, may also contribute to the termination of pregnancy at any stage of gestation.

Keyword: endotoximia

Potential therapeutic efficacy of inhibitors of human phospholipase A2 in septic shock.

Soluble phospholipase A2 has been implicated in the pathogenesis of local and systemic inflammatory reactions. Elevated levels of circulating phospholipase A2 (PLA2) correlate with the severity of circulatory collapse and pulmonary dysfunction in gram-negative septic shock. Characterization of septic shock serum PLA2 revealed a calcium-dependent enzyme with absolute 2-acyl specificity with a pH optimum of 7.5. We tested a number of therapeutic agents for their ability to inhibit PLA2 from human septic shock serum. Chloroquine, chlorpromazine, dexamethasone base, dexamethasone sodium phosphate, indomethacin, lidocaine, oleic , , promethazine, trans-retinoic , rutin and dl-alpha-tocopherol were all studied over the range of 10(-2) to 10(-7) M. All agents, with the sole exception of dexamethasone base, inhibited PLA2 activity at concentrations greater than 10(-3) M. PLA2 inhibition by dexamethasone sodium phosphate was factitious, due to the formation of calcium-phosphate complexes. Of the 11 agents studied, chlorpromazine was the most effective, with an IC50 of 7.5 X 10(-5) M, a membrane concentration achievable within its therapeutic range. Inhibition was non-competitive with an apparent Ki of 5 nM. Since serum PLA2 levels correlate with mortality in both experimental and clinical gram-negative septic shock, and chlorpromazine was previously shown to improve survival in these conditions, we postulate that its therapeutic efficacy resides at least in part in its PLA2-inhibitory activity.

Keyword: endotoximia

[Synthesis of 4-palmitoyl-sinomenine and its anti- activity].

To synthesize the derivatives of sinomenine, 4-palmitoyl-sinomenine, and explore its therapeutic effect on lipopolysaccharide (LPS)-induced endotoxemia.A highly efficient synthesis of sinomenine derivatives called 4-palmitoyl-sinomenine was made with a molecule of substitutions at C-16 position of ring A. One hour before endotoxemia induction by i.p. injection of 10 mg/kg LPS, high-dose treatment mice (n=5/group) received an i.p. injection of 5 mg/kg sinomenine or 4-palmitoyl-sinomenine while the low-dose treatment mice (n=5/group) received 2.5 mg/kg sinomenine or 4-palmitoyl-sinomenine. Untreated group and normal control group received normal saline. And their survival was monitored hourly for 24 hours. Examination of cytotoxicity of 4-palmitoyl-sinomenine on RAW264.7 cells was conducted at a concentration range of 1 to 125 μmol/L using MTT assay. RAW264.7 cells were exposed to 4-palmitoyl-sinomenine (0, 1, 2, 5, 10 μmol/L) for 24 hours, and then treated with LPS (1 μg/mL) for 6 hours. Then RAW264.7 cells were collected and the mRNA level of IL-6 was detected by real-time quantitative PCR(qRT-PCR).Sinomenine derivatives were successfully synthesized to get 4-palmitoyl-sinomenine. The survival percentage of 4-palmitoyl-sinomenine treatment groups was higher than that of sinomenine treatment groups at the same treatment concentration. The 4-palmitoyl-sinomenine inhibited RAW264.7 cell proliferation and IL-6 gene transcription.The 4-palmitoyl-sinomenine has an anti- probably through inhibiting the proliferation of RAW264.7 cells and decreasing the inflammatory gene expression and inflammatory cytokine release.

Keyword: endotoximia

Effects of smooth and rough Pasteurella haemolytica lipopolysaccharides on plasma cyclic-nucleotides and free fatty acids in calves.

The present study examined the potency of smooth or rough Pasteurella haemolytica lipopolysaccharide infusion (LPS, 24 ng kg-1 min-1 for 500 min) on plasma cyclic-nucleotides and several free fatty acids (FFA) in calves. Both smooth or rough LPS increased plasma cAMP immediately to its maximum at 1 h of infusion, whereas plasma cGMP levels rose slowly and peaked 12 h later. The increases in cAMP levels were more prolonged for smooth LPS than rough LPS. The maximum plasma cAMP rise coincided with increases of several plasma FFA. Rough LPS increased plasma oleic greater than greater than stearic greater than linoleic acids in the second hour and reached their steady state levels between 2 h of infusion and 5 h post-infusion. Thereafter, oleic remained maximally elevated, while stearic decreased and other FFA returned to baseline. Smooth LPS had no effects on and stearic acids, but elevated oleic in an essentially similar manner to rough LPS and increased linoleic initially at 5 h, followed by decreases throughout post-infusion. These results demonstrate that produces early marked elevations in plasma cAMP, a gradual rise in plasma cGMP and disproportionate increases in several plasma FFA. The data also demonstrate that smooth and rough LPS differ in their abilities to increase plasma cAMP and FFA and these may be attributed to differences in their in vivo mechanisms of action. The study suggests that cAMP and cGMP may mediate actions of endotoxin at the cellular level and that differences exist in release and/or utilization of each FFA at different stages of .

Keyword: endotoximia

Anti-inflammatory activity of methyl palmitate and ethyl palmitate in different experimental rat models.

Methyl palmitate (MP) and ethyl palmitate (EP) are naturally occurring fatty esters reported as inflammatory cell inhibitors. In the current study, the potential anti-inflammatory activity of MP and EP was evaluated in different experimental rat models. Results showed that MP and EP caused reduction of carrageenan-induced rat paw edema in addition to diminishing prostaglandin E2 (PGE2) level in the inflammatory exudates. In lipopolysaccharide (LPS)-induced in rats, MP and EP reduced plasma levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). MP and EP decreased NF-κB expression in liver and lung tissues and ameliorated histopathological changes caused by LPS. Topical application of MP and EP reduced ear edema induced by croton oil in rats. In the same animal model, MP and EP reduced neutrophil infiltration, as indicated by decreased myeloperoxidase (MPO) activity. In conclusion, this study demonstrates the effectiveness of MP and EP in combating inflammation in several experimental models.Copyright © 2012 Elsevier Inc. All rights reserved.

Keyword: endotoximia

In vitro Modulation of the LPS-Induced Proinflammatory Profile of Hepatocytes and Macrophages- Approaches for Intervention in Obesity?

Low grade endotoxemia is a feature of obesity which is linked to development of steatohepatitis in non-alcoholic fatty liver disease. In this study, macrophages (J774) and hepatocytes (HepG2) were stimulated with lipopolysaccharide (LPS) from E. coli 0111: B4 and analyzed for modulation of this response when preconditioned or stimulated subsequent to LPS, with different doses of Vitamin D3 or docosahexaenoic (DHA) over a time period of 1 and 5 days. Pro-inflammatory TNFα and pro-fibrotic TGFβ released into the supernatants were measured by ELISA; qPCR was performed for Srebp-1c and PPARα mRNA (genes for products involved in fatty synthesis and catabolism, respectively). Vitamin D3 and DHA exerted a consistent, dose dependent anti-inflammatory effect, and increased PPARα relative to Srebp-1c in both cell types. By contrast, addition of free fatty acids (FFA, oleic / 2:1) caused aggravation of LPS-induced inflammatory reaction and an increase of Srebp-1c relative to PPARα. Our results argue in favor of dietary supplementation of Vitamin D3 or DHA (and avoidance of monounsaturated/saturated fatty acids) to alleviate development of fatty liver disease.

Keyword: endotoximia

Saturated fatty combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic , a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: endotoximia

Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide.

We have previously reported that Smad6, one of the inhibitory Smads of transforming growth factor-β (TGF-β)/bone morphogenetic protein (BMP) signaling, inhibits Toll-like receptor (TLR) 4 signaling by disrupting the Pellino-1-mediated TLR4 signaling complex. Here, we developed Smaducin-6, a novel membrane-tethered -conjugated Smad6-derived peptide composed of amino acids 422-441 of Smad6. Smaducin-6 interacted with Pellino-1, located in the inner membrane, thereby disrupting the formation of IRAK1-, RIP1-, IKKε-mediated TLR4 signaling complexes. Systemic administration of Smaducin-6 showed a significant therapeutic effect on mouse TLR4-mediated inflammatory disease models, cecal-ligation-puncture (CLP)-induced sepsis, and lipopolysaccharide-induced endotoxemia, by inhibiting pro-inflammatory cytokine production and apoptosis while enhancing neutrophil migration and bacterial clearance. Our findings provide clues to develop new peptide-based drugs to target Pellino-1 protein in TLR4 signaling pathway for the treatment of sepsis.© 2015 The Authors. Published under the terms of the CC BY 4.0 license.

Keyword: endotoximia

An insight into nutritional profile of selected Pleurotus species.

The global demand for good quality food indicates that consumers are more concerned about a particular diet associated with good health and lower risk for certain ailments. Mushrooms are widely used as healthy nutritious food. In the present study, the nutritional composition of four different Pleurotus sp. was determined. Prior to extraction, all the selected mushrooms were subjected for proximate composition analysis. The protein, fat, ash, total carbohydrate, fiber and contents were in the range of (16.07- 25.15%), (0.64-2.02%), (2.1-9.14%), (65.66-82.47%), (6.21-54.12%) and (342.20-394.30Kcal/100g), respectively. The spectrophotometric analysis showed that the concentration of protein was in the range of 45.78-33.47 mg/g in all Pleurotus sp. High performance liquid chromatographic analysis of sugars showed six different mono and disaccharides in all the selected mushrooms. The fatty profile by gas chromatography-mass spectrometry revealed that the main fatty acids in selected mushrooms were present in the order linoliec > oleic > . The results suggested that all the Pleurotus sp. could be considered as a rich source of nutrients.

Keyword: energy

Long-term supplementation interacts with parity in lactating dairy cows: Production responses, nutrient digestibility, and partitioning.

The objective of our study was to evaluate the effects of long-term (C16:0) supplementation and parity on production, nutrient digestibility, and partitioning of mid-lactation dairy cows. Forty mid-lactation Holstein cows (18 primiparous and 22 multiparous) were used in a block design. Cows were assigned to receive either a control diet containing no supplemental fat (CON) or a C16:0-enriched supplemented diet (PA; 1.5% diet dry matter) fed for 10 wk. Compared with CON, PA increased dry matter intake, milk yield, cumulative milk yield, milk fat content, milk fat yield, 16-carbon milk fatty (FA) yield, 3.5% fat-corrected milk yield, and -corrected milk yield. Additionally, PA increased body weight change, but did not affect body condition score change compared with CON. A tendency for a treatment by parity interaction was observed for milk yield due to PA increasing milk yield in multiparous but not in primiparous cows. In addition, we observed interactions between treatment and parity for fat-corrected milk, -corrected milk, and milk fat yield due to PA increasing these variables to a greater extent in multiparous compared with primiparous cows. Interestingly, we observed an interaction between treatment and parity for body weight change, due to PA increasing body weight change in primiparous but not in multiparous cows. The PA treatment increased dry matter and neutral detergent fiber digestibilities compared with CON. Although PA did not affect 18-carbon FA digestibility, compared with CON, PA decreased 16-carbon and total FA digestibilities and increased total FA intake by 470 g/d and absorbed total FA by 316 g/d. We also observed an interaction between treatment and parity for total absorbed FA due to PA increasing it to a greater extent in multiparous than in primiparous cows. Compared with CON, PA increased apparent intake and milk output. We observed an interaction between treatment and parity for milk output due to PA increasing milk output to a greater extent in multiparous than primiparous cows. Additionally, an interaction between treatment and parity was observed for output in body reserves due to PA increasing output in body reserves in primiparous but not in multiparous cows. In conclusion, production responses of dairy cows to PA were consistent throughout the 10-wk treatment period. In addition, PA supplementation interacted with parity, with production responses increased to a greater extent in multiparous than primiparous cows and partitioned to body reserves only increased in primiparous cows.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

A novel physiological role for cardiac myoglobin in lipid metabolism.

Continuous contractile activity of the heart is essential and the required is mostly provided by fatty (FA) oxidation. Myocardial lipid accumulation can lead to pathological responses, however the underlying mechanisms remain elusive. The role of myoglobin in dioxygen binding in cardiomyocytes and oxidative skeletal muscle has widely been appreciated. Our recent work established myoglobin as a protector of cardiac function in hypoxia and disease states. We here unravel a novel role of cardiac myoglobin in governing FA metabolism to ensure the physiological production through β-oxidation, preventing myocardial lipid accumulation and preserving cardiac functions. In vivoH magnetic resonance spectroscopy unveils a 3-fold higher deposition of lipids in mouse hearts lacking myoglobin, which was associated with depressed cardiac function compared to wild-type hearts as assessed by echocardiography. Mass spectrometry reveals a marked increase in tissue triglycerides with preferential incorporation of and oleic acids. Phospholipid levels as well as the metabolome, transcriptome and proteome related to FA metabolism tend to be unaffected by myoglobin ablation. Our results reveal a physiological role of myoglobin in FA metabolism with the lipid accumulation-suppressing effects of myoglobin preventing cardiac lipotoxicity.

Keyword: energy

Two Cockayne Syndrome patients with a novel splice site mutation - clinical and metabolic analyses.

Cockayne Syndrome (CS) is a rare autosomal recessive disorder, which leads to neurodegeneration, growth failure and premature aging. Most of the cases are due to mutations in the ERCC6 gene, which encodes the protein CSB. CSB is involved in several functions including DNA repair and transcription. Here we describe two Danish brothers with CS. Both patients carried a novel splice site mutation (c.2382+2T>G), and a previously described nonsense mutation (c.3259C>T, p.Arg1087X) in a biallelic state. Both patients presented the cardinal features of the disease including microcephaly, congenital cataract and postnatal growth failure. In addition, their fibroblasts were hypersensitive to UV irradiation and exhibited increased superoxide levels in comparison to fibroblasts from healthy age and gender matched individuals. Metabolomic analysis revealed a distinctive metabolic profile in cells from the CS patients compared to control cells. Among others, α-ketoglutarate, hydroxyglutarate and certain amino acids (ornithine, proline and glycine) were reduced in the CS patient fibroblasts, whereas glycolytic intermediates (glucose-6-phosphate and pyruvic ) and fatty acids (, stearic and myristic ) were increased. Our data not only provide additional information to the database of CS mutations, but also point towards targets for potential treatment of this devastating disease.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Potential of oleaginous yeast Trichosporon sp., for conversion of sugarcane bagasse hydrolysate into biodiesel.

This study reports production of microbial oil from a yeast strain Trichosporon sp., (RW) isolated from decayed wood. Preliminary analysis based on fluorescence microscopy and spectroscopy of Nile red stained yeast cells showed accumulation of lipid globules. The potential of the yeast to produce lipids was evaluated on glucose, glycerol and hydrolysate of sugarcane bagasse, where Trichosporon sp. (RW) was found to accumulate 21.45 (59.6%), 18.41 (56%) and 10.25g/l (40.5%) of the lipids after 120h of fermentation at 30°C. FAME analysis of lipids by GC-FID and NMR revealed oleic (18:1) as the major constituent, corresponding to 50.05, 46.48 and 54.66% of the accumulated lipids in glucose, glycerol and hydrolysate grown cultures, respectively. Other accumulated lipids included (16:0), linoleic (18:2) and stearic acids (18:0) in that order. The cetane number of the lipids ranged from 52.39 to 59.57 indicating suitability for biodiesel production.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

[The becoming of reactions of lipolysis in phylogenesis. The and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V].

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring. In lipoproteins of very low density and lipoproteins of low density, the carnivorous transfer exogenous non-polar triglycerides. The herbivorous also transfer non-polar triglycerides though synthesized by hepatocytes from glucose endogenically. In herbivorous, transferring of non-polar triglycerides prior to synthesis of insulin is forming apoB-100 in composition of lipoproteins of very low density and lipoproteins of low density. The hydrolysis of non-polar triglycerides in lipoproteins of very low density is activated by hepatic glycerol hydrogenase and apoC-III; cells absorb lipoproteins of low density by means of apoB-100 endocytosis. The content on lipoproteins in blood plasma under electrophoresis of lipoproteins corresponds to hepatic glycerol hydrogenase type IIb. In frst and second types of fatty acids transferring in form of triglycerides to lipoproteins of very low density + lipoproteins of low density predominate fatty , triglycerides of the same name and metabolism of fatty acids in vivo. The insulin initiated the third type of transferring of oleic fatty by now to insulin-depended cells only in oleic lipoproteins of very low density; hydrolysis of oleic triglycerides is activated by late in phylogenesis post-heparin hepatic glycerol hydrogenase and apoC-II cofactor. The dynamic apoE is actively bound by apoB-100 forming apoE/B-100 ligand. At later stages of phylogenesis insulin formed fatty acids transferring in form of oleic triglycerides in lipoproteins of very low density of the same name without forming of oleic lipoproteins of low density; the electrophoregram of lipoproteins reflects absence of hepatic glycerol hydrogenase. In phylogenesis three types of fatty acids transferring to triglycerides in composition of lipoproteins formed sequentially: 1) chylomicrons + lipoproteins of very low and density + lipoproteins of low density; 2) lipoproteins of very low density + lipoproteins of low density; 3) only in lipoproteins of very low density. The frst one is specifc to piscivorous (carnivorous) while living in ocean. The second one is implemented by herbivorous while they didn\'t begin to synthesize insulin and hepatocytes not yet transform all endogenous fatty into oleic fatty . Insulin initiated: a) transferring of oleic fatty acids to lipoproteins of very low density without forming oleic lipoproteins of low density; b) highly effective oleic metabolism of fatty acids in vivo: c) becoming of biological function of locomotion. The aphysiological induction by substrate, surplus of fatty acids in food initiate negative alterations in composition of lipoproteins in opposite direction than in case of phylogenesis. When homo sapiens, herbivorous in phylogenesis, begins to misuse carnivorous (meat) food then instead of normolipoproteinemia in blood plasma under electrophoresis of lipoproteins one can initially detect transitory hyperlipoproteinemia type IV and then prolonged hyperlipoproteinemia type IIb. If patient factually passes on to carnivorous diet then hyperlipoproteinemia type V is developing. If content of exogenous fatty in food surpasses physiological capacities of its transferring in oleic triglycerides as palmitoyl-oleyl-palmitate glycerol, triglycerides as oleyl-palmitoyl-palmitate glycerol begin to form and epigenetically aphysiological non-ligand lipoproteins of very low density → lipoproteins of low density are formed. Their circulation in blood is a cause of hypertriglyceridemia, higher level of cholesterol-lipoproteins of low density, compensatory increasing of apoC-III. Then occurs induced by substrate formation of hyperlipoproteinemia initially of type IV, then of type IIb and fnally of type V. The pathogenesis of atherosclerosis and atheromotosis is activated when homo sapiens, herbivorous in phylogenesis, begin to misuse carnivorous food affecting biological functions of trophology, reaction of exotrophy (external nutrition), function of homeostasis, endoecology and function of adaptation. The formation of metabolism if fatty acids instead of oleic one is a cause of chronic defciency of and ATP synthesis in vivo. Insulin activates absorption of glucose by cells with purpose to use it for synthesis of oleic fatty acids. In the frst place, insulin regulates in vivo metabolism of fatty acids and only in second place metabolism of glucose.

Keyword: energy

Kinetics of coffee industrial residue pyrolysis using distributed activation model and components separation of bio-oil by sequencing temperature-raising pyrolysis.

This study was carried out to investigate the kinetics of coffee industrial residue (CIR) pyrolysis, the effect of pyrolysis factors on yield of bio-oil component and components separation of bio-oil. The kinetics of CIR pyrolysis was analyzed using distributed activation model (DAEM), based on the experiments in thermogravimetric analyzer (TGA), and it indicated that the average of activation (E) is 187.86kJ·mol. The bio-oils were prepared from CIR pyrolysis in vacuum tube furnace, and its components were determined by gas chromatography/mass spectrometry (GC-MS). Among pyrolysis factors, pyrolysis temperature is the most influential factor on components yield of bio-oil, directly concerned with the volatilization and yield of components (, linoleic , oleic , octadecanoic and caffeine). Furthermore, a new method (sequencing temperature-raising pyrolysis) was put forward and applied to the components separation of bio-oil. Based on experiments, a solution of components separation of bio-oil was come out.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: energy

Preparation and characterization of 100% bio-based polylactic / microcapsules for thermal storage.

Phase change materials (PCM) have gained extensive attention in thermal storage applications. In this work, microencapsulation of vegetable-derived (PA) in bio-based polylactic (PLA) shell by solvent evaporation and oil-in-water emulsification was investigated. Fourier transform infrared spectroscopy and scanning electron microscopy were conducted to confirm the successful encapsulation of PA in PLA shells. Differential scanning calorimetry was performed to evaluate the thermal properties, thermal reliability, and core content of the fabricated PCM microcapsules (microPCM). Through a series of parametric studies, the effects of PCM and solvent content, oil phase-to-aqueous phase ratio, as well as surfactant type and content on the morphology, particle size, and thermal properties of the PCM microcapsules were investigated. Experimental results showed that PVA was a superior emulsifier to SDS in the emulsion systems being studied. There also existed an optimal PVA concentration to reduce the average size of microPCM. When the PVA concentration was above this optimal level, the emulsifier molecules tend to form micelles among themselves. This led to the adhesion of tiny microspheres on the surface of microPCM as well as larger microPCM. In short, this work has demonstrated the possibility of using the solvent evaporation method to fabricate 100% bio-based PCM-polymer microcapsules for thermal storage applications.

Keyword: energy

Metabolic Profiles Reveal Changes in Wild and Cultivated Soybean Seedling Leaves under Salt Stress.

Clarification of the metabolic mechanisms underlying salt stress responses in plants will allow further optimization of crop breeding and cultivation to obtain high yields in saline-alkali land. Here, we characterized 68 differential metabolites of cultivated soybean (Glycine max) and wild soybean (Glycine soja) under neutral-salt and alkali-salt stresses using gas chromatography-mass spectrometry (GC-MS)-based metabolomics, to reveal the physiological and molecular differences in salt tolerance. According to comparisons of growth parameters under the two kinds of salt stresses, the level of inhibition in wild soybean was lower than in cultivated soybean, especially under alkali-salt stress. Moreover, wild soybean contained significantly higher amounts of phenylalanine, asparagine, citraconic , citramalic , citric and α-ketoglutaric under neutral-salt stress, and higher amounts of , lignoceric , glucose, citric and α-ketoglutaric under alkali-salt stress, than cultivated soybean. Further investigations demonstrated that the ability of wild soybean to salt tolerance was mainly based on the synthesis of organic and amino acids, and the more active tricarboxylic cycle under neutral-salt stress. In addition, the metabolite profiling analysis suggested that the generation from β-oxidation, glycolysis and the citric cycle plays important roles under alkali-salt stress. Our results extend the understanding of mechanisms involved in wild soybean salt tolerance and provide an important reference for increasing yields and developing salt-tolerant soybean cultivars.

Keyword: energy

Dietary sn-2 triacylglycerols reduced faecal lipids, calcium contents and altered lipid metabolism in Sprague-Dawley rats.

In this study, the impact of dietary sn-2 triacylglycerol (sn-2 PTAG) on faecal lipids, calcium excretion and lipid metabolic alternation was investigated in Sprague-Dawley (SD) rats fed with high-fat diet containing either palm olein (PO, sn-2 (PA) of 14.8%), sn-2 PTAG50 (sn-2\u2009PA of 56.4%) or sn-2 PTAG70 (sn-2\u2009PA of 72.4%), respectively. After 4-week feeding period, SD rats fed with sn-2 PTAGs showed reduced faecal soap fatty acids, neutral lipid and calcium excretion compared to those of PO-fed rats, whereas a significant difference was only observed for the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Moreover, dietary sn-2 PTAG70 also showed a significant effect on decreasing serum triacylglycerol (TAG) level, reducing perirenal adipocyte size and regulating lipid metabolism in small intestine and perirenal adipose tissue of SD rats. Significantly increased mRNA levels of genes involved in intestinal lipid anabolism as well as lipid catabolism were both observed in the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Meanwhile, dietary sn-2 PTAG70 also significantly up-regulated lipolysis, mitochondrial fatty oxidation and thermogenesis-related gene and protein levels in perirenal adipose tissue, which might be correlated with the reduced perirenal adipocyte size. Taken together, our findings indicated that sn-2 PTAG70 may have some beneficial effects on intestinal lipid utilisation and lipid metabolic activity for energy supply in visceral adipose tissue.

Keyword: energy

Fatty composition of free-living and parasitic stages of the bovine lungworm Dictyocaulus viviparus.

The development of parasitic nematodes proceeds via multiple stages, often implicating the necessity to adapt to different environments. Especially the transition from free-living to parasitic stages is accompanied by a significant change in the environmental conditions. To shed light on possible adaptations to these transitions, the fatty composition of different developmental stages of the bovine lungworm Dictyocaulus viviparus was investigated. Fatty acids of D. viviparus eggs, the free-living first, second and third larval stage (L1-L3) as well as the parasitic preadult stage and adult male and female worms residing in the lungs of infected hosts were quantified by gas chromatography after transesterification to their fatty methyl esters. The fatty content and diversity were higher in parasitic stages compared to those of free-living larvae. The most prevalent fatty acids in both parasitic and free-living stages were stearic (C18:0), (C16:0), palmitoleic (C16:1) and caprylic (C8:0). A variety of (poly-)unsaturated FAs was found in the parasitic stages and in the eggs, which was similar to the variety of FAs found in bovine surfactant. This finding indicates that parasitic stages of D. viviparus take up FAs from their environment. While eggs contained the highest concentration of fatty acids, a decrease was observed from eggs to L1 and further from L2 to L3. The lowest concentration was found in 38-days-old L3, which suggests that FAs serve as an reserve for the free-living, non-feeding larval stages. The free-living larvae contained mainly saturated fatty acids and only traces of unsaturated fatty acids, which is in contrast to the phospholipid saturation hypothesis of cold tolerance. Instead, a trade-off between desiccation stress and temperature adaptation may favour a higher amount of saturated FAs in the free-living larval stages. Further studies explicitly examining the FA composition of the different classes of lipids are necessary to better describe the adaptative responses of the FA metabolism to different environmental conditions.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: energy

Metabolic dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive oxygen species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal lipid loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different metabolic conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: energy

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in metabolism. However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: energy

Optimization of enzymatic hydrolysis of cellulosic fraction obtained from stranded driftwood feedstocks for lipid production by .

Stranded driftwood feedstocks may represent, after pretreatment with steam explosion and enzymatic hydrolysis, a cheap C-source for producing biochemicals and biofuels using oleaginous yeasts. The hydrolysis was optimized using a response surface methodology (RSM). The solid loading (SL) and the dosage of enzyme cocktail (ED) were variated following a central composite design (CCD) aimed at optimizing the conversion of carbohydrates into lipids (Y) by the yeast DBVPG 5870. A second-order polynomial equation was computed for describing the effect of ED and SL on Y. The best combination (ED\u202f=\u202f3.10%; SL\u202f=\u202f22.07%) for releasing the optimal concentration of carbohydrates which gave the highest predicted Y (27.32%) was then validated by a new hydrolysis. The resulting value of Y (25.26%) was close to the theoretical maximum value. Interestingly, fatty profile achieved under the optimized conditions was similar to that reported for palm oil.

Keyword: energy

Functional lipidomics: impairs hepatocellular carcinoma development by modulating membrane fluidity and glucose metabolism.

Lipids are essential cellular components and sources of living organisms, and altered lipid composition is increasingly recognized as a signature of cancer. We performed lipidomic analysis in a series of hepatocellular carcinoma (HCC) cells and identified over 1,700 intact lipids originating from three major lipid categories. Comparative lipidomic screening revealed that 93 significantly changed lipids and decreased acyl (C16:0)-containing glycerophospholipids were positively associated with metastatic abilities of HCC cells. Furthermore, both in vitro and in vivo experiments demonstrated that C16:0 incubation specifically reduced malignant cell proliferation, impaired cell invasiveness, and suppressed tumor growth in mouse xenograft models. Biochemical experiments demonstrated that C16:0 treatment decreased cell membrane fluidity and limited glucose metabolism. A phosphoproteomics approach further revealed such C16:0 incubation attenuated phosphorylation levels of mammalian target of rapamycin (mTOR) and signal transducer and activator of transcription 3 (STAT3) pathway proteins. Multiple reaction monitoring analysis of 443 lipid molecules showed 8 reduced C16:0-containing lipids out of total 10 altered lipids when cancer tissues were compared with adjacent nontumor tissues in a cohort of clinical HCC specimens (P < 0.05).These data collectively demonstrate the biomedical potential of using altered lipid metabolism as a diagnostic marker for cancerous cells and open an opportunity for treating aggressive HCCs by targeting altered C16:0 metabolism. (Hepatology 2017;66:432-448).© 2017 by the American Association for the Study of Liver Diseases.

Keyword: energy

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: energy

Good Fats versus Bad Fats: A Comparison of Fatty Acids in the Promotion of Insulin Resistance, Inflammation, and Obesity.

Recently, debate has erupted in both the scientific community and throughout the lay public around whether a low-fat or low-carbohydrate diet is better for weight loss. In other words, is it better to cut fat or cut carbohydrate for weight loss. However, going beyond this debate (fat versus carbohydrate), are questions around whether certain fatty acids are worse for promoting insulin resistance, inflammation, and obesity. The overall evidence in the literature suggests that medium-chain saturated fats (such as lauric , found in coconut oil) and monounsaturated fat (oleic , found in olive oil) are less likely to promote insulin resistance, inflammation, and fat storage compared to long-chain saturated fatty acids (such as stearic found in large quantities in butter, but particularly found in palm oil) especially when consumed on top of a diet moderate in refined carbohydrates. Compared to long-chain saturated fats, lauric and oleic have an increased fatty oxidation rate, are more likely to be burned for and less likely to be stored in adipose tissue, and thus promote increased expenditure. Omega-6 polyunsaturated fatty acids (PUFAs), such as linoleic , as found in vegetable oils may contribute to obesity, whereas omega-3 PUFA may be protective. Importantly, both olive oil as part of a Mediterranean diet, and omega-3 from fish and fish oil have been proven to reduce risk of cardiovascular (CV) events.

Keyword: energy

Chemical and Nutritional Composition of Terminalia ferdinandiana (Kakadu Plum) Kernels: A Novel Nutrition Source.

(Kakadu plum) is a native Australian fruit. Industrial processing of fruits into puree generates seeds as a by-product, which are generally discarded. The aim of our present study was to process the seed to separate the kernel and determine its nutritional composition. The proximate, mineral and fatty compositions were analysed in this study. Kernels are composed of 35% fat, while proteins account for 32% dry weight (DW). The content and fiber were 2065 KJ/100 g and 21.2% DW, respectively. Furthermore, the study showed that kernels were a very rich source of minerals and trace elements, such as potassium (6693 mg/kg), calcium (5385 mg/kg), iron (61 mg/kg) and zinc (60 mg/kg) DW, and had low levels of heavy metals. The fatty composition of the kernels consisted of omega-6 fatty , linoleic (50.2%), monounsaturated oleic (29.3%) and two saturated fatty acids namely (12.0%) and stearic (7.2%). The results indicate that kernels have the potential to be utilized as a novel protein source for dietary purposes and non-conventional supply of linoleic, and oleic acids.

Keyword: energy

Fabrication and Performance of Composite Microencapsulated Phase Change Materials with Ethyl Ester as Core.

Microencapsulation of phase change materials (PCMs) could prevent the leakage of PCMs during solid⁻liquid phase change process. However, their applications are mainly limited by the compactness and thermal stability of the traditional polyurea shell microcapsules. To increase the thermal compactness and thermal stability of PCM microcapsules, tetraethylorthosilicate (TEOS) was employed to form polymer/SiO₂ composite shells to enhance the mechanical performance of polyurea and polyurethane microcapsule via interfacial polymerization and in situ polymerization. The morphology and chemical components of the microcapsules were characterized by field-emission scanning electron microscope (FE-SEM) and Fourier transform infrared (FT-IR) spectroscopy, respectively. The thermal properties of the microcapsules were investigated by differential scanning calorimetry (DSC) and thermal gravity analysis (TGA). The results showed the smoothness and compactness of both polyurea⁻SiO₂ and polyurethane⁻SiO₂ microcapsules enhanced slightly, when compared with that without TEOS addition. Moreover, the SiO₂ composite shell had good effect on thermal compactness, as the weight loss rate of polyurea⁻SiO₂ microcapsules and polyurethane⁻SiO₂ microcapsules decreased 3.5% and 4.1%, respectively.

Keyword: energy

Green preparation of seaweed-based silver nano-liquid for cotton pathogenic fungi management.

Silver nanoparticles (Ag NPs) were synthesised using the crude ethyl acetate extracts of and evaluated their bioefficacy against two crop-damaging pathogens. The sets of lattice planes in the XRD spectrum for the Ag NPs were indexed to the 111, 200, 220 and 311 orientations and support the crystalline nature of the Ag NPs. The 3414 and 2968\u2005cm peaks were observed in crude algal thallus extract and they were characteristic of terpenoids. Further, a peak at 1389\u2005cm was observed as fatty acids. The marine macroalgae terpenoids and acted as reducing agent and stabiliser, respectively. The size (3 and 50\u2005nm) and shape (spherical) of Ag NPs were recorded. The -dispersive X-ray spectroscopy analysis exemplified the presence of silver in its elemental nature. Moreover, Ag NPs were effective against two cotton phytopathogens namely f.sp. (FOV) and pv. malvacearum (XAM). The minimum inhibitory concentration was found to be 80.0 and 43.33\u2005μg ml against FOV and XAM, respectively. Results confirmed the anti-microbial activity of green nanoparticles against select pathogens and suggest their possible usage in developing antifungal agents for controlling destructive pathogens in a cotton agroecosystem.

Keyword: energy

Dehydration Causes Increased Reliance on Protein Oxidation in Mice: A Test of the Protein-for-Water Hypothesis in a Mammal.

During fasting, animals rely on a mixture of fats, carbohydrates, and proteins that are derived solely from endogenous sources. The relative contributions of these metabolic fuels chiefly depend on the duration of the fast, but other factors including previous diet, environmental temperature, and activity level can modulate the fuel mixture. It has long been held that endogenous proteins are spared from catabolism until the final stages of prolonged fasting and contribute a significant proportion of once the other metabolic fuels have been depleted. However, evidence is mounting that protein is catabolized supplemental to fat metabolism under some circumstances. This has been shown in migratory birds that exhibit dramatic reductions in lean mass during flights. One hypothesis to explain this seemingly maladaptive metabolic strategy is that the catabolism and oxidation of protein in situ yields five times more metabolic water than that generated through fat oxidation alone. Recent support for this hypothesis is that birds benefit from such a strategy due to their uricotelic nature. However, it remains unclear whether ureotelic mammals would also employ this strategy. Here we test the protein-for-water hypothesis in resting laboratory mice subjected to water deprivation during fasting while we tracked rates of protein and lipid catabolism using endogenously incorporated C-leucine and C-. We found no differences in instantaneous leucine oxidation; however, cumulative differences in instantaneous leucine oxidation ultimately resulted in a higher total leucine oxidation after 72 h of fasting in water-deprived animals. We also found that lipid oxidation was 8% higher in the hydrated mice, but the difference was not significant presumably because of a concomitant reduction in metabolic rates of the water-deprived mice. Our results indicate that mammals do increase rates of protein catabolism during dehydration but to a lesser degree than birds. The ability of mammals to produce highly concentrated urine and their lower inherent rates of protein turnover apparently preclude mammals from taking full advantage of the protein-for-water strategy during fasting under dehydrating conditions.

Keyword: energy

Fatty profiles of the European migratory common noctule bat (Nyctalus noctula).

In animals, fatty acids (FA) are essential as structural components in membranes and for storage in adipocytes. Here, we studied the relative proportions of FA in a mammal with extreme changes in metabolic rates. Common noctule bats (Nyctalus noctula) switch from energetically demanding long-distance migration at high metabolic rates to regular torpor with extremely low metabolic rates. We found that composition of FA categories differed between adipose tissue types (white adipose tissue (WAT) vs brown adipose tissue (BAT)) and muscle tissue types (skeletal vs heart), but not between sexes. We found oleic to be the most abundant FA in all studied tissues. Concentrations of polyunsaturated FA (PUFA) were not always higher in muscular tissue compared with adipocyte tissue, even though high concentrations of PUFA are considered beneficial for low body temperatures in torpor. In all tissues, we observed a high content in monounsaturated fatty acids (MUFA), possibly to compensate for a low PUFA content in the diet. Ratios of ω6/ω3 were lower in the heart than in skeletal muscles of common noctules. Three FA (, oleic, and linoleic ) accounted for about 70% of the FA in adipose tissue, which is similar to proportions observed in migrating birds, yet migrating birds generally have a higher PUFA content in muscle and adipose tissues than bats. Bats seem to contrast with other mammals in having a high MUFA content in all tissues. We conclude that FA profiles of bats differ largely from those of most cursorial mammals and instead are-with the exception of MUFA-similar to those of migrating birds.

Keyword: energy

Altering the ratio of dietary C16:0 and cis-9 C18:1 interacts with production level in dairy cows: Effects on production responses and partitioning.

The objective of our study was to evaluate the effects of altering the dietary ratio of (C16:0) and oleic (cis-9 C18:1) acids on nutrient digestibility, partitioning, and production responses of lactating dairy cows. Cows were blocked by milk yield and assigned to 3 groups (12 cows per group) in a main plot: low (45.2 ± 1.7 kg/d), medium (53.0 ± 1.6 kg/d), and high (60.0 ± 1.9 kg/d). Within each production group, a truncated Latin square arrangement of fatty (FA) treatments was used in 2 consecutive 35-d periods. The FA treatments supplemented at 1.5% of diet dry matter were (1) 80:10 (80% C16:0 + 10% cis-9 C18:1), (2) 73:17 (73% C16:0 + 17% cis-9 C18:1), (3) 66:24 (66% C16:0 + 24% cis-9 C18:1), and (4) 60:30 (60% C16:0 + 30% cis-9 C18:1). Treatment × production group interactions were observed for yields of milk, fat-corrected milk, -corrected milk, milk fat, milk protein, and milk lactose and partitioned to milk. Increasing cis-9 C18:1 in FA treatments reduced fat-corrected milk, -corrected milk, and milk output in low-producing cows but increased these in high-producing cows. Increasing cis-9 C18:1 in FA treatments did not affect milk yield, milk protein yield, and milk lactose yield in low- and medium-producing cows but increased these in high-producing cows. Regardless of production level, there was no effect of treatments on dry matter intake; however, increasing cis-9 C18:1 in FA treatments increased body weight change and body condition score change. Increasing cis-9 C18:1 in FA treatments increased total FA digestibility due to a linear increase in 16- and 18-carbon FA digestibilities. Interactions between FA treatments and production level were observed for the yield of milk fat and milk FA sources. In low-producing cows, increasing cis-9 C18:1 in FA treatments decreased milk fat yield due to a decrease in de novo and mixed milk FA without changes in preformed milk FA. In contrast, in high-producing cows, increasing cis-9 C18:1 in FA treatments increased milk fat yield due to an increase in de novo and preformed milk FA. Our results indicate that high-producing dairy cows (averaging 60 kg/d) responded better to a fat supplement containing more cis-9 C18:1, whereas low-producing cows (averaging 45 kg/d) responded better to a supplement containing more C16:0.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Linoleic rescues microglia inflammation triggered by saturated fatty .

Elevated saturated free fatty levels during over-nutrition lead to hypothalamic inflammation, which perturbs homeostasis. Whether brain-derived metabolites are coupled to the development of obesity pathogenesis during the early over-nutrition period has not been thoroughly investigated. In this study, we found increased linoleic , an unsaturated fatty , in both the whole brain and hypothalamus of mice fed a high-fat diet for 4 weeks. Furthermore, we observed that linoleic effectively reversed the inflammatory responses induced by treatment in microglial cells. Collectively, this study suggests the reversible function of linoleic on brain inflammation in association with microglial activation during short-term exposure to a high-fat diet.Copyright © 2019. Published by Elsevier Inc.

Keyword: energy

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance.

Patients with type 2 diabetes (T2D) and/or insulin resistance (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the insulin resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased insulin-stimulated glucose and FA uptake, as well as loss of endosomal acidification and insulin signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to obesity, diabetes and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Adaptations of hepatic lipid metabolism and mitochondria in dairy cows with mild fatty liver.

The inevitable deficiency in nutrients and at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. Fatty liver is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and mitochondria in dairy cows with mild fatty liver, liver and blood samples were collected from healthy cows (n = 15) and cows with mild fatty liver (n = 15). To determine the effects of acids (PA), one of the major component of fatty acids, on lipid metabolism and mitochondria in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM). Dairy cows with mild fatty liver displayed hepatic lipid accumulation. The protein levels of sterol regulatory element-binding protein 1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPARα) and mRNA levels of acetyl CoA carboxylase 1 (ACC1), fatty synthase (FAS), acyl-CoA oxidase (ACO), and carnitine palmitoyltransferase 1A (CPT1A) were significantly higher in dairy cows with mild fatty liver than in control cows. The hepatic mitochondrial DNA content, mRNA levels of oxidative phosphorylation complexes I to V (CO 1-V), protein levels of cytochrome c oxidase subunit IV (COX IV), voltage dependent anion channel 1 (VDAC1), peroxisome proliferator activated receptor-γ coactivator-1α (PGC-1α) and nuclear respiratory factor 1 (NRF1), and adenosine triphosphate (ATP) content were all markedly increased in the liver of dairy cows with mild fatty liver compared with healthy cows. The PA treatment significantly increased lipid accumulation; protein levels of SREBP-1c and PPARα; and mRNA levels of ACC1, FAS, ACO, and CPT1A in calf hepatocytes. Moreover, the mitochondrial DNA content, mRNA levels of CO 1-V, protein levels of COX IV, VDAC1, PGC-1α, NRF1, mitochondrial transcription factor A, and ATP content were significantly increased in PA-treated hepatocytes compared with control hepatocytes. The protein level of mitofusin-2 was significantly decreased in PA-treated groups. In conclusion, lipid synthesis and oxidation, number of mitochondria, and ATP production were increased in the liver of dairy cows with mild fatty liver and PA-treated calf hepatocytes. These changes in hepatic mitochondria and lipid metabolism may be the adaptive mechanism of dairy cows with mild fatty liver.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Exploring the potency of integrating semi-batch operation into lipid yield performance of Chlamydomonas sp. Tai-03.

Third generation biofuels, also known as microalgal biofuels, are promising alternatives to fossil fuels. One attractive option is microalgal biodiesel as a replacement for diesel fuel. Chlamydomonas sp. Tai-03 was previously optimized for maximal lipid production for biodiesel generation, achieving biomass growth and productivity of 3.48\u202f±\u202f0.04\u202fg/L and 0.43\u202f±\u202f0.01\u202fg/L/d, with lipid content and productivity of 28.6\u202f±\u202f1.41% and 124.1\u202f±\u202f7.57\u202fmg/L/d. In this study, further optimization using 5% CO concentration and semi-batch operation with 25% medium replacement ratio, enhanced the biomass growth and productivity to 4.15\u202f±\u202f0.12\u202fg/L and 1.23\u202f±\u202f0.02\u202fg/L/d, with lipid content and productivity of 19.4\u202f±\u202f2.0% and 239.6\u202f±\u202f24.8\u202fmg/L/d. The major fatty methyl esters (FAMEs) were (C16:0), oleic (C18:1), and linoleic (C18:2). These short-chain FAMEs combined with high growth make Chlamydomonas sp. Tai-03 a suitable candidate for biodiesel synthesis.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

Short communication: Comparison of a -enriched triglyceride supplement and calcium salts of palm fatty acids supplement on production responses of dairy cows.

The objective of our study was to evaluate the effects of feeding a -enriched triglyceride supplement or a calcium salts of palm fatty (FA) supplement on nutrient digestibility and production responses of mid-lactation dairy cows. Fifteen Holstein cows (139 ± 39 d in milk) were randomly assigned to treatment sequence in a 3 × 3 Latin square design. Treatments were a control diet (CON; no fat supplement) and 1.5% of FA added either as a -enriched triglyceride supplement (PA-TG) or as calcium salts of palm FA supplement (Ca-FA). Fat-supplemented treatments did not affect dry matter intake (DMI) compared with CON, but Ca-FA reduced DMI compared with PA-TG. Compared with CON, fat-supplemented treatments increased 18-carbon FA digestibility by 2.0 percentage units but did not affect digestibility of total FA or 16-carbon FA. Compared with Ca-FA, PA-TG reduced total FA digestibility by 8.7 percentage units due to a decrease in 16-carbon FA digestibility (21.7 percentage units). Both fat supplements increased neutral detergent fiber (NDF) digestibility compared with CON (3.90 percentage units), and PA-TG tended to increase NDF digestibility by 1.60 percentage units compared with Ca-FA. Compared with CON, fat-supplemented treatments increased milk yield (1.05 kg/d), 3.5% fat-corrected milk yield (2.20 kg/d), and -corrected milk yield (1.80 kg/d). Also, PA-TG increased milk fat yield (50 g/d) and milk output (1.0 Mcal/d) and tended to increase milk fat content (0.07 percentage units) and -corrected milk yield (1.0 kg/d) compared with Ca-FA. Fat-supplemented treatments reduced the yield of de novo milk FA (23 g/d) and increased the yields of mixed (43 g/d) and preformed (52 g/d) milk FA compared with CON. The PA-TG treatment increased the yield of 16-carbon (66 g/d) milk FA compared with Ca-FA, whereas Ca-FA increased the yield of preformed (60 g/d) milk FA. Fat-supplemented treatments increased intake of net for lactation by 1.80 Mcal/d, milk output by 1.30 Mcal/d, and in body reserves by 0.30 Mcal/d compared with CON. The Ca-FA treatment increased allocated to body reserves (0.60 Mcal/d), partitioning toward body reserves (1.20 percentage units), and body condition score change (0.06 units), and tended to increase body weight change (0.16 kg/d) and body condition score (0.08 units) compared with PA-TG. In conclusion, feeding a -enriched triglyceride supplement increased milk output due to increased yields of milk and milk fat, whereas feeding a calcium salts of palm FA supplement increased FA digestibility and partitioned to body reserves.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: energy

Resveratrol Ameliorates Lipid Droplet Accumulation in Liver Through a SIRT1/ ATF6-Dependent Mechanism.

Lipid droplets (LDs) are dynamic organelles that store neutral lipids during times of excess, and an increased accumulation of LDs in the liver is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high-fat diet (HFD) and were used to induce hepatic steatosis in mouse liver and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in liver and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: energy

Sodium-carboxylate contact ion pair formation induces stabilization of monolayers at high pH.

Sea spray aerosols (SSA) are known to have an organic coating that is mainly composed of fatty acids. In this study, the effect of pH and salt on the stability and organization of a (PA) monolayer is investigated by surface vibrational spectroscopy and molecular dynamics simulations. Results indicate that alkyl chain packing becomes more disordered as the carboxylic headgroup becomes deprotonated. This is associated with packing mismatch of charged and neutral species as charged headgroups penetrate deeper into the solution phase. At pH 10.7, when the monolayer is ∼99% deprotonated, palmitate (PA) molecules desorb and solubilize into the bulk solution where there is spectroscopic evidence for aggregate formation. Yet, addition of 100 mM NaCl to the bulk solution is found to drive PA molecules to the aqueous surface. Free calculations show that PA molecules become stabilized within the interface with increasing NaCl concentration. Formation of contact -COO:Na pairs alters the hydration state of PA headgroups, thus increasing the surface propensity. As salts are highly concentrated in SSA, these results suggest that deprotonated fatty acids may be found at the air-aqueous interface of aerosol particles due to sea salt\'s role in surface stabilization.

Keyword: energy

The selective production of jet fuel range alkanes via the catalytic upgrading of over Co/HMCM-49 catalysts.

The complete deoxygenation of followed by simultaneous hydro-isomerization and appropriate hydrocracking were achieved over Co/HMCM-49 bi-functional catalysts with the special structure of 12 MR cups and a low Si/Al ratio. This offered new insights into the design of bi-functional catalysts for carbon rearrangements during the upgrading of to jet fuel range alkanes.

Keyword: energy

Own-Synthetize Nanoparticles to Develop Nano-Enhanced Phase Change Materials (NEPCM) to Improve the Efficiency in Buildings.

The use of adequate thermal storage (TES) systems is an opportunity to increase efficiency in the building sector, and so decrease both commercial and residential consumptions. Nano-enhanced phase change materials (NEPCM) have attracted attention to address one of the crucial barriers (i.e. low thermal conductivity) to the adoption of phase change materials (PCM) in this sector. In the present study two PCM based on fatty acids, capric and , were nano-enhanced with low contents (1.0 wt.%, 1.5 wt.% and 3.0 wt.%) of copper (II) oxide (CuO) nanoparticles. Copper (II) oxide (CuO) was synthesized via coprecipitation method obtaining 60⁻120 nm diameter sized nanoparticles. Thermal stability and high thermal conductivity were observed for the nano-enhanced phase change materials (NEPCM) obtained. Experimental results revealed remarkable increments in NEPCM thermal conductivity, for instance thermal conductivity was increased up to 60% with the addition of 3 wt.% CuO nanoparticles. Moreover, CuO nanoparticles sedimentation velocity decreases when increasing its content.

Keyword: energy

stimulates metabolism and inhibits insulin/PI3K/AKT signaling in differentiated human neuroblastoma cells: The role of mTOR activation and mitochondrial ROS production.

The high consumption of saturated lipids has been largely associated with the increasing prevalence of metabolic diseases. In particular, saturated fatty acids such as (PA) have been implicated in the development of insulin resistance in peripheral tissues. However, how neurons develop insulin resistance in response to lipid overload is not fully understood. Here, we used cultured rat cortical neurons and differentiated human neuroblastoma cells to demonstrate that PA blocks insulin-induced metabolic activation, inhibits the activation of the insulin/PI3K/Akt pathway and activates mTOR kinase downstream of Akt. Despite the fact that fatty acids are not normally used as a significant source of fuel by neural cells, we also found that short-term neuronal exposure to PA reduces the NAD/NADH ratio, indicating that PA modifies the neuronal balance. Finally, inhibiting mitochondrial ROS production with mitoTEMPO prevented the deleterious effect of PA on insulin signaling. This work provides novel evidence of the mechanisms behind saturated fatty -induced insulin resistance and its metabolic consequences on neuronal cells.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Seed traits, fatty profile and genetic diversity assessment in Pongamia pinnata (L.) Pierre germplasm.

Phenotypic variation of important seed traits like seed length, seed breadth, seed thickness, 100 seed weight and seed oil content were recorded in a total of 157 collected accessions of Pongamia. Out of these, fatty profiles of 38 accessions selected based on their high and low oil content was analyzed. Fatty profile revealed high variability in stearic, oleic and linoleic which varied from 0.42 to 10.61\xa0%, 34.34 to 74.58\xa0%, and 7.00 to 31.28\xa0% respectively. Variations in and linolenic were small. Iodine value, saponification number and cetane number (CN) of fatty methyl esters (FAME) of seed oil ranges from 186.99 to 201.25, 81.13 to 108.19 and 46.16 to 56.47 respectively. Fatty compositions, degree of unsaturation and CN are the important parameters, which are used to determine quality of FAME were used as biodiesel. Some of the Pongamia accessions identified were higher in oil content while some accessions showed higher degree of unsaturation and a few of them had CN values higher than 55. Genetic diversity analysis with six TE-AFLP primers generated a total of 334 bands out of which 174 (52.10\xa0%) were polymorphic. The genetic similarity ranged from 0.11 to 0.47. These findings clearly showed high level of genetic diversity and all economically desirable traits were not present in a single genotype of Pongamia. All these traits could be selected from these CPTs and transfer to a single elite variety through selection and breeding programme and could be utilized for large scale multiplication and plantation to produce high quantity and quality biodiesel in future.

Keyword: energy

Wax Ester Fermentation and Its Application for Biofuel Production.

In Euglena cells under anaerobic conditions, paramylon, the storage polysaccharide, is promptly degraded and converted to wax esters. The wax esters synthesized are composed of saturated fatty acids and alcohols with chain lengths of 10-18, and the major constituents are myristic and myristyl alcohol. Since the anaerobic cells gain ATP through the conversion of paramylon to wax esters, the phenomenon is named "wax ester fermentation". The wax ester fermentation is quite unique in that the end products, i.e. wax esters, have relatively high molecular weights, are insoluble in water, and accumulate in the cells, in contrast to the common fermentation end products such as lactic and ethanol.A unique metabolic pathway involved in the wax ester fermentation is the mitochondrial fatty synthetic system. In this system, fatty are synthesized by the reversal of β-oxidation with an exception that trans-2-enoyl-CoA reductase functions instead of acyl-CoA dehydrogenase. Therefore, acetyl-CoA is directly used as a C donor in this fatty synthesis, and the conversion of acetyl-CoA to malonyl-CoA, which requires ATP, is not necessary. Consequently, the mitochondrial fatty synthetic system makes possible the net gain of ATP through the synthesis of wax esters from paramylon. In addition, acetyl-CoA is provided in the anaerobic cells from pyruvate by the action of a unique enzyme, oxygen sensitive pyruvate:NADP oxidoreductase, instead of the common pyruvate dehydrogenase multienzyme complex.Wax esters produced by anaerobic Euglena are promising biofuels because myristic (C) in contrast to other algal produced fatty acids, such as (C) and stearic (C), has a low freezing point making it suitable as a drop-in jet fuel. To improve wax ester production, the molecular mechanisms by which wax ester fermentation is regulated in response to aerobic and anaerobic conditions have been gradually elucidated by identifying individual genes related to the wax ester fermentation metabolic pathway and by comprehensive gene/protein expression analysis. In addition, expression of the cyanobacterial Calvin cycle fructose-1,6-bisphosphatase/sedohepturose-1,7-bisphosphatase, in Euglena provided photosynthesis resulting in increased paramylon accumulation enhancing wax ester production. This chapter will discuss the biochemistry of the wax ester fermentation, recent advances in our understanding of the regulation of the wax ester fermentation and genetic engineering approaches to increase production of wax esters for biofuels.

Keyword: energy

Bi-allelic mutations of LONP1 encoding the mitochondrial LonP1 protease cause pyruvate dehydrogenase deficiency and profound neurodegeneration with progressive cerebellar atrophy.

LonP1 is crucial for maintaining mitochondrial proteostasis and mitigating cell stress. We identified a novel homozygous missense LONP1 variant, c.2282 C\xa0>\xa0T, (p.Pro761Leu), by whole-exome and Sanger sequencing in two siblings born to healthy consanguineous parents. Both siblings presented with stepwise regression during infancy, profound hypotonia and muscle weakness, severe intellectual disability and progressive cerebellar atrophy on brain imaging. Muscle biopsy revealed the absence of ragged-red fibers, however, scattered cytochrome c oxidase-negative staining and electron dense mitochondrial inclusions were observed. Primary cultured fibroblasts from the siblings showed normal levels of mtDNA and mitochondrial transcripts, and normal activities of oxidative phosphorylation complexes I through V. Interestingly, fibroblasts of both siblings showed glucose-repressed oxygen consumption compared to their mother, whereas galactose and utilization were similar. Notably, the siblings\' fibroblasts had reduced pyruvate dehydrogenase (PDH) activity and elevated intracellular lactate:pyruvate ratios, whereas plasma ratios were normal. We demonstrated that in the siblings\' fibroblasts, PDH dysfunction was caused by increased levels of the phosphorylated E1α subunit of PDH, which inhibits enzyme activity. Blocking E1α phosphorylation activated PDH and reduced intracellular lactate concentrations. In addition, overexpressing wild-type LonP1 in the siblings\' fibroblasts down-regulated phosphoE1α. Furthermore, in vitro studies demonstrated that purified LonP1-P761L failed to degrade phosphorylated E1α, in contrast to wild-type LonP1. We propose a novel mechanism whereby homozygous expression of the LonP1-P761L variant leads to PDH deficiency and metabolism dysfunction, which promotes severe neurologic impairment and neurodegeneration.

Keyword: energy

Flaxseed oil ameliorated high-fat-diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts.

α-Linolenic (ALA) is a plant-derived omega-3 unsaturated fatty that is rich in flaxseed oil (FO). The effect of FO on bone health is controversial. This study aims to evaluate the effect of FO on bone damage induced by a high-fat diet (HFD) and to explore the possible mechanism.Male Sprague-Dawley rats were fed a normal control diet (NC, 10% fat), FO diet (NY, 10% fat), HFD (60% fat), or HFD containing 10% FO (HY, 60% fat) for 22\u2009weeks. Micro CT and three-point bending tests were conducted to evaluate bone microstructure and biomechanics. Serum was collected for the detection of ALP, P1NP, and CTX-1. Rat primary osteoblasts (OBs) were treated with different concentrations of ALA with or without (PA) treatment. The ALP activity, osteogenic-related gene and protein expression were measured.Rats in the HFD group displayed decreased biomechanical properties, such as maximum load, maximum fracture load, ultimate tensile strength, stiffness, absorption, and elastic modulus, compared with the NC group ( \xa0<\u20090.05). However, HY attenuated the HFD-induced decreases in bone biomechanical properties, including maximum load, maximum fracture load, and ultimate tensile strength (\u2009<\u20090.05). Trabecular bone markers such as trabecular volume bone mineral density (Tb. vBMD trabecular bone volume/total volume (Tb. BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) were decreased, trabecular separation (Tb. Sp) and the structure model index (SMI) were increased in the HFD group compared with the NC group, and all parameters were remarkably improved in the HY group compared to the HFD group (p\u2009<\u20090.05). However, cortical bone markers such as cortical volume bone mineral density (Ct. vBMD), cortical bone volume/total volume (Ct. BV/TV) and cortical bone thickness (Ct. Th) were not significantly different among all groups. Moreover, the serum bone formation markers ALP and P1NP were higher and the bone resorption marker CTX-1 was lower in the HY group compared with levels in the HFD group. Compared with the NC group, the NY group had no difference in the above indicators. In rat primary OBs, PA treatment significantly decreased ALP activity and osteogenic gene and protein (β-catenin, RUNX2, and osterix) expression, and ALA dose-dependently restored the inhibition induced by PA.FO might be a potential therapeutic agent for HFD-induced bone loss, most likely by promoting osteogenesis.© The Author(s). 2019.

Keyword: energy

Role of the saturated fatty palmitate in the interconnected hypothalamic control of homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, obesity, and type 2 diabetes mellitus. Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: energy

Binding energies and the entry route of and palmitoylcarnitine into myoglobin.

The interaction of lipids (entry mechanism) with respect to both oxy- and deoxy-myoglobin was explored using unrestrained Molecular Dynamics simulations. The results indicated a spontaneous entry of both and palmitoylcarnitine molecules into the oxy-Mb structure at the main binding site, whereas in deoxy-Mb, both the lipid ligands move away from the protein surface. For the alternative binding locations, entry of the ligands was independent of the oxygenation state. Presented here are the tables with the myoglobin binding energies for and palmitoylcarnitine estimated using Alchemical Free Perturbation approach for the key structures obtained in unrestrained Molecular Dynamics simulations. These data are referenced in the original article "Exploring the entry route of and palmitoylcarnitine into myoglobin", reference number YABBI7787.

Keyword: energy

A promising form-stable phase change material prepared using cost effective pinecone biochar as the matrix of for thermal storage.

A promising new form-stable phase change material (PA/PB) was fabricated using pinecone biochar (PB) as the supporting material of (PA). The biochar of PB with large surface area was produced by forest residue of pinecone, and it was cheap, environment friendly and easy to prepare. The PB was firstly utilized as the supporter of PA and the characterizations of PA/PB were analyzed by the BET, SEM, XRD, DSC, TGA, FT-IR and thermal conductivity tester. The results demonstrated that the PA was physically absorbed by the PB and the crystal structure of the PA was not destroyed. The results of DSC showed that the fusing and crystallization points of the form-stable phase change material with the maximum content of PA (PA/PB-4) were 59.25\u2009°C and 59.13\u2009°C, and its fusing and freezing latent heat were 84.74\u2009kJ/kg and 83.81\u2009kJ/kg, respectively. The results of TGA suggested that the thermal stability of the PA/PB-4 composite was excellent, which could be used for the applications of thermal storage. Furthermore, the thermal conductivity of PA/PB-4 was 0.3926\u2009W/(m∙K), which was increased by 43.76% compared with that of the pure PA. Thus, the study results indicated that the PA/PB-4 had great potential for thermal storage applications.

Keyword: energy

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic metabolism-mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 diabetes.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Association of low oleic intake with diabetic retinopathy in type 2 diabetic patients: a case-control study.

The objective of this study was to describe the intake of macronutrient, especially fatty acids, and explore their possible effect on diabetic retinopathy (DR) in patients with type 2 diabetes mellitus.In this case-control study, we included a total of 146 patients with DR and 148 without DR. The intake of macronutrient was evaluated using a validated food frequency questionnaire. We used logistic regression adjusted for sex, age, diabetes duration, intake, educational level, physical activity, waist circumference, systolic blood pressure, high-density lipoprotein cholesterol and diabetes treatment, to estimate odds ratio (ORs) of DR.Patients with DR had significantly lower intake of fibre, monounsaturated fatty acids (MUFA), and and oleic . Inverse associations were observed between MUFA and oleic intake in DR. Subjects with intermediate and high MUFA intake were less likely to have DR than those with lower MUFA intake, with ORs of 0.46 (95\u2009%\u2009CI: 0.22-0.93) and 0.42 (95\u2009%\u2009CI: 0.18-0.97), respectively. Similarly, intermediate and high oleic intake were associated with reduced DR frequency compared with low oleic intake, with OR values of 0.48 (95\u2009%\u2009CI: 0.23-0.97) and 0.37 (95\u2009%\u2009CI: 0.16-0.85), respectively. These associations were stronger in patients with a longer diabetes duration.In type 2 diabetes mellitus, MUFA and oleic intake were inversely associated with DR.

Keyword: energy

Palmitate Induces an Anti-Inflammatory Response in Immortalized Microglial BV-2 and IMG Cell Lines that Decreases TNFα Levels in mHypoE-46 Hypothalamic Neurons in Co-Culture.

Elevated levels of saturated fatty acids (SFA) induce a state of neuroinflammation in the hypothalamus. It has been suggested that microglia sense palmitate, a prevalent circulating SFA, and act as mediators of this inflammatory process by communicating with neurons, particularly those involved in appetite regulation. In this study, we examined the inflammatory response to palmitate in immortalized microglial cell lines, BV-2 and IMG, and the subsequent effects on inflammatory gene expression in a model of NPY/AgRP neurons, mHypoE-46.The BV-2 cells were treated with 50 µM palmitate for 4 and 24 h, and the transcriptional regulation of markers for inflammation and cellular stress was assessed using an RT2 Profiler PCR Array. Select genes were verified with qRT-PCR. The BV-2 and IMG cells were then co-cultured using 1.0-µm cell culture inserts with an immortalized hypothalamic cell line, mHypoE-46, to investigate potential intercellular communication between microglia and neurons.We found that palmitate increased the mRNA levels of specific inflammatory genes, and a general anti-inflammatory profile was revealed in the microglia cells. The mRNA changes in TNFα at 4 and 24 h in BV-2 cells were abrogated with the toll-like receptor 4 (TLR4) inhibitor, TAK-242, indicating the involvement of TLR4. Co-culture of mHypoE-46 neurons with microglia pre-treated with palmitate resulted in repression of TNFα expression in the hypothalamic neurons. As palmitate significantly increased IL-13 expression in microglia, the effect of this cytokine was tested in mHypoE-46 neurons. The addition of IL-13 to neuronal cultures normalized the palmitate-mediated increase in IL-6 and AgRP expression, suggesting that microglia may protect surrounding neurons, at least in part, through the release of IL-13.These results suggest a potential anti-inflammatory role of microglia towards the palmitate-induced neuroinflammation, and potentially homeostasis, in hypothalamic neurons.© 2018 ©2018S. Karger AG, Basel.

Keyword: energy

Recovery and Utilization of Palm Oil Mill Effluent Source as Value-Added Food Products.

The environmental impacts of palm oil mill effluent (POME) have been a concern due to the water pollution and greenhouse gases emissions. Thus, this study was conducted to recover the value-added products from POME source before being discharged. The samples, before (X) and after (Y) the pre-recovery system in the clarification tank were sampled and analysed and proximate analysis indicated that both samples are rich source of food due to high contents of fats and carbohydrates. GCMS analysis showed that the oil extracts contain predominantly , oleic, linoleic and stearic acids. Regiospecific analysis of oil extracts by quantitative C-NMR spectroscopy demonstrated that both oil extracts contain similar degree of saturation of fatty acids at sn-2 and sn-1,3 positions. The samples are rich in various phytonutrients, pro-vitamin A, vitamin E, squalene and phytosterols, thus contributing to exceptionally high total flavonoid contents and moderate antioxidant activities. Overall, samples X and Y are good alternative food sources, besides reducing the environmental impact of POME.

Keyword: energy

Proteomic effects of repeated-dose oral exposure to 2-monochloropropanediol and its dipalmitate in rat testes.

2- and 3-monochloropropanediol (2-MCPD) and their fatty esters are food contaminants which are concomitantly formed upon thermal treatment of foodstuff containing fats and salt. Exposure to 2- or 3-MCPD thus results, for example, from refined vegetable oils, in instant meals or infant formula, as well as in cereals or pastries. The molecular mechanisms of 2-MCPD toxicity are poorly understood. Here, we performed a comprehensive proteomic analysis of 2-MCDP-induced alterations in the testes from rats following oral administration of 10\u202fmg/kg body weight per day 2-MCPD, or an equimolar dose of 2-MCPD dipalmitate as a representative 2-MCPD fatty ester. In the absence of overt histopathologically detectable toxicity, moderate alterations in cellular proteomic signatures were recorded. The observations are in line with the assumption that the molecular mechanisms of 2-MCPD and 3-MCPD toxicity differ. Observed proteomic alterations point towards effects of 2-MCPD on mitogen-dependent signaling and mitochondrial utilization. Presented data for the first time provide insight into proteomic effects of 2-MCPD in testicular tissue.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Bacterial Microbiota and Metabolic Character of Traditional Sour Cream and Butter in Buryatia, Russia.

Traditional sour cream and butter are widely popular fermented dairy products in Russia for their flavor and nutrition, and contain rich microbial biodiversity, particularly in terms of lactic bacteria (LAB). However, few studies have described the microbial communities and metabolic character of traditional sour cream and butter. The objective of this study was to determine the bacterial microbiota and metabolic character of eight samples collected from herdsmen in Buryatia, Russia. Using single-molecule real-time (SMRT) sequencing techniques, we identified a total of 294 species and/or subspecies in 169 bacterial genera, belonging to 14 phyla. The dominant phylum was Firmicutes (81.47%) and the dominant genus was (59.28%). There were differences between the bacterial compositions of the sour cream and butter samples. The relative abundances of , , and were significantly higher in sour cream than in butter, and the abundance of was significantly lower in sour cream than in butter. Using a pure culture method, 48 strains were isolated and identified to represent seven genera and 15 species and/or subspecies. Among these isolates, subsp. (22.50%) was the dominant LAB species. Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry at elevated was used in combination with statistical methods to detect metabolite differences between traditional sour cream and butter. A total of 27,822 metabolites were detected in all samples, and Lys-Lys, isohexanal, , Leu-Val, and 2\'-deoxycytidine were the most dominant metabolites found in all samples. In addition, 27 significantly different metabolites were detected between the sour cream and butter samples, including short peptides, organic acids, and amino acids. Based on correlation analyses between the most prevalent bacterial species and the main metabolites in sour cream, we conclude that there may be a connection between the dominant LAB species and these metabolites. This study combined omics techniques to analyze the bacterial diversity and metabolic character of traditional sour cream and butter, and we hope that our findings will enrich species resource libraries and provide valuable resources for further research on dairy product flavor.

Keyword: energy

Toxicological evaluation of 2-dodecylcyclobutanone, a unique radiolytic compound of .

This study was conducted to evaluate the toxic effects and potency of 2-dodecylcyclobutanone (2-dDCB), a unique compound derived from via irradiation. In a series of assays of bacterial reverse-mutation, in vitro chromosomal aberration, and in vivo micronucleus, negative responses were found by the treatment of 2-dDCB comparing vehicle control, dimethyl sulfoxide or corn oil. In the acute oral toxicity test, all of the mice administrated 2-dDCB survived, and there were no clinical and necropsy signs observed at any doses (0, 300, and 2000\u202fmg/kg body weight) during the experimental period of 14 days. These results suggested that 2-dDCB is a relatively non-toxic substance with median lethality dose higher than 2000\u202fmg/kg body weight. Moreover, there were no adverse effects noted in rats orally administrated 2-dDCB everyday via gavage for 28 days, even at the highest dose (2.0\u202fmg/kg body weight/day) tested, which is 1000-times higher than the human daily intake of 2-dDCB estimated through an extreme exposure scenario. Overall, these results indicate that 2-dDCB is not likely to raise any human health concerns and irradiated foods containing can be recognized as safe for human consumption under the current international regulation systems for food irradiation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: energy

Rice bran oil ameliorates inflammatory responses by enhancing mitochondrial respiration in murine macrophages.

Previous studies have revealed the anti-inflammatory properties of rice bran oil (RBO), but the detailed mechanisms are poorly understood. Recent studies on the molecular/cellular anti-inflammatory mechanisms of dietary components have demonstrated that mitochondrial respiration plays a key role in macrophage functioning. Since dietary lipids are major substrates for mitochondrial respiration through β-oxidation, the current study examined whether RBO regulates inflammatory responses by modulating mitochondrial metabolism. Palm oil (PO), enriched with which are known to be effectively taken up by cells and used for oxidative phosphorylation, served as a positive control. In the in vitro model of LPS-stimulated RAW 264.7 murine cells, the levels of pro-inflammatory cytokines (IL-6 and TNF-α) in the culture supernatant were significantly reduced by RBO treatment. In contrast, secretion of the anti-inflammatory cytokine IL-10 was upregulated by RBO. Transcription of genes encoding inflammatory mediator molecules (COX-2 and iNOS) and expression of activation markers (CD80, CD86, and MHC-II) in LPS-stimulated RAW 264.7 cells were suppressed by RBO. Mitochondrial respiration (as assessed by an extracellular flux analyzer) increased upon RBO treatment, as the basal respiration, maximal respiration, ATP production, and spare respiratory capacity were upregulated. In an in vivo study, C57BL/6 mice were fed a negative control diet containing corn oil (CO), PO, or RBO for 4 weeks, and bone marrow-derived macrophages (BMDM) were isolated from their tibias and femurs. In pro-inflammatory M1-polarized BMDM (M1-BMDM), the RBO-induced suppression of IL-6 and TNF-α was recapitulated in vivo. Mitochondrial respiration in M1-BMDM also increased following the RBO intervention and the PO control treatment as compared to CO fed negative control. Overall, the current study for the first time demonstrates that RBO regulates inflammatory responses in murine macrophages by upregulating mitochondrial respiration. Further clinical studies are required to validate the animal study.

Keyword: energy

GC-MS Metabolomics Reveals Distinct Profiles of Low- and High-Grade Bladder Cancer Cultured Cells.

Previous studies have shown that metabolomics can be a useful tool to better understand the mechanisms of carcinogenesis; however, alterations in biochemical pathways that lead to bladder cancer (BC) development have hitherto not been fully investigated. In this study, gas chromatography-mass spectrometry (GC-MS)-based metabolomics was applied to unveil the metabolic alterations between low-grade and high-grade BC cultured cell lines. Multivariable analysis revealed a panel of metabolites responsible for the separation between the two tumorigenic cell lines. Significantly lower levels of fatty acids, including myristic, , and palmitoleic acids, were found in high-grade versus low-grade BC cells. Furthermore, significantly altered levels of some amino acids were observed between low- and high-grade BC, namely glycine, leucine, methionine, valine, and aspartic . This study successfully demonstrated the potential of metabolomic analysis to discriminate BC cells according to tumor aggressiveness. Moreover, these findings suggest that bladder tumorigenic cell lines of different grades disclose distinct metabolic profiles, mainly affecting fatty biosynthesis and amino metabolism to compensate for higher energetic needs.

Keyword: energy

High vulnerability of the heart and liver to 3-hydroxypalmitic -induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe liver and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic (3HPA), the long-chain hydroxyl fatty (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca retention capacity and membrane potential in Ca -loaded mitochondria more markedly in the heart and the liver, with mild or no effects in the brain, supporting a higher susceptibility of the heart and the liver to the toxic effects of this fatty . It is postulated that disruption of mitochondrial and Ca homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.© 2018 Wiley Periodicals, Inc.

Keyword: energy

Coupling natural systems with synthetic chemistry for light-driven enzymatic biocatalysis.

Visible light-driven redox reactions have been widely adopted for the production of chemicals to combat shortage and global warming. Key elements of such a reaction system include a photosensitizer, a catalyst, and an electron source. In this review, we introduce the small molecules and nanoparticles that are widely used as photosensitizers, as well as the development of a photosensitizer protein that is based on the expansion of genetic code, with a fluorescent protein that is used as a scaffold. Visible light-driven enzymes using proteins as photosensitizers or as catalysts such as carbon monoxide dehydrogenase (CODH), formic dehydrogenase (FDH), hydrogenase, nitrogenase, cytochrome P450 BM3, and alkane synthase are then described. CODH can be coupled with photosensitizing nanoparticles to reduce CO to CO, and hydrogenase can produce H using high- electrons produced from dye-sensitized nanoparticles. When water-soluble zinc porphyrin is coupled with FDH, visible light drives CO to produce formic . Nitrogenase can reduce N to NH using CdS nanoparticle as photosensitizer. Cytochrome P450 BM3 can be enhanced by a visible light-driven redox system and thus by hydroxylate lauric or fatty acids. CvFAP, an alkane synthase, can decarboxylate to pentadecane under blue light excitation. Moreover, we describe a genetically encoded photosensitive protein, which mimics the function of natural photosynthesis and catalyzes the conversion of CO to CO when covalently attached with a Ni-terpyridine complex.

Keyword: energy

Effect of dietary n-3 polyunsaturated fatty supplementation and post-insemination plane of nutrition on systemic concentrations of metabolic analytes, progesterone, hepatic gene expression and embryo development and survival in beef heifers.

Nutrition, and particularly dietary intake, plays a fundamental role in reproductive function in cattle. There is some evidence that supplemental omega-3 dietary polyunsaturated fatty acids (n-3 PUFA) can exert positive effects on fertility. The objectives of this study were to evaluate the effect of dietary n-3 PUFA supplementation, post-insemination plane of nutrition and their interaction on embryo survival in cattle. Crossbred beef heifers (n\u202f=\u202f185) were individually offered barley straw ad libitum and 6\u202fkg DM of concentrate supplemented with either a rumen-protected source of saturated fatty (; control, CON) or a partially rumen-protected n-3 PUFA-enriched supplement (n-3 PUFA). Estrous was synchronised using two injections of PG administered at 11-d intervals and following artificial insemination (AI\u202f=\u202fDay 0) 179 heifers exhibiting oestrus were inseminated and assigned to one of two dietary treatments: (i) remain on their pre-insemination high dietary plane of nutrition (High) or (ii) restricted to 0.6\xa0× estimated maintenance requirements (Low) in a 2\u202f×\u202f2 factorial design. The heifers were then maintained on their assigned diets until slaughter and embryo recovery on Day 16 (n\u202f=\u202f92) or pregnancy diagnosis by ultrasound scanning at Day 30 post-AI (n\u202f=\u202f87). Plasma concentrations of fatty acids, metabolites, insulin, progesterone (P4) and insulin-like growth factor 1 (IGF-1) were measured at appropriate intervals. Hepatic expression of mRNA for aldo-keto reductase (AKR1C), cytochrome P450 2C (CYP 2C) and cytochrome P450 3A (CYP 3A) was examined. The n-3 PUFA supplementation increased plasma n-3 PUFA concentration (P\u202f<\u202f0.05) and reduced n-6: n-3 PUFA ratio (P\u202f<\u202f0.05). Plasma IGF-1 was higher for n-3 PUFA relative to the CON (P\u202f<\u202f0.05) and for High compared with Low plane of nutrition post-AI (P\u202f<\u202f0.05) groups. A low plane of nutrition post-AI increased plasma concentrations of progesterone from Days 7-16 after insemination (P\u202f<\u202f0.001) but reduced embryo length (P\u202f<\u202f0.001). Supplementation with n-3 PUFA reduced and tended to reduce hepatic expression of CYP2C (P\u202f=\u202f0.01) and CYP3A (P\u202f=\u202f0.08), respectively. However, while dietary n-3 PUFA supplementation and an abrupt reduction in nutrient status following insemination elevated plasma concentrations of n-3 PUFA and mid and late phase P4, respectively, there was no effect of either PUFA supplementation or post-insemination plane of nutrition on embryo survival.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: energy

Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles.

We developed a method that allows for real-time assessment of cellular metabolism in isolated, intact long skeletal muscle fibre bundles from adult mice. This method can be used to study changes in mitochondrial function and fuel utilisation in live skeletal muscle fibre bundles. Our method enables flexibility in experimental design and high-throughput assessment of mitochondrial parameters in isolated skeletal muscle fibre bundles. Extensor digitorum longus (EDL) fibre bundles obtained from chronic high-fat diet fed mice had lower basal oxygen consumption under FCCP-induced maximal respiration, when compared to control chow-fed mice. EDL fibre bundles obtained from chronic high-fat diet fed mice had enhanced mitochondrial oxidation capacity under FCCP-induced maximal respiration, when compared to control chow-fed mice.Metabolic dysfunction in skeletal muscle contributes to the aetiology and development of muscle diseases and metabolic diseases. As such, assessment of skeletal muscle cellular bioenergetics provides a powerful means to understand the role of skeletal muscle metabolism in disease and to identify possible therapeutic targets. Here, we developed a method that allows for the real-time assessment of cellular respiration in intact skeletal muscle fibre bundles obtained from the extensor digitorum longus (EDL) muscle of adult mice. Using this method, we assessed the contribution of ATP turnover and proton leak to basal mitochondrial oxygen consumption rate (OCR). Our data demonstrate that the mitochondria in EDL fibres are loosely coupled. Moreover, in the presence of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), we show that palmitate exposure induced comparable peak OCR and higher total OCR in EDL fibre bundles when compared to pyruvate exposure, suggesting that fatty acids might be a more sustainable fuel source for skeletal muscle when mitochondria are driven to maximal respiration. Application of this method to EDL fibre bundles obtained from chronic high-fat diet fed mice revealed lower basal OCR and enhanced mitochondrial oxidation capacity in the presence of FCCP when compared to the chow-diet fed control mice. By using a 96-well microplate format, our method provides a flexible and efficient platform to investigate mitochondrial parameters of intact skeletal muscle fibres obtained from adult mice.© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

Keyword: energy

Comparative analysis of nutrient content and of eggs from different chicken genotypes.

Eggs are important foods in the daily diet of humans and have great biological activity and a high digestibility. Egg yolk is a good source of biologically active substances such as fatty acids, phospholipids, sterols and tocopherols. The eggs of seven chicken genotypes were analyzed for their chemical composition, and a detailed study of the lipids in egg yolk was conducted. composition of the egg yolk and egg albumen was 29.06-30.51\u2009MJ\u2009kg and 19.77-20.93\u2009MJ\u2009kg respectively. Regarding their chemical composition: water ranged from 471.7 to 515.4\xa0g\u2009kg and 878.3-885.9\xa0g\u2009kg ; fat content in dry matter ranged from 607 to 647\u2009g\u2009kg and 6.7-11.6\xa0g\u2009kg ; protein varied from 302 to 331.7\xa0g\u2009kg and 823.6-892.5\xa0g\u2009kg ; ash ranged from 33.7 to 37.7\xa0g\u2009kg and 63.8-74.0\xa0g\u2009kg ; and nitrogen-free extracts ranged from 12.7 to 36.5\xa0g\u2009kg and 35.0-96.2\xa0g\u2009kg . The sterols and phospholipids in the yolk lipids were 16-26\u2009g\u2009kg and 59-127\u2009g\u2009kg . The main fatty acids in the lipids were oleic (39.1-47.3%) and (26.0-35.5%) acids. Cholesterol in the yolk lipids ranged from 15.9 to 25.9\xa0g\u2009kg . Phosphatidylcholine (389-573\u2009g\u2009kg ), phosphatidylethanolamine (219-355\u2009g\u2009kg ) and phosphatidylinositol (112-284\u2009g\u2009kg ) were the main phospholipids. The content of saturated fatty acids in the phospholipids was significantly higher than that in triacylglycerols.Small variations in the chemical composition of eggs from seven different genotypes were observed. Significant differences in the fatty compositions of the main classes of phospholipids and the triacylglycerol fraction were established. © 2019 Society of Chemical Industry.© 2019 Society of Chemical Industry.

Keyword: energy

The protein-sparing effect of α-lipoic in juvenile grass carp, Ctenopharyngodon idellus: effects on lipolysis, fatty β-oxidation and protein synthesis.

To investigate the protein-sparing effect of α-lipoic (LA), experimental fish (initial body weight: 18·99 (sd 1·82) g) were fed on a 0, 600 or 1200 mg/kg α-LA diet for 56 d, and hepatocytes were treated with 20 μm compound C, the inhibitor of AMP kinase α (AMPKα), treated for 30 min before α-LA treatment for 24 h. LA significantly decreased lipid content of the whole body and other tissues (P0·05). Consistent with results from the experiment in vitro, LA activated phosphorylation of AMPKα and notably increased the protein content of adipose TAG lipase in intraperitoneal fat, hepatopancreas and muscle in vivo (P<0·05). Meanwhile, LA significantly up-regulated the mRNA expression of genes involved in fatty β-oxidation in the same three areas, and LA also obviously down-regulated the mRNA expression of genes involved in amino catabolism in muscle (P<0·05). Besides, it was observed that LA significantly activated the mammalian target of rapamycin (mTOR) pathway in muscle of experimental fish (P<0·05). LA could promote lipolysis and fatty β-oxidation via increasing supply from lipid catabolism, and then, it could economise on the protein from production to increase protein deposition in grass carp. Besides, LA might directly promote protein synthesis through activating the mTOR pathway.

Keyword: energy

Recycling of food waste for fuel precursors using an integrated bio-refinery approach.

The main aim was to integrate FW-recycling with cultivation of Rhodotorula glutinis and anaerobic digestion (AD) for bio- and -fuel recovery. Mixed FW was mechanically macerated (Pcon) and hydrolysed (at 250gL water) via chemical (Ch), thermal (Th) and TCh (combined Ch and Th) treatments. Cleared hydrolysates from individual pre-treatment processes were used as culture medium for cultivation of R. glutinis, while the residual solids (RS) were subjected to AD. Pcon cultivation yielded maximal R. glutinis dry weight biomass (5.18gL) and total fatty contents (1.03gg DW). Dominant fatty methyl esters (FAME) were - (C-26%); Stearic - (C-17%) and Oleic acids (C-38%), ideal for bio-diesel production. Highest methane yields (actual ∼0.139mkg volatile solids) were measured from AD of Th-derived RS. Thus integrated FW recycling approaches will be more feasible for generating and economic incentives.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Effects of Stanniocalcin-1 on glucose flux in rat brown adipose tissue.

The present work assesses in\xa0vitro the role of human Stanniocalcin 1 (hSTC-1) in C-glucose metabolism in brown adipose tissue (BAT) from fed rat. In the fed state, hSTC-1 decreases the incorporation of C from glucose into lipids in the rat BAT. The data support the hypothesis that the capacity of the glycerol-3-phosphate (G3P)-generating pathway (glycolysis) from glucose is regulated by hSTC-1, decreasing the adequate supply of G3P needed for fatty esterification and triacylglycerol (TG) storage in BAT. The results also suggest the effect of hSTC-1 on de novo fatty synthesis from pyruvate generated by C-glucose in the glycolysis pathway. In addition, by decreasing lipogenesis, hSTC-1 increased ATP levels and these two factors may decrease BAT thermogenic function. The presence of hSTC-1 in the incubation medium did not alter C-glucose and C-1- oxidation. The uncoupling protein 1 (UCP-1) expression was not altered by hSTC-1 either. In conclusion, hSTC-1 is one of the hormonal factors that control glucose metabolism in BAT in the fed state. The decrease of TG capacity synthesis from C-glucose by hSTC-1 compromises the BAT thermogenic capacity. Furthermore, the increase in ATP levels would inhibit a futile cycle via UCP-1, which dissipates oxidative as heat.Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: energy

The digestible , metabolizable , and net content of dietary fat sources in thirteen- and fifty-kilogram pigs.

The objective was to determine the concentration of a diverse array of dietary fat sources and, from these data, develop regression equations that explain differences based on chemical composition. A total of 120 Genetiporc 6.0 × Genetiporc F25 (PIC, Inc., Hendersonville, TN) individually housed barrows were studied for 56 d. These barrows (initial BW of 9.9 ± 0.6 kg) were randomly allotted to 1 of 15 dietary treatments. Each experimental diet included 95% of a corn-soybean meal basal diet plus 5% either corn starch or 1 of 14 dietary fat sources. The 14 dietary fat sources (animal-vegetable blend, canola oil, choice white grease source A, choice white grease source B, coconut oil, corn oil source A, corn oil source B, fish oil, flaxseed oil, palm oil, poultry fat, soybean oil source A, soybean oil source B, and tallow) were selected to provide a diverse and robust range of unsaturated fatty :SFA ratios (U:S). Pigs were limit-fed experimental diets from d 0 to 10 and from d 46 to 56, providing a 7-d adaption for fecal collection on d 7 to 10 (13 kg BW) and d 53 to 56 (50 kg BW). At 13 kg BW, the average content of the 14 sources was 8.42 Mcal DE/kg, 8.26 Mcal ME/kg, and 7.27 Mcal NE/kg. At 50 kg BW, the average content was 8.45 Mcal DE/kg, 8.28 Mcal ME/kg, and 7.29 Mcal NE/kg. At 13 kg BW, the variation of dietary fat DE content was explained by DE (Mcal/kg) = 9.363 + [0.097 × (FFA, %)] - [0.016 × omega-6:omega-3 fatty acids ratio] - [1.240 × (arachidic , %)] - [5.054 × (insoluble impurities, %)] + [0.014 × (, %)] ( = 0.008, = 0.82). At 50 kg BW, the variation of dietary fat DE content was explained by DE (Mcal/kg) = 8.357 + [0.189 × U:S] - [0.195 × (FFA, %)] - [6.768 × (behenic , %)] + [0.024 × (PUFA, %)] ( = 0.002, = 0.81). In summary, the chemical composition of dietary fat explained a large degree of the variation observed in the content of dietary fat sources at both 13 and 50 kg BW.

Keyword: energy

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty metabolism was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. Lipid distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral lipid metabolic process" and "Regulation of lipid storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body metabolism.

Keyword: energy

Effect of co-culture with Tetragenococcus halophilus on the physiological characterization and transcription profiling of Zygosaccharomyces rouxii.

Zygosaccharomyces rouxii and Tetragenococcus halophilus are widely existed and play vital roles during the manufacture of fermented foods such as soy sauce. The aim of this study was to elucidate the effect of T. halophilus CGMCC 3792 on the physiological characterizations and transcription profiling of Z. rouxii CGMCC 3791. Salt tolerance analysis revealed that co-culture with T. halophilus enhanced the salt tolerance of Z. rouxii during salt stress. Analysis of the volatile compounds revealed that co-culture reduced the level of 1-butanol, improved the level of octanoic which all were produced by T. halophilus and reduced the level of phenylethyl alcohol produced by Z. rouxii. The presence of Z. rouxii decreased the contents of 3,4-dimethylbenzaldehyde and acetic produced by T. halophilus. In addition, co-culture improved the content of benzyl alcohol significantly. Analysis of membrane fatty showed that co-culture improved the content of (C16:0) and stearic (C18:0) acids in cells of Z. rouxii, and reduced the contents of myristic (C14:0), palmitoleic (C16:1) and oleic (C18:1). In order to further explore the interactions between the two strains, RNA-seq technology was used to investigate the effect of co-culture with T. halophilus on the transcription profiling of Z. rouxii. By comparing cells incubated in co-culture group with cells incubated in single-culture group, a total of 967 genes were considered as differentially expressed genes (DEGs). Among the DEGs, 72 genes were up-regulated, while 895 genes were down-regulated. These DEGs took party in various activities in cells of Z. rouxii, and the result showed co-culture with T. halophilus had a positive effect on proteolysis, the attachment of a cell to another cell, extracellular protein accumulation, metabolism, and a negative effect on oxidative phosphorylation, small molecular substances metabolism, DNA replication and repair, and transcription in cells of Z. rouxii. Results presented in this study may contribute to further understand the interactions between Zygosaccharomyces rouxii and Tetragenococcus halophilus.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

[Insulin resistance is an alimentary deficiency of substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: energy

Prolyl Oligopeptidase Inhibition Attenuates Steatosis in the L02 Human Liver Cell Line.

Prolyl oligopeptidase (POP) is a serine endopeptidase that is widely distributed in vivo, particularly in the liver. Significant changes in functional mitochondrial proteins involved with mitochondrial oxidoreductases/transporters and nucleic binding proteins were observed after POP inhibition in the liver, which suggested a role of POP in regulating liver metabolism. Steatosis in nonalcoholic fatty liver disease (NAFLD) is associated with disturbances in lipid and metabolism in hepatocytes. Here, we aimed to study the effect of POP on hepatocyte steatosis.The human liver cell line L02 was used to investigate the biological effects of POP. An in vitro cell model of steatosis was successfully induced with oleic and . L02 cells were also subjected to S17092 (a POP inhibitor) at different concentrations for 24 or 48 h. Ac-SDKP levels and POP activity were measured to assess the rate of inhibition of POP by S17092. The POP gene and protein expression levels were detected using real-time PCR and Western blots, respectively. Oil red O staining was performed and the triglyceride levels in the L02 cells were also measured. Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The expression of genes involved in lipid metabolism was detected using real-time PCR. The effects of POP inhibition on LC3B II were detected by Western blot.Compared with the control, the POP mRNA levels increased by approximately 30%, and the POP protein levels increased by almost 60% in the steatotic L02 cells. After S17092 (0.026~130 μM) incubation for 24 or 48 h, cell proliferation was significantly decreased in the free fatty (FFA)-treated cells at 26-130 μM; however, S17092 did not affect the proliferation of L02 cells after 24 h of incubation with S17092 at 0.026-65 μM without FFA treatment. S17092 treatment (13 and 26 μM) also elicited no significant effect on apoptosis in normal L02 cells, but FFA treatment increased cell apoptosis, which was attenuated by S17092 incubation. S17092 treatment inhibited intracellular POP activity and decreased the AcSDKP level at the concentration of 0.026-26 μM. After treatment with FFA for 24 h, oil red O staining revealed significant lipid accumulation in the cells in the model group compared with the controls; however, lipid accumulation was suppressed after the administration of S17092 (13 and 26 μM). Accordingly, the triglyceride levels in the FFA-treated cells were approximately 5-fold greater than those of the controls and were decreased by approximately 25% and 45% after the administration of S17092 at 13 and 26 μM, respectively. The mRNA levels of FASN, PPAR-γ, and SREBP-1c were higher in the FFA-treated cells than in the normal controls, and all of these levels were significantly inhibited in the presence of S17092 at both 13 and 26 μM. S17092 treatment did not affect LC3B II in the FFA-treated cells compared with FFA treatment alone.The expression of POP increases with hepatocyte steatosis, and POP inhibitors can significantly reduce intracellular lipid accumulation, which might be related to the inhibition of genes involved in lipid synthesis.

Keyword: energy

Acute hypoxic preconditioning prevents -induced cardiomyocyte apoptosis via switching metabolic GLUT4-glucose pathway back to CD36-fatty dependent.

Metabolic syndrome is a risk factor for the development of cardiovascular diseases. Myocardial cell damage leads to an imbalance of metabolism, and many studies have indicated that short-term hypoxia during myocardial cell injury has a protective effect. In our previous animal studies, we found that short-term hypoxia in the heart has a protective effect, but long-term hypoxia increases myocardial cell injury. (PA)-treated H9c2 cardiomyoblasts and neonatal rat ventricle cardiomyocytes were used to simulate hyperlipidemia model, which suppress cluster of differentiation 36 (CD36) and activate glucose transporter type 4 (GLUT4). We exposed the cells to short- and long-term hypoxia and investigated the protective effects of hypoxic preconditioning on PA-induced lipotoxicity in H9c2 cardiomyoblasts and neonatal rat cardiomyocytes. Preconditioning with short-term hypoxia enhanced both CD36 and GLUT4 metabolism pathway protein levels. Expression levels of phospho-PI3K, phospho-Akt, phospho-AMPK, SIRT1, PGC1α, PPARα, CD36, and CPT1β induced by PA was reversed by short-term hypoxia in a time-dependent manner. PA-induced increased GLUT4 membrane protein level was reduced in the cells exposed to short-term hypoxia and si-PKCζ. Short-term hypoxia, resveratrol and si-PKCζ rescue H9c2 cells from apoptosis induced by PA and switch the metabolic pathway from GLUT4 dependent to CD36 dependent. We demonstrate short-term hypoxic preconditioning as a more efficient way as resveratrol in maintaining the metabolism of hearts during hyperlipidemia and can be used as a therapeutic strategy.© 2017 Wiley Periodicals, Inc.

Keyword: energy

Potential of Whole Pequi ( spp.) Fruit-Pulp, Almond, Oil, and Shell-as a Medicinal Food.

Pequi () pulp, the most consumed component of pequi fruit, is one of the richest Brazilian carotenoid sources, and the most important carotenoid food source native to the Cerrado. However, there are considerable differences among pequi species regarding total carotenoids content and carotenoids profile. Camb. pulp presents higher content of total carotenoids than (Aubl.) Pers. Regarding the carotenoids profile, few studies are available in the literature, mainly with Pequi pulp also has high contents of lipids, dietary fiber, zinc, and magnesium, and is source of calcium and polyphenols. Pequi almond presents high , lipid, protein, dietary fiber, and ash contents. Pequi oil (pulp and almond) has high levels of monounsaturated fatty acids, especially oleic , and relatively high contents of saturated fatty acids, mainly . Pequi shell (exocarp and external mesocarp) is the largest component of the fruit and a solid residue of the pequi processing, which is rich in dietary fibers, including soluble fibers, and phenolic compounds, mostly gallic , ellagic , and quercetin. Pulp oil is the pequi byproduct most investigated in studies. Research with pequi pulp oil in animal models has shown antioxidant, anti-inflammatory, cardioprotective, hepatoprotective, antigenotoxic, and anticarcinogenic effects. In humans, there are evidences supporting anti-inflammatory, cardioprotective, and antigenotoxic effects. Studies on carotenoids profile of pequi pulp in different fruit species are recommended, and studies are necessary to better explore the potential health benefits of pequi fruit components, mainly the pequi pulp and shell.

Keyword: energy

Palm oil and human health. Meeting report of NFI: Nutrition Foundation of Italy symposium.

The use of palm oil by the food industry is increasingly criticized, especially in Italy, for its purported negative effects on human health and environment. This paper summarizes the conclusions of a Symposium on this topic, gathered by the Nutrition Foundation of Italy, among experts representing a number of Italian Medical and Nutritional Scientific Societies. Toxicological and environmental issues were not considered. Participants agreed that: no evidence does exist on the specific health effects of palm oil consumption as compared to other saturated fatty acids-rich fats; the stereospecific distribution of saturated fatty acids in the triacylglycerol molecule of palm oil limits their absorption rate and metabolic effects; in agreement with International guidelines, saturated fatty acids intake should be kept\u2009<10% of total , within a balanced diet; within these limits, no effect of palm oil consumption on human health (and specifically on CVD or cancer risk) can be foreseen.

Keyword: energy

Effect of conjugated linoleic and acetate on milk fat synthesis and adipose lipogenesis in lactating dairy cows.

During biohydrogenation-induced milk fat depression (MFD), nutrients are spared from milk fat synthesis and are available for other metabolic uses. Acetate is the major carbon source spared and it may increase lipid synthesis in adipose tissue during MFD. The objective of this study was to compare the effect of trans-10,cis-12 conjugated linoleic (CLA) and the amount of acetate spared during CLA-induced MFD on adipose tissue lipogenesis. Nine multiparous, lactating, ruminally cannulated Holstein cows (244 ± 107 d in milk; 25 ± 8.4 kg of milk/d; mean ± standard deviation) were randomly assigned to treatments in a 3 × 3 Latin square design. Experimental periods were 4 d followed by a 10-d washout. Treatments were control (CON), ruminal infusion of acetate (AC; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), or abomasal infusion of CLA (10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA). Dry matter intake, milk yield, and milk protein yield and percentage were not affected by treatments. Compared with CON, milk fat yield decreased 23% and fat percent decreased 28% in CLA, and milk fat yield increased 20% in AC. Concentration and yield of milk de novo synthesized fatty acids (C16) was increased by CLA, compared with CON. Yield of de novo synthesized fatty acids and was increased by AC, compared with CON. Lipogenesis capacity of adipose tissue explants was decreased 72% by CLA, but was not affected by AC. Acetate oxidation by adipose explants was not affected by treatments. Treatments had no effect on expression of key lipogenic factors, lipogenic enzymes, and leptin; however, expression of fatty binding protein 4 was reduced in CLA compared with CON. Additionally, hormone-sensitive lipase and perilipin 1 were decreased by CLA and acetate. Plasma glucose and glucagon concentrations were not affected by treatments; however, CLA increased nonesterified fatty acids 17.7%, β-hydroxybutyrate 16.1%, and insulin 27.8% compared with CON, and AC increased plasma β-hydroxybutyrate 18%. In conclusion, during CLA-induced MFD in low-producing cow adipose tissue was sensitive to the anti-lipogenic effects of CLA, while spared acetate did not stimulate adipose lipogenesis. However, acetate may play an important role in stimulating lipogenesis and improving status in the mammary gland under normal conditions.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Selection and Characterization of Responsive Patients with an OXPHOS Complex I Defect.

Mitochondrial disorders are genetically and clinically heterogeneous, mainly affecting high -demanding organs due to impaired oxidative phosphorylation (OXPHOS). Currently, effective treatments for OXPHOS defects, with complex I deficiency being the most prevalent, are not available. Yet, clinical practice has shown that some complex I deficient patients benefit from a high-fat or ketogenic diet, but it is unclear how these therapeutic diets influence mitochondrial function and more importantly, which complex I patients could benefit from such treatment. Dietary studies in a complex I deficient patient with exercise intolerance showed increased muscle endurance on a high-fat diet compared to a high-carbohydrate diet. We performed whole-exome sequencing to characterize the genetic defect. A pathogenic homozygous p.G212V missense mutation was identified in the gene, encoding an early assembly factor of complex I. A complementation study in fibroblasts confirmed that the p.G212V mutation caused the complex I deficiency. The mechanism turned out to be an incomplete assembly of the peripheral arm of complex I, leading to a decrease in the amount of mature complex I. The patient clinically improved on a high-fat diet, which was supported by the 25% increase in maximal OXPHOS capacity in TMEM126B defective fibroblast by the saturated fatty , whereas oleic did not have any effect in those fibroblasts. Fibroblasts of other patients with a characterized complex I gene defect were tested in the same way. Patient fibroblasts with complex I defects in NDUFS7 and NDUFAF5 responded to , whereas ACAD9, NDUFA12, and NDUFV2 defects were non-responding. Although the data are too limited to draw a definite conclusion on the mechanism, there is a tendency that protein defects involved in early assembly complexes, improve with , whereas proteins defects involved in late assembly, do not. Our data show at a clinical and biochemical level that a high fat diet can be beneficial for complex I patients and that our cell line assay will be an easy tool for the selection of patients, who might potentially benefit from this therapeutic diet.

Keyword: energy

Serum metabolomics study of nutrient metabolic variations in chronic heat-stressed broilers.

To investigate the effects of heat stress on broiler metabolism, we assigned 144 broilers to normal control (NC), heat stress (HS) or pair-fed (PF) groups and then monitored the effects using growth performance, carcass characteristics, biochemical assays and GC-MS-based metabolomics. The up-regulation of cloacal temperature confirmed that our experiment was successful in inducing chronic heat stress. The average daily gain and average daily feed intake of the HS group were significantly lower than those of the NC group, by 28·76 and 18·42 %, respectively (P1 and P<0·05). The greater feed:gain ratio of the HS group was significantly positively correlated with the leg, abdominal fat, subcutaneous fat and intramuscular fat proportions and levels of some free amino acids (proline, l-cysteine, methionine and threonine) but was negatively correlated with breast proportion and levels of some NEFA (stearic , arachidonic , and oleic ). These findings indicated that the heat-stressed broilers were in negative balance and unable to effectively mobilise fat, thereby resulting in protein decomposition, which subsequently affected growth performance and carcass characteristics.

Keyword: energy

Understanding the potency of fatty acids with the amino side chains of bovine β lactoglobulin-A quantum chemical approach.

The present study aims to spotlight on the strength of interaction between different fatty acids with the Bovine-lactoglobulin (LGB) protein side chains along with the crystal water molecules at M062X/def2-QZVP level of theory. To analyse the role of carboxylic and its interaction with side chains and to reveal the significance of carboxylic , it was replaced by Acyl chloride (COCl), Acyl Bromide (COBr) and Acyl-Fluorine (COF) group and COS group. The ligands are linear with a straight and branched chain of carbon atoms, but extended methyl group make the ligand bend resulting in non-planar geometry. The least and highest band gap reveals the conductivity properties of ligands. 3UEW, the is well-built owing to the interaction with the amino side chains Lys 69 and Glu 62 resulting in interaction of -124.98kcal/mol. 3D-NCIplot isosurface map and 2D-NCIplot graph plays a key role to confirm and analyse the occurrence of various non covalent interactions. The overall analysis of the fatty acids implies the fact that depending on the aliphatic chain length, the carboxyl group was capable of positioning favourable binding site.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Co-overexpression of native phospholipid-biosynthetic genes plsX and plsC enhances lipid production in Synechocystis sp. PCC 6803.

The overexpression of native plsX and plsC genes involving in fatty /phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and oxygen evolution rate of all overexpressing (OX) strains were normally similar as wild type. For fatty compositions, saturated fatty , in particular (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated fatty acids were observed, specifically linoleic (18:2) and alpha-linolenic (18:3). The plsC/plsX-overexpressing (OX\u2009+\u2009XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX\u2009+\u2009XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.

Keyword: energy

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic metabolism.© 2017 American Heart Association, Inc.

Keyword: energy

Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with metabolic syndrome.

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an -restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with metabolic syndrome.The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of metabolic syndrome.After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk -restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

GLP-1 Analogue, Exendin-4, Modulates MAPKs Activity but not the Heat Shock Response in Human HepG2 Cells.

Glucagon-like peptide-1 (GLP-1) analogues reduce ER stress and inflammation in key metabolic organs, including the liver. However, their effects on heat shock response (HSR) and mitogen-activated protein kinases (MAPKs) have not yet been elucidated. In the present study, we investigate whether the GLP-1 analogue, exendin-4, triggers the expression of HSR and increases MAPK activity under metabolic stress.The effects of exendin-4 in the presence or absence of (PA; 400 μm) or glucose (30 mm) in the HepG2 liver cell line are assessed using Western blots, quantitative real-time PCR, and label-free proteomics.Heat shock proteins (HSP60, HSP72, HSP90, and GRP78) and other chaperones are not significantly affected by exendin-4 under the conditions tested. In contrast, the presence of exendin-4 alone increases the MAPK phosphorylation levels (JNK, ERK1/2, and p38). For short incubation periods, in the presence of PA or glucose, treatment with exendin-4 exhibits limited effects but significantly attenuates MAPK phosphorylation after a 24-h incubation. Interestingly, canonical signaling pathways, such as EIF2, ILK, PKA, and Rho, are modulated by exendin-4.Identifying new pathways modulated by GLP-1 analogues will provide further insights into their benefits beyond their currently recognized roles in glycemic control, such as MAPK activity, homeostasis, and body weight decrease.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: energy

The influence of decreased hydraulic retention time on the performance and stability of co-digestion of sewage sludge with grease trap sludge and organic fraction of municipal waste.

The effect of hydraulic retention time ranging from 12 to 20\xa0d on process performance and stability was investigated in two anaerobic completely stirred tank reactors with a working liquid volume equal to 6 litres. The reactors were fed with mixtures containing (on volatile solids basis): 40% of sewage sludge, 30% of organic fraction of municipal waste and 30% of grease trap sludge. The change of hydraulic retention time did not significantly affect process stability. However, methane yields as well as volatile solids removal decreased from 0.54 to 0.47\xa0l per kg of added volatile solids and 65% to 60% respectively, with the decrease of hydraulic retention time. Despite the fact that the best process performance was achieved for hydraulic retention time of 20 days, the obtained results showed that it is also possible to carry out the co-digestion process at shorter hydraulic retention times with good results. Furthermore, gas production rate as well as biogas production at the shortest hydraulic retention time were approximately 46% higher in comparison to results obtained at the longest hydraulic retention time. In this context, the proposed solution seems to be an interesting option, because it provides an unique opportunity for wastewater treatment plants to improve their profitability by enhancing recovery from sludge as well as full utilisation of the existing infrastructure and hence creates a new potential place for alternative treatment of organic industrial waste such as: fat-rich materials or food waste. However, implementation of the solution at wastewater treatment plants is still a big challenge and needs studies including identification of optimal digesting conditions, information about substrate pumping, inhibition thresholds and processing properties. Additionally, due to the characteristics of both co-substrates their introduction to the full-scale digester should be carefully planned due to a potential risk of overloading of the digester. For this reason, a gradual increase of the share of these wastes in the co-digestion mixture is highly recommended, because it will allow for the acclimatization of bacteria as well as prevent overloading. The results of this study show the importance of gradual acclimatization of microorganisms to the changing environmental conditions. It was found that concentration of long chain fatty acids in effluents increased with the reduction of hydraulic retention time, but this phenomenon did not significantly influence the performance and stability of the process probably due to changes hydraulic retention time being gradual. Although for a moderate negative correlation with volatile solids removal was observed.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

GC-MS-Based metabolomics discovers a shared serum metabolic characteristic among three types of epileptic seizures.

Understanding the overall and common metabolic changes of seizures can provide novel clues for their control and prevention. Here, we aim to investigate the global metabolic feature of serum for three types of seizures.We recruited 27 patients who had experienced a seizure within 48h (including 11 who had a generalized seizure, nine who had a generalized seizure secondary to partial seizure and seven who had a partial seizure) and 23 healthy controls. We analyzed the global metabolic changes of serum after seizures using gas chromatography-mass spectrometry-based metabolomics. Based on differential metabolites, the metabolic pathways and their potential to diagnose seizures were analyzed, and metabolic differences among three types of seizures were compared.The metabolic profiles of serum were distinctive between the seizure group and the controls but were not different among the three types of seizures. Compared to the controls, patients with seizures had higher levels of lactate, butanoic , proline and glutamate and lower levels of , linoleic , elaidic , trans-13-octadecenoic , stearic , citrate, cysteine, glutamine, asparagine, and glyceraldehyde in the serum. Furthermore, these differential metabolites had common change trends among the three types of seizures. Related pathophysiological processes reflected by these metabolites are deficit, inflammation, nervous excitation and neurotoxicity. Importantly, transamination inhibition is suspected to occur in seizures. Lactate, glyceraldehyde and trans-13-octadecenoic in serum jointly enabled a precision of 92.9% for diagnosing seizures.There is a common metabolic feature in three types of seizures. Lactate, glyceraldehyde and trans-13-octadecenoic levels jointly enable high-precision seizure diagnosis.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: energy

feeding increases ceramide supply in association with increased milk yield, circulating nonesterified fatty acids, and adipose tissue responsiveness to a glucose challenge.

Reduced insulin action is a key adaptation that facilitates glucose partitioning to the mammary gland for milk synthesis and enhances adipose tissue lipolysis during early lactation. The progressive recovery of insulin sensitivity as cows advance toward late lactation is accompanied by reductions in circulating nonesterified fatty acids (NEFA) and milk yield. Because can promote insulin resistance in monogastrics through sphingolipid ceramide-dependent mechanisms, (C16:0) feeding may enhance milk production by restoring homeorhetic responses. We hypothesized that feeding C16:0 to mid-lactation cows would enhance ceramide supply and ceramide would be positively associated with milk yield. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows were randomly assigned to a sorghum silage-based diet containing no supplemental fat (control; n=10; 138±45 d in milk) or C16:0 at 4% of ration dry matter (PALM; 98% C16:0; n=10; 136±44 d in milk). Blood and milk were collected at routine intervals. Liver and skeletal muscle tissue were biopsied at d 47 of treatment. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. The plasma and tissue concentrations of ceramide and glycosylated ceramide were determined using liquid chromatography coupled with tandem mass spectrometry. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield, -corrected milk, and milk fat yield. The most abundant plasma and tissue sphingolipids detected were C24:0-ceramide, C24:0-monohexosylceramide (GlcCer), and C16:0-lactosylceramide. Plasma concentrations of total ceramide and GlcCer decreased as lactation advanced, and ceramide and GlcCer were elevated in cows fed PALM. feeding increased hepatic ceramide levels, a response not observed in skeletal muscle tissue. Plasma ceramides (e.g., C24:0-ceramide) were positively correlated with plasma NEFA and milk yield, and positively correlated with NEFA levels following a glucose challenge. Our data demonstrate a remodeled plasma and hepatic sphingolipidome in mid-lactation dairy cows fed PALM. The potential involvement in ceramide in homeorhetic nutrient partitioning to support lactation requires further consideration.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Scale-up cultivation enhanced arachidonic accumulation by red microalgae Porphyridium purpureum.

The present study attempts to cultivate Porphyridium purpureum under different scale-up conditions for further development and commercialization of microalgae-derived PUFAs such as ARA and EPA. Different temperatures (25, 30, and 35\xa0°C) and light intensities (70, 165, and 280\xa0μmol/ms) were applied to the 50 L pilot-scale cultivation of P. purpureum in ASW. The cultivation under the light intensity of 280\xa0μmol/ms at 35\xa0°C obtained biomass concentration up to 9.52\xa0g/L, total fatty content to 56.82\xa0mg/g, and ARA content to 22.29\xa0mg/g. While the maximum EPA content of 7.00\xa0mg/g was achieved under the light intensity of 280\xa0μmol/ms at 25\xa0°C and the highest ratio of UFAs to TFAs of 74.66% was also obtained in this trial. Both biomass concentration and TFAs content were improved by increasing light intensity and temperature. Moreover, the ratio of ARA to EPA was enhanced by increasing cultivation temperature under the light intensity of 280\xa0μmol/ms. In contrast with flask culture, the conversion\xa0of linoleic (C18:2) to ARA was enhanced in scale-up culture, leading to more ARA content. Phosphate limitation enhanced the synthesis of lipid and LPUFAs. Moreover, the biomass concentration and biosynthesis of were preferred by sufficient C (NaHCO).

Keyword: energy

Improvement of extracellular lipase production by a newly isolated Yarrowia lipolytica mutant and its application in the biosynthesis of L-ascorbyl palmitate.

Yarrowia lipolytica Wt-11 producing an extracellular lipase was isolated and identified. To improve the lipase production, Y. lipolytica Wt-11 was subjected to low- ion implantation mutation breeding, and a best mutant, Y. lipolytica Mut-96, was obtained after screening. Under the optimal cultivation conditions, the scaled-up production of lipases were performed, and the lipase activity of Y. lipolytica Mut-96 was enhanced nearly 5.5-fold compared with that of Y. lipolytica Wt-11. After fermentation, the lipases were purified, and the characteristics of the purified lipases were studied. The optimum temperatures and pHs for lipases from Wt-11and Mut-96 were 30°C and 8.0, respectively. The purified lipases were stable between pH 7.0 and 8.5 and unstable at temperatures above 40°C. The lipase activities were enhanced by Ca, Ba, Mn, Fe and SDS. The synthesis of L-ascorbyl palmitate via esterification with L-ascorbic and by immobilized lipases from Wt-11 and Mut-96 in organic media was investigated, and the L-ascorbyl palmitate can be respectively produced at levels of 14.8 and 27.5g/L.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Interfacial curvature effects on the monolayer morphology and dynamics of a clinical lung surfactant.

The morphology of surfactant monolayers is typically studied on the planar surface of a Langmuir trough, even though most physiological interfaces are curved at the micrometer scale. Here, we show that, as the radius of a clinical lung surfactant monolayer-covered bubble decreases to ∼100 µm, the monolayer morphology changes from dispersed circular liquid-condensed (LC) domains in a continuous liquid-expanded (LE) matrix to a continuous LC linear mesh separating discontinuous LE domains. The curvature-associated morphological transition cannot be readily explained by current liquid crystal theories based on isotropic domains. It is likely due to the anisotropic bending of the LC phase of the saturated phospholipids that are common to all natural and clinical lung surfactants. This continuous LC linear mesh morphology is also present on bilayer vesicles in solution. Surfactant adsorption and the dilatational modulus are also strongly influenced by the changes in morphology induced by interfacial curvature. The changes in morphology and dynamics may have physiological consequences for lung stability and function as the morphological transition occurs at alveolar dimensions.

Keyword: energy

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: energy

Impact of Brain Fatty Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with fatty signaling. In humans circulating saturated fatty acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of fatty acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic fatty signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain fatty signaling and its contemporaneous effect on liver function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: energy

Energetic and conformational features linked to the monomeric and dimeric states of bovine BLG.

Bovine β-lactoglobulin (BLG) belong to the lipocalin family. This is a group of proteins involved in the binding and transporting of hydrophobic molecules. Experimental and theoretical reports have stated its complex structural behavior in solution, with coupled effects between homodimerization and ligand recognition. Nonetheless, structural evidence at the atomic level about the cause of this coupled effect has not been reported to date. To address this issue microsecond molecular dynamics (MD) simulations were combined with the molecular mechanics generalized Born surface area (MM/GBSA) approach, clustering analysis and principal component analysis (PCA), to explore the conformational complexity of BLG protein-protein self-association and (PLM) or dodecyl sulfate (SDS) ligand recognition in the monomeric and dimeric state. MD simulations, coupled to the MM/GBSA method, revealed that dimerization exerts contrasting effects on the ligand-binding capacity of BLG. Protein dimerization decreases PLM affinity, promoting dimer association. For SDS the dimeric state increases affinity, enhancing dimer dissociation. MD simulations based on PCA revealed that while few differences in the conformational subspace are observed between the free and bound monomer and dimer coupling for PLM, substantial changes are observed between the free and bound monomer and dimer coupling for SDS.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: energy

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Protocatechuic Ameliorated -Induced Oxidative Damage in Endothelial Cells through Activating Endogenous Antioxidant Enzymes via an Adenosine-Monophosphate-Activated-Protein-Kinase-Dependent Pathway.

Protocatechuic (PCA, 3,4-dihydroxybenzoic ), the main metabolite of anthocyanins, is widely distributed in fruits and vegetables and has been reported to possess a strong antioxidant activity. Herein, we aimed to investigate the protective effect of PCA against high (PA)-induced oxidative damage and the underling molecular mechanisms in human umbilical vein endothelial cells (HUVECs). PCA reduced the levels of intracellular reactive oxygen species and malondialdehyde and increased the activities of endogenous antioxidant enzymes, including superoxide dismutase, glutathione peroxidase 1, and heme oxygenase 1 (HO-1). Metabolomic analysis showed that PCA affected numerous metabolites, especially some of which were related with metabolism. PCA also upregulated the phosphorylation of adenosine-monophosphate-activated protein kinase (AMPK) at Thr through activating liver kinase B1 and then promoted the expression of p-Nrf2 and HO-1. Moreover, PCA reversed the decreased expression of peroxisome proliferator-activated receptor γ coactivator 1α and significantly increased the mitochondrial density. Collectively, these results demonstrated that PCA attenuated PA-induced oxidative damage in HUVECs via an AMPK-dependent pathway.

Keyword: energy

[Influence of Fatty Acids on Oxygen Consumption in Isolated Cardiomyocytes of Rats with Ischemic or Diabetic Heart Disease].

one of the reasons of violation of the functional viability of the myocardium is considered to be the oxygen deprivation and lack of . The reason is the inhibitory effect of fatty acids on glucose oxidation. Recently, however, new data have been published proving the need for fatty acids and their importance in the maintenance and regulation of the functional activity of the myocardium in chronic pathology.to investigate the influence of free polyunsaturated and saturated fatty acids (FA) on the oxygen uptake of isolated cardiomyocytes in intact rats and animals with ischemic or diabetic heart disease.the executed non-randomized controlled study. It includied 3 groups of male rats of Wistar line (weight 250-300g) with 10 animals in each group. Myocardial infarction ("heart attack" group) was caused by ligation of the left coronary artery, diabetes ("diabetes" group)--by intraperitoneal injection of streptozotocin, and "control" group (intact animals). Myocardial infarction caused by ligation of the left coronary artery, and diabetes by intraperitoneal injection of streptozotocin. Isolated cardiac myocytes were obtained by the enzymatic method. Oxygen consumption was assessed polarographically at different saturation incubation medium with oxygen ([O₂] ≤ 8 mg/l and ([O₂] ≥ 16 mg/l). Arachidonic and acids were applied as fatty acids.It is established that the introduction of the incubation medium 20 µm arachidonic or fatty significantly increased the oxygen consumption of intact cardiomyocytes of rats. Both at the ischemic and at the diabetic injury to the heart the opposite result was obtained. The most pronounced decrease in oxygen consumption was indicated in the group with diabetes mellitus.The inhibitory effect of LCD on the rate of oxygen consumption may be associated with the influence of the ischemic or diabetic injury to the heart on the barrierfunction ofmitochondrial membranes of cardiomyocytes, the activity of membrane-associated enzymes and their associated processes.

Keyword: energy

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high- metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: energy

Proximate composition and fatty analysis of (L.) legume seed: implicates to both protein and essential fatty supplementation.

The high mortality rate in Bangladesh is related to poverty, which results in protein malnutrition, essential fatty deficiency and lacks in adequate vitamins, minerals and calorie. Exploring new food items with improved dietary nutrition factors may, therefore, help to decrease the mortality rate in the poor countries like Bangladesh. Accordingly, the present study was a proximate composition and fatty analysis of seed-a legume seed which is given no careful attention locally, though it might be a good source of valuable nutrition factors for both animals and humans. The purpose of the study was, therefore, to generate awareness that could also act as a good source of food components essential for good health. Proximate analysis revealed that the seed powder contained 8.47\xa0±\xa00.52% moisture; 3.50\xa0±\xa00.0.07% ash; 1.02\xa0±\xa00.06% total fat; 23.95\xa0±\xa00.15% total protein; 1.21\xa0±\xa00.16% total dietary fiber; 61.86\xa0±\xa00.70% total carbohydrate and 352.4\xa0±\xa02.66\xa0kcal/100\xa0g . Phytic content (%) was 1.014\xa0±\xa00.048. Major fatty composition (%): the essential fatty linoleic (C18:2, ω-6) was 9.50\xa0±\xa00.68, while the linolenic (C18:3, ω-3) was 1.95\xa0±\xa00.18. (C16:0), stearic (C18:0) and oleic (C18:1) were, respectively, 2.96\xa0±\xa00.19, 0.77\xa0±\xa00.04 and 1.10\xa0±\xa00.06. Lignoceric (C24:0) was 0.11\xa0±\xa00.007%. Monounsaturated palmitoleic (0.006\xa0±\xa00.0), docosapentaenoic (DPA, C22:5, ω-3) and nervonic (0.002\xa0±\xa00.0) were present in trace amounts. Arachidonic (AA, C20:4, ω-6), eicosapentaenoic (C20:5, ω-3), and docosahexaenoic (C22:6, ω-3) were not detected. The fatty profile, thus, suggests that essential omega-6 fatty linoleic (C18:3, ω-6) and omega-3 linolenic (C18:3, ω-3) were the major polyunsaturated fatty acids (PUFA) present in seed. In addition, the seed contained high amount of proteins. Finally, these results suggest that seed could be used as a good source of quality food components, including protein and essential fatty acids.

Keyword: energy

Metabolomic analysis of serum from rats following long-term intake of Chinese sausage.

Owing to the contamination of chemical pollutants, especially nitrosamines and their precursors, in Chinese sausage, long-term intake of Chinese sausage may have potential health effects.This study investigated the effects of long-term intake of Chinese sausage with different contaminations of -nitrosodimethylamine (NDMA) on rat liver and the potential biomarkers in the serum.Serum metabolomic analysis was performed by gas chromatography-mass spectrometry at weeks 7, 17, 25, and 33; simultaneously, liver histopathological examination was conducted and its relationship with the serum metabolomics was also investigated.In the study, long-term intake of Chinese sausage with different NDMA contents induced significant changes in serum metabolites and liver histopathology in rats. Metabonomic analysis showed that seven metabolites - β-alanine, 3-aminoisobutyric , aminooxyacetic , D-alanyl-D-alanine, pelargonic , (PA), and linoleic (LA) - in three sausage diet groups were significantly decreased at four time points, where three other metabolites were notably increased, which included putrescine, ethanolamine phosphate, and taurine. Among the various treatments, the NDMA (sausage-free) group demonstrated the most remarkable changes. Phenylalanine was decreased followed by an increase, and tyrosine persistently declined, both of which were elevated in the NDMA group. In addition, the histopathological result was consistent with that of the serum metabolomic analysis, and the changes in serum metabolites in each sausage diet group and the NDMA group were consistently associated with disorders of lipids, amino , and metabolism.This work indicates that excessive NDMA content in sausage may cause liver damage.

Keyword: energy

Thermo-chemo-sonic pre-digestion of waste activated sludge for yeast cultivation to extract lipids for biodiesel production.

The low cost biosynthesis of microbial lipids are an efficient feedstock to replace plant based oil for biodiesel production. The present study objective is to explore the effect of thermo-chemo-sonic pre-digestion of municipal Waste Activated Sludge (WAS) to cultivate oleaginous L.\xa0starkeyi MTCC-1400 as a model organism to produce high yield biomass and lipid. Higher Suspended Solids (SS) reduction (20 and 15.71%) and Chemical Oxygen Demand (COD) solubilization (27.6 and 22.3%) were achieved at a Specific (SE) input of 5569\xa0kJ/kg for WAS digested with NaOH and KOH, respectively. The maximum biomass of 17.52\xa0g\xa0L and lipid 64.3% dwt were attained in NaOH pre-digested sample. The analyzed lipid profile exhibited high content of (45.6%) and oleic (38.7%) which are more suitable for biofuel production. Thus, these results strongly motivate the use of pre-digested WAS as an efficient and economical substrate for biodiesel production.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Valorization of spent coffee grounds recycling as a potential alternative fuel resource in Turkey: An experimental study.

In this study, recycling of spent coffee grounds (SCG) as a potential feedstock for alternative fuel production and compounds of added value in Turkey was assessed. The average oil content was found (≈ 13% w/w). All samples (before and after extraction) were tested for scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), calorific value, surface analysis and porosity, Fourier transform infrared (FT-IR), and elemental analysis to assess their potential towards fuel properties. Elemental analysis indicated that carbon represents the highest percentages (49.59% and 46.42%, respectively), followed by nitrogen (16.7% and 15.5%), hydrogen (6.74% and 6.04%), and sulfur (0.851% and 0.561%). These results indicate that SCG can be utilized as compost, as it is rich in nitrogen. Properties of the extracted oil were examined, followed by biodiesel production. The quality of biodiesel was compared with American Society for Testing and Materials (ASTM) D6751 standards, and all the properties complied with standard specifications. The fatty compositions were analyzed by gas chromatography. It was observed that coffee waste methyl ester (CWME) is mainly composed of (35.8%) and arachidic (44.6%) acids, which are saturated fatty acids. The low degree of unsaturation provides an excellent oxidation stability (10.4\xa0hr). CWME has also excellent cetane number, higher heating value, and iodine value with poor cold flow properties. The studies also investigated blending of biodiesel with Euro diesel and butanol. Following this, a remarkable improvement in cloud and pour points of biodiesel was obtained. Spent coffee grounds after oil extraction is an ideal material for garden fertilizer, feedstock for ethanol, biogas production, and as fuel pellets. The outcome of such research work produces valuable insights on the recycling importance of SCG in Turkey.Coffee is a huge industry, and coffee has been widely used due to its refreshing properties. This industry generates large quantities of waste. Therefore, recycling of spent coffee grounds for producing alternative fuels and compounds of added value is crucial. Elemental analysis indicated that coffee waste can be utilized as compost, as it is rich in nitrogen. Coffee waste after oil extraction is an ideal feedstock for ethanol and biogas production, garden fertilizer, and as fuel pellets. The low degree of unsaturation provides excellent oxidation stability. Its biodiesel has also excellent cetane number, higher heating value, and lower iodine value.

Keyword: energy

[The physical chemical and biological features of triglycerides. The cell absorption of functionally different +oleic lipoproteins of very low and density and linoleic+linolenic lipoproteins of low density.]

The earlier insulin-independent low-density lipoproteins and more late insulin-dependent very low-density lipoproteins implement different functions at the stages of phylogenesis. The disorder of biological function of trophology, alteration of fatty acids in triglycerides, prevalence of very low-density lipoproteins over oleic very low-density lipoproteins supply mitochondria of cells with non-optimal substrate - saturated fatty for gaining , ATP synthesis. Physiologically, cells implement oleic alternative of fatty acids metabolism, oxidizing mainly ω-9 endogenous oleic mono-unsaturated fatty . The pathology of low density lipoproteins is primary deficiency of poly-unsaturated fatty acids in cells, atherosclerosis and atheromotosis of intima of arteries of elastic type with development of dense plaques from poly-unsaturated fatty acids in the form of polyethers of cholesterol. The pathology of very low-density lipoproteins includes: a) syndrome of resistance to insulin; b) pathology of phylogenetically earlier insulin-independent visceral fatty tissue - metabolic syndrome; c) pathology of phylogenetically later insulin-dependent subcutaneous adipocytes - obesity; d) secondary atherosclerosis, under cumulation of low-density lipoproteins in blood with development of atherothrombosis of intima of arteries, soft plaques rich with triglycerides. As for the prevention of disorders of transfer of fatty acids to very low-density lipoproteins and low-density lipoproteins is common in many ways - minimization of aphysiological effect of surplus amount of food, biological function of diet. The prevention at the level of population includes: a) maximal limitation of content of saturated fatty in food; b) moderate increasing of polysaturated fatty acids, ω-3 poly-saturated fatty acids predominantly; c) increasing of physical activity. The pharmaceuticals are not provided by biology in primary prevention of metabolic pandemics under aphysiological impact of environment factors.

Keyword: energy

Production of 6-O-L-Ascorbyl Palmitate by Immobilized Candida antarctica Lipase B.

L-ascorbyl palmitate (ASP) is an oil-soluble derivative of ascorbic which is used extensively in food, cosmetics industry, and medical hygiene. Enzymatic synthesis of ascorbyl palmitate in tert-butyl alcohol was carried out using indigenously immobilized lipase preparation PyCal with ascorbic and as starting material. The developed batch process under optimized reaction conditions resulted in conversion of 90% with relatively shorter reaction time of 6\xa0h. Continuous process in packed bed reactor gave conversion of 50% with space time yield of 15.46\xa0g/L/h which was found to be higher than the reported literature on enzymatic synthesis of ascorbyl palmitate. The immobilized lipase used in the present work showed good reusability. Characterization of formed ascorbyl palmitate was carried out by FTIR, MS/MS, H-NMR, and C-NMR. The enzymatic process resulted in selective synthesis of 6-O-L-ascorbyl palmitate with purity of 98.6% and no side product formation. The use of underivatized starting materials, high space time yield of 15.46\xa0g\xa0L\xa0h, high recyclability of catalyst, and no by-product formation make the overall process highly efficient and clean in terms of consumption and waste generation, respectively. The optimized reaction parameters for ascorbyl palmitate synthesis in the present study can be used as a useful reference for industrial synthesis of fatty esters of ascorbic by enzymatic route.

Keyword: energy

[Exploration about the protection mechanism of 5-hydroxy-1-methylhydantoin on paraquat poisoning model].

To investigate the effects of 5-hydroxy-1-methylhydantoin (HMH) on kidney injury induced by paraquat (PQ).Fifteen SPF healthy Kunming mice were randomly divided into normal saline (NS) control group, PQ poisoning model group and HMH intervention group, with 5 mice in each group. PQ poisoning model was challenged by one-time gavage of 30 mg/kg PQ solution. The NS group received the same amount of NS by gavage. The HMH group was given 100 mg/kg of HMH immediately after the model was made and continued to be gavaged. Mice in each group were sacrificed 1 day after HMH gavage and heart blood and renal tissue were harvested for examination. The morphological changes of renal tissue were observed under light microscope by hematoxylin-eosin (HE) staining. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in renal tissue were detected according to the instructions of the kit. The expression of heme oxygenase-1 (HO-1) and interleukin-1β (IL-1β) in renal tissues were detected by Western Blot. The serum metabolites were detected by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), the overall distribution of each sample was observed by principal component analysis (PCA), the accuracy of the model was evaluated by multidimensional analysis orthogonal partial least squares-discriminant analysis (OPLS-DA), and the difference metabolites were screened by variable importance in the projection (VIP) value > 1.Light microscopic observation showed that: glomerular structure in NS group was clear, there was no hyperemia and inflammatory cell infiltration in renal interstitium and blood vessels. In PQ group, some glomeruli atrophy and necrosis, capillary congestion in glomeruli, infiltration of inflammatory cells around glomeruli, swelling of renal tubular epithelial cells, slight stenosis of lumen, and occasional necrosis and exfoliation of epithelial cells occurred. The degree of kidney injury in HMH group was significantly less than that in PQ group. Compared with the NS group, the content of MDA in the PQ group was significantly increased (nmol/g: 6.70±0.84 vs. 2.70±0.43, P < 0.01) and the activity of SOD was significantly decreased (kU/L: 33.30±4.66 vs. 50.20±3.23, P < 0.05), the protein expression of HO-1 and IL-1β were significantly increased (HO-1/β-actin: 1.11±0.12 vs. 0.61±0.13, IL-1β/β-actin: 0.93±0.13 vs. 0.32±0.06, both P < 0.05). Compared with the PQ group, the content of MDA in the HMH group was significantly decreased (nmol/g: 5.10±0.93 vs. 6.70±0.84, P < 0.05) and the activity of SOD was significantly increased (kU/L: 61.00±9.02 vs. 33.30±4.66, P < 0.05), the protein expression of HO-1 was significantly decreased (HO-1/β-actin: 0.77±0.07 vs. 1.11±0.12, P < 0.05), however, there was no significant difference in the protein expression of IL-1β (IL-1β/β-actin: 0.87±0.13 vs. 0.93±0.13, P > 0.05). Metabolite detection results showed that: compared with NS group, the levels of creatinine, glycine, succinic , fumaric and citric were significantly increased in the PQ group (VIP value was 1.50, 1.58, 1.64, 1.74 and 1.95 respectively, all P < 0.05), while the levels of ,α-tocopherol and 6-phosphogluconic were significantly decreased (VIP value was 1.10, 1.55 and 1.56 respectively, all P < 0.05). Compared with the PQ group, the levels of creatinine and citric were significantly decreased in the HMH group (VIP value was 1.50 and 1.86, both P < 0.05), while trans-4-hydroxy-proline, D-glyceric , 2, 6-fructose phosphate, 6-phosphate gluconic and aminomalonic were significantly increased (VIP value was 1.36, 1.55, 1.63, 1.68 and 1.76 respectively, all P < 0.05).HMH protects kidney injury caused by PQ poisoning by correcting tricarboxylic acids cycle disturbance, lipid peroxidation and energy metabolism disturbance, and its mechanism is related to the regulation of HO-1 protein expression through Nrf2 pathway.

Keyword: energy

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on lipid metabolism are still poorly understood. Therefore, this study evaluated the fatty (FA) profile and the effects of baru oil on liver and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a lipid emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and liver. The results demonstrated that animals developed marked hypercholesterolemia, liver steatosis, and increased lipid peroxidation in the aorta. Treatment with baru oil attenuated lipid peroxidation and drastically reduced liver damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: energy

Fatty profile of blood plasma and oviduct and uterine fluid during early and late luteal phase in the horse.

During early pregnancy, the secretome of both oviduct and uterus serves as exchange medium for signaling factors between embryo and mother and provides the embryo with nutrients. The preimplantation embryo can utilize the fatty acids (FA) therein via direct incorporation into cell membrane lipid bilayers and for production via β-oxidation. The FA concentration and composition of the oviduct (OF) and uterine fluid (UF) might be regulated by ovarian hormones to meet the changing needs of the growing embryo. In our study, we analyzed the FA profile of blood plasma (BP) and reproductive fluid samples obtained post mortem from slaughtered mares by gas chromatography mass spectrometry. Cycle stage was determined by visual evaluation of the ovary and measurement of plasma progesterone. No major effect of cycle stage on the FA profile was observed. However, the composition of FA was different between BP and both OF and UF. While linoleic, stearic, oleic and were the four most prevalent FA in both BP and reproductive fluids, the latter contained higher concentrations of arachidonic, eicosapentaenoic and dihomo-γ-linolenic . The finding suggests selective endometrial transport mechanisms from plasma into the reproductive fluids or increased endometrial synthesis of selected FA.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: energy

Optimisation, experimental validation and thermodynamic study of the sequential oil extraction and biodiesel production processes from seeds of Sterculia foetida.

Non-edible seeds are not used in any commercial applications, which implies that they can be used for biofuel applications. The present study aimed to maximise the process conditions for oil extraction and sterculia biodiesel production from Sterculia foetida (poon oil). GC-MS identified the methyl esters of sterculia oil as sterculic (32%), (15.88%), oleic (10.00%), linoleic (9.95%) and malvalic (9%). Response surface methodology (RSM) based parametric optimisation of oil extraction was carried out by choosing process variables such as sample weight, volume of solvent to seed ratio and time. The optimum sample amount of 7.5 g and the volume of solvent to seed ratio of 40 mL/g resulted in a maximum oil yield of 45.27% at 3 h. The results were statistically significant (P < 0.05) with a regression coefficient (R) of 0.9988. Furthermore, the artificial neural network (ANN) resulted in an R value greater than 0.9, which validates the RSM. Conventional optimisation of the temperature (55 °C), feedstock to methanol ratio (1:12), catalyst proportion (1.5%) and transesterification reaction time (60 min) yield 90.87% biodiesel production. The physicochemical characteristics of oil and biodiesel complied with the requirements of the ASTM standards. The rate constant and thermodynamic variables at the optimum temperature (333 K) were calculated from the experimental data. The activation (E), activation enthalpy in transition state theory (ΔH), activation entropy in transition state theory (ΔS) and Gibbs free in transition state theory (ΔG) were 37.91 kJ mol, 35.14 kJ mol, - 239.58 J mol K and 79.81 kJ mol respectively. Graphical abstract.

Keyword: energy

Increasing intake enhances milk production and prevents glucose-stimulated fatty disappearance without modifying systemic glucose tolerance in mid-lactation dairy cows.

Feeding saturated fatty acids may enhance milk yield in part by decreasing insulin sensitivity and shifting glucose utilization toward the mammary gland. Our objective was to evaluate the effects of (C16:0) on milk production and insulin sensitivity in cows. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows received a common sorghum silage-based diet and were randomly assigned to a diet containing no supplemental fat (control; n=10; 138±45d in milk) or C16:0 at 4% of ration DM (PALM; 98% C16:0; n=10; 136±44d in milk). Blood and milk were collected at routine intervals. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield by wk 7. Furthermore, PALM increased milk fat yield and -corrected milk at wk 3 and 7. Changes in milk production occurred in parallel with enhanced intake. Increased milk fat yield during PALM treatment was due to increased C16:0 and C16:1 incorporation; PALM had no effect on concentration of milk components, BW, or body condition score. Two weeks posttreatment, -corrected milk and milk fat yield remained elevated in PALM-fed cows whereas yields of milk were similar between treatments. Increased milk fat yield after PALM treatment was due to increased de novo lipogenesis and uptake of preformed fatty acids. The basal concentration of nonesterified fatty acids (NEFA) in plasma increased by d 4, 6, and 8 of PALM treatment, a response not observed thereafter. Although PALM supplementation did not modify insulin, glucose, or triacylglycerol levels in plasma, total cholesterol in plasma was elevated by wk 3. Estimated insulin sensitivity was lower during the first week of PALM treatment; however, glucose disposal following glucose tolerance tests was not modified. In contrast, C16:0 feeding reduced glucose-stimulated NEFA disappearance by wk 7. Results demonstrate that increasing dietary from C16:0 for 7wk improves milk yield and milk composition without modifying systemic glucose tolerance. Reduced glucose-stimulated NEFA disappearance with C16:0 feeding and elevated circulating NEFA may reflect changes in adipose tissue insulin sensitivity.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Consumption of individual saturated fatty acids and the risk of myocardial infarction in a UK and a Danish cohort.

The effect of individual saturated fatty acids (SFAs) on serum cholesterol levels depends on their carbon-chain length. Whether the association with myocardial infarction (MI) also differs across individual SFAs is unclear. We examined the association between consumption of individual SFAs, differing in chain lengths ranging from 4 through 18 carbons, and risk of MI.We used data from 22,050 and 53,375 participants from EPIC-Norfolk (UK) and EPIC-Denmark, respectively. Baseline SFA intakes were assessed through validated, country-specific food frequency questionnaires. Cox regression analysis was used to estimate associations between intakes of individual SFAs and MI risk, for each cohort separately.During median follow-up times of 18.8\u202fyears in EPIC-Norfolk and 13.6\u202fyears in Denmark, respectively, 1204 and 2260 MI events occurred. Mean (±SD) total SFA intake was 13.3 (±3.5) en% in EPIC-Norfolk, and 12.5 (±2.6) en% in EPIC-Denmark. After multivariable adjustment, intakes of C12:0 (lauric ) and C14:0 (myristic ) inversely associated with MI risk in EPIC-Denmark (HR upper versus lowest quintile: 0.80 (95%CI: 0.66, 0.96) for both SFAs). Intakes in the third and fourth quintiles of C4:0-C10:0 also associated with lower MI risk in EPIC-Denmark. Moreover, substitution of C16:0 () and C18:0 (stearic ) with plant proteins resulted in a reduction of MI risk in EPIC-Denmark (HR per 1 %: 0.86 (95%CI: 0.78, 0.95) and 0.87 (95%CI: 0.79, 0.96) respectively). No such associations were found in EPIC-Norfolk.The results from the present study suggest that the association between SFA and MI risk depends on the carbon chain-length of the SFA.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: energy

Isolation and Proteomic Analysis of a Mutant with Enhanced Lipid Production by the Gamma Irradiation Method.

In this study, an enhanced lipid-producing mutant strain of the microalga was developed by gamma irradiation. To induce the mutation, was gamma irradiated at a dose of 400 Gy. After irradiation, the surviving cells were stained with Nile red. The mutant (Cr-4013) accumulating 20% more lipid than the wild type was selected. Thin-layer chromatography revealed the triglyceride and free fatty contents to be markedly increased in Cr-4013. The major fatty acids identified were , oleic , linoleic , and linolenic . Random amplified polymeric DNA analysis showed partial genetic modifications in Cr-4013. To ascertain the changes of protein expression in the mutant strain, two-dimensional electrophoresis was conducted. These results showed that gamma radiation could be used for the development of efficient microalgal strains for lipid production.

Keyword: energy

Exploring Strategies for Labeling Viruses with Gold Nanoclusters through Non-equilibrium Molecular Dynamics Simulations.

Biocompatible gold nanoclusters can be utilized as contrast agents in virus imaging. The labeling of viruses can be achieved noncovalently but site-specifically by linking the cluster to the hydrophobic pocket of a virus via a lipid-like pocket factor. We have estimated the binding affinities of three different pocket factors of echovirus 1 (EV1) in molecular dynamics simulations combined with non-equilibrium free- calculations. We have also studied the effects on binding affinities with a pocket factor linked to the AupMBA nanocluster in different protonation states. Although the absolute binding affinities are over-estimated for all the systems, the trend is in agreement with recent experiments.3 Our results suggest that the natural pocket factor () can be replaced by molecules pleconaril (drug) and its derivative Kirtan1 that have higher estimated binding affinities. Our results also suggest that including the gold nanocluster does not decrease the affinity of the pocket factor to the virus, but the affinity is sensitive to the protonation state of the nanocluster, i.e., to pH conditions. The methodology introduced in this work helps in the design of optimal strategies for gold-virus bioconjugation for virus detection and manipulation.

Keyword: energy

: Physiological Role, Metabolism and Nutritional Implications.

(PA) has been for long time negatively depicted for its putative detrimental health effects, shadowing its multiple crucial physiological activities. PA is the most common saturated fatty accounting for 20-30% of total fatty acids in the human body and can be provided in the diet or synthesized endogenously via lipogenesis (DNL). PA tissue content seems to be controlled around a well-defined concentration, and changes in its intake do not influence significantly its tissue concentration because the exogenous source is counterbalanced by PA endogenous biosynthesis. Particular physiopathological conditions and nutritional factors may strongly induce DNL, resulting in increased tissue content of PA and disrupted homeostatic control of its tissue concentration. The tight homeostatic control of PA tissue concentration is likely related to its fundamental physiological role to guarantee membrane physical properties but also to consent protein palmitoylation, palmitoylethanolamide (PEA) biosynthesis, and in the lung an efficient surfactant activity. In order to maintain membrane phospholipids (PL) balance may be crucial an optimal intake of PA in a certain ratio with unsaturated fatty acids, especially PUFAs of both n-6 and n-3 families. However, in presence of other factors such as positive balance, excessive intake of carbohydrates (in particular mono and disaccharides), and a sedentary lifestyle, the mechanisms to maintain a steady state of PA concentration may be disrupted leading to an over accumulation of tissue PA resulting in dyslipidemia, hyperglycemia, increased ectopic fat accumulation and increased inflammatory tone via toll-like receptor 4. It is therefore likely that the controversial data on the association of dietary PA with detrimental health effects, may be related to an excessive imbalance of dietary PA/PUFA ratio which, in certain physiopathological conditions, and in presence of an enhanced DNL, may further accelerate these deleterious effects.

Keyword: energy

Resistance exercise training and in\xa0vitro skeletal muscle oxidative capacity in older adults.

Whether resistance exercise training (RET) improves skeletal muscle substrate oxidative capacity and reduces mitochondrial production of reactive oxygen species in older adults remains unclear. To address this, 19 older males (≥60\xa0years) were randomized to a RET (n\xa0=\xa011) or to a waitlist control group (n\xa0=\xa08) that remained sedentary for 12\xa0weeks. RET was comprised of three upper body and four lower body movements on resistance machines. One set of 8-12 repetitions to failure of each movement was performed on three nonconsecutive days/week. Improvements in chest press and leg press strength were assessed using a three-repetition maximum (3 RM). Body composition was assessed via dual X-ray absorptiometry. Muscle biopsies were obtained from the vastus lateralis muscle at baseline and at both 3\xa0weeks and 12\xa0weeks. Palmitate and pyruvate oxidation rates were measured from the (14)CO2 produced from [1-(14)C] and [U-(14)C] pyruvate, respectively, during incubation of muscle homogenates. PGC-1α, TFAM, and PPARδ levels were quantified using qRT-PCR Citrate synthase (CS) and β-HAD activities were determined spectrophotometrically. Mitochondrial production of reactive oxygen species (ROS) were assessed using the Amplex Red Hydrogen Peroxide/Peroxidase assay. There were no significant changes in body weight or body composition following the intervention. Chest press and leg press strength (3RM) increased ~34% (both P\xa0<\xa00.01) with RET There were no significant changes in pyruvate or fatty oxidation or in the expression of target genes with the intervention. There was a modest increase (P\xa0<\xa00.05) in βHAD activity with RET at 12\xa0weeks but the change in CS enzyme activity was not significant. In addition, there were no significant changes in ROS production in either group following RET Taken together, the findings of this study suggest that 12\xa0weeks of low volume RET does not increase skeletal muscle oxidative capacity or reduce ROS production in older adults.© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Keyword: energy

Dietary modulation of homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular metabolism is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: energy

Variant Amino Residues Alter the Enzyme Activity of Peanut Type 2 Diacylglycerol Acyltransferases.

Diacylglycerol acyltransferase (DGAT) catalyzes the final step in triacylglycerol (TAG) biosynthesis via the acyl-CoA-dependent acylation of diacylglycerol. This reaction is a major control point in the Kennedy pathway for biosynthesis of TAG, which is the most important form of stored metabolic in most oil-producing plants. In this study, Arachis hypogaea type 2 DGAT (AhDGAT2) genes were cloned from the peanut cultivar \'Luhua 14.\' Sequence analysis of 11 different peanut cultivars revealed a gene family of 8 peanut DGAT2 genes (designated AhDGAT2a-h). Sequence alignments revealed 21 nucleotide differences between the eight ORFs, but only six differences result in changes to the predicted amino (AA) sequences. A representative full-length cDNA clone (AhDGAT2a) was characterized in detail. The biochemical effects of altering the AhDGAT2a sequence to include single variable AA residues were tested by mutagenesis and functional complementation assays in transgenic yeast systems. All six mutant variants retained enzyme activity and produced lipid droplets in vivo. The N6D and A26P mutants also displayed increased enzyme activity and/or total cellular fatty (FA) content. N6D mutant mainly increased the content of palmitoleic , and A26P mutant mainly increased the content of . The A26P mutant grew well both in the presence of oleic and C18:2, but the other mutants grew better in the presence of C18:2. AhDGAT2 is expressed in all peanut organs analyzed, with high transcript levels in leaves and flowers. These levels are comparable to that found in immature seeds, where DGAT2 expression is most abundant in other plants. Over-expression of AhDGAT2a in tobacco substantially increased the FA content of transformed tobacco seeds. Expression of AhDGAT2a also altered transcription levels of endogenous tobacco lipid metabolic genes in transgenic tobacco, apparently creating a larger carbon \'sink\' that supports increased FA levels.

Keyword: energy

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: energy

Prepartum fatty supplementation in sheep I. Eicosapentaenoic and docosahexaenoic supplementation do not modify ewe and lamb metabolic status and performance through weaning.

Fatty acids are involved in the regulation of many physiological pathways, including those involved in gene expression and metabolism. Through effects on these pathways, fatty acids may have lifelong impacts on offspring development and metabolism via maternal supplementation. Therefore, our objective was to investigate the impact of supplementing a source of omega-3 fatty acids eicosapentaenoic (EPA) and docosahexaenoic (DHA) during late gestation on productive and metabolic responses of ewes and their offspring. Eighty-four gestating ewes (28 pens) were blocked and randomly assigned to a diet with 0.39% added fat during the last 50 d of gestation (d -0). The fat sources were Ca salts of a fatty distillate (PFAD) or EPA + DHA. After lambing (d 1), all ewes and lambs were placed on the same pasture. The ewes were weighed and BCS was measured on d -50, -20, 30, and 60 (weaning) of the experiment. Blood samples were taken from the ewes on d -50, -20, 1 (lambing), 30, and 60. Milk yield and composition were measured at 30 d postpartum. Lambs were weighed and bled at d 1, 30, and 60, and ADG was calculated. All plasma samples were analyzed for glucose and NEFA. Ghrelin, prostaglandin E metabolites (PGEM), and the prostaglandin D2 metabolite 11β-PGF2α were measured in d -20 ewe samples. Insulin and adropin were measured in lamb samples at d 60. There was no difference on ewe BW (P = 0.48) or BCS (P = 0.55), or plasma concentrations of glucose (P = 0.57), NEFA (P = 0.44), ghrelin (P = 0.36), PGEM (P = 0.32), and 11β-PGF2α (P = 0.86) between ewes supplemented with PFAD or EPA + DHA. Neither milk yield nor its composition was different (P > 0.10) among treatments. Lambs born from ewes supplemented with PFAD or EPA + DHA did not have different BW (P = 0.22), ADG (P = 0.21) or plasma NEFA (P = 0.52), glucose (P = 0.50), insulin (P = 0.59), and adropin (P = 0.72) concentrations. These results suggest that supplementation of EPA and DHA during late gestation did not affect ewe metabolic profile or milk production. Lamb performance and metabolism through weaning were not affected by maternal supplementation with an enriched source of EPA and DHA.

Keyword: energy

[THE EXCESS OF FATTY IN FOOD AS MAIN CAUSE OF LIPOIDOSIS OF INSULIN-DEPENDENT CELLS: SKELETAL MYOCYTES, CARDIO-MYOCYTES, PERIPORTAL HEPATOCYTES, KUPFFER MACROPHAGES AND B-CELLS OF PANCREAS].

In phylogenesis, becoming of biologicalfunctions and biological reactions proceeds with the purpose ofpermanent increasing of "kinetic perfection ". The main role belongs to factors ofphysical, chemical and biological kinetics, their evaluation using systemic approach technique under permanent effect of natural selection. The late-in-phylogenesis insulin, proceeded with, in development of biological function of locomotion, specialization of insulin-dependent cells: skeletal myocytes, syncytium of cardiomyocytes, subcutaneous adipocytes, periportal hepatocytes, Kupffer\'s macrophages and β-cells of islets of pancreas. The insulin initiated formation of new, late in phylogenesis, large pool of fatty cells-subcutaneous adipocytes that increased kinetic parameters of biological function of locomotion. In realization of biological function of locomotion only adipocytes absorb exogenous mono unsaturated and saturated fatty acids in the form of triglycerides in composition of oleic and lipoproteins of very low density using apoE/B-100 endocytosis. The rest of insulin-dependent cells absorb fatty acids in the form of unesterified fatty acids from associates with albumin and under effect of CD36 of translocase offatty acids. The insulin in all insulin-depended cells inhibits biological reaction of lipolysis enhancing contributing into development of lipoidosis. The insulin expresses transfer offatty acids in the form of unsaturated fatty acids from adipocytes into matrix of mitochondria. The insulin supplies insulin-dependent cells with substrates for acquiring subject to that in pool of unsaturated fatty acids in adipocytes prevails hydrophobic unsaturated fatiy that slowly passes into matrix through external membrane ofmitochondria; oxidases of mitochondria so slowly implement its β-oxidation that content of exogenous unsaturatedfatty can\'t be higher than phylogenetic, physiological level - 15% of all amount offatty acids transferring to insulin-dependent cells. The insulin can\'t both to decrease content of exogenous fatty and inhibit lipolysis in visceral fatty cells of omentum.

Keyword: energy

GC-MS metabolomics reveals disturbed metabolic pathways in primary mouse hepatocytes exposed to subtoxic levels of 3,4-methylenedioxymethamphetamine (MDMA).

3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a well-known hepatotoxic drug. Although its toxicity has been thoroughly studied at high concentrations, there is still insufficient knowledge on possible alterations of cell function at subtoxic concentrations, which are in fact more representative concentrations of intoxication scenarios. In this study, a gas chromatography-mass spectrometry (GC-MS) metabolomics approach was used to investigate the metabolic changes in primary mouse hepatocytes (PMH) exposed to two subtoxic concentrations of MDMA (LC and LC) for 24\xa0h. Metabolomic profiling of both intracellular metabolites and volatile metabolites in the extracellular medium of PMH was performed. Multivariate analysis showed that the metabolic pattern of cells exposed to MDMA discriminates from the controls in a concentration-dependent manner. Exposure to LC MDMA induces a significant increase in some intracellular metabolites, including oleic and , and a decrease in glutamate, aspartate, 5-oxoproline, fumarate, malate, phosphoric , α-ketoglutarate and citrate. Extracellular metabolites such as acetophenone, formaldehyde, pivalic , glyoxal and 2-butanone were found significantly increased after exposure to MDMA, compared to controls, whereas 4-methylheptane, 2,4-dimethyl-1-heptene, nonanal, among others, were found significantly decreased. The panel of discriminatory metabolites is mainly involved in tricarboxylic (TCA) cycle, fatty metabolism, glutamate metabolism, antioxidant defenses and possibly changes in the liver enzyme machinery. Overall, these results highlight the potential of the intra- and extracellular metabolome to study alterations triggered by subtoxic concentrations of MDMA in hepatic cell functions, which represents a more realistic appraisal of early toxicity events posed by exposure to this drug. In addition, these results also revealed some metabolites that may be used as potential biomarkers indicative of early events in the hepatotoxicity induced by MDMA.

Keyword: energy

P66Shc-Induced MicroRNA-34a Causes Diabetic Endothelial Dysfunction by Downregulating Sirtuin1.

Diabetes mellitus causes vascular endothelial dysfunction and alters vascular microRNA expression. We investigated whether endothelial microRNA-34a (miR-34a) leads to diabetic vascular dysfunction by targeting endothelial sirtuin1 (Sirt1) and asked whether the oxidative stress protein p66Shc governs miR-34a expression in the diabetic endothelium.MiR-34a is upregulated, and Sirt1 downregulated, in aortic endothelium of db/db and streptozotocin-induced diabetic mice. Systemic administration of miR-34a inhibitor, or endothelium-specific knockout of miR-34a, prevents downregulation of aortic Sirt1 and rescues impaired endothelium-dependent aortic vasorelaxation induced by diabetes mellitus. Moreover, overexpression of Sirt1 mitigates impaired endothelium-dependent vasorelaxation caused by miR-34a mimic ex vivo. Systemic infusion of miR-34a inhibitor or genetic ablation of endothelial miR-34a prevents downregulation of endothelial Sirt1 by high glucose. MiR-34a is upregulated, Sirt1 is downregulated, and oxidative stress (hydrogen peroxide) is induced in endothelial cells incubated with high glucose or the free fatty palmitate in vitro. Increase of hydrogen peroxide and induction of endothelial miR-34a by high glucose or palmitate in vitro is suppressed by knockdown of p66shc. In addition, overexpression of wild-type but not redox-deficient p66Shc upregulates miR-34a in endothelial cells. P66Shc-stimulated upregulation of endothelial miR-34a is suppressed by cell-permeable antioxidants. Finally, mice with global knockdown of p66Shc are protected from diabetes mellitus-induced upregulation of miR-34a and downregulation of Sirt1 in the endothelium.These data show that hyperglycemia and elevated free fatty acids in the diabetic milieu recruit p66Shc to upregulate endothelial miR-34a via an oxidant-sensitive mechanism, which leads to endothelial dysfunction by targeting Sirt1.© 2016 American Heart Association, Inc.

Keyword: energy

Altering the ratio of dietary , stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, partitioning, and production responses of dairy cows.

The objective of this study was to evaluate the effects of varying the ratio of dietary (C16:0), stearic (C18:0), and oleic (cis-9 C18:1) acids in basal diets containing soyhulls or whole cottonseed on nutrient digestibility, partitioning, and production response of lactating dairy cows. Twenty-four mid-lactation multiparous Holstein cows were used in a split-plot Latin square design. Cows were allocated to a main plot receiving either a basal diet with soyhulls (SH, n = 12) or a basal diet with whole cottonseed (CS, n = 12) that was fed throughout the experiment. Within each plot a 4 × 4 Latin square arrangement of treatments was used in 4 consecutive 21-d periods. Treatments were (1) control (CON; no supplemental fat), (2) high C16:0 supplement [PA; fatty (FA) supplement blend provided ∼80% C16:0], (3) C16:0 and C18:0 supplement (PA+SA; FA supplement blend provided ∼40% C16:0 + ∼40% C18:0), and (4) C16:0 and cis-9 C18:1 supplement (PA+OA; FA supplement blend provided ∼45% C16:0 + ∼35% cis-9 C18:1). Interactions between basal diets and FA treatments were observed for dry matter intake (DMI) and milk yield. Among the SH diets, PA and PA+SA increased DMI compared with CON and PA+OA treatments, whereas in the CS diets PA+OA decreased DMI compared with CON. The PA, PA+SA, and PA+OA treatments increased milk yield compared with CON in the SH diets. The CS diets increased milk fat yield compared with the SH diets due to the greater yield of de novo and preformed milk FA. The PA treatment increased milk fat yield compared with CON, PA+SA, and PA+OA due to the greater yield of mixed-source (16-carbon) milk FA. The PA treatment increased 3.5% fat-corrected milk compared with CON and tended to increase it compared with PA+SA and PA+OA. The CS diets increased body weight (BW) change compared with the SH diets. Additionally, PA+OA tended to increase BW change compared with CON and PA and increased it in comparison with PA+SA. The PA and PA+OA treatments increased dry matter and neutral detergent fiber digestibility compared with PA+SA and tended to increase them compared with CON. The PA+SA treatment reduced 16-carbon, 18-carbon, and total FA digestibility compared with the other treatments. The CS diets increased partitioning toward body reserves compared with the SH diets. The PA treatment increased partitioning toward milk compared with CON and PA+OA and tended to increase it compared with PA+SA. In contrast, PA+OA increased partitioned to body reserves compared with PA and PA+SA and tended to increase it compared with CON. In conclusion, milk yield responses to different combinations of FA were affected by the addition of whole cottonseed in the diet. Among the combinations of C16:0, C18:0, and cis-9 C18:1 evaluated, fat supplements with more C16:0 increased output in milk, whereas fat supplements with more cis-9 C18:1 increased storage in BW. The combination of C16:0 and C18:0 reduced nutrient digestibility, which most likely explains the lower performance observed compared with other treatments.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines metabolism, and gut microbiome-derived metabolism. Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for supply, resulting in negative balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested fat decomposition compensating for shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: energy

Palmitoleic (16:1n7) increases oxygen consumption, fatty oxidation and ATP content in white adipocytes.

We have recently demonstrated that palmitoleic (16:1n7) increases lipolysis, glucose uptake and glucose utilization for production in white adipose cells. In the present study, we tested the hypothesis that palmitoleic modulates bioenergetic activity in white adipocytes.For this, 3\xa0T3-L1 pre-adipocytes were differentiated into mature adipocytes in the presence (or absence) of (16:0) or palmitoleic (16:1n7) at 100 or 200\xa0μM. The following parameters were evaluated: lipolysis, lipogenesis, fatty (FA) oxidation, ATP content, oxygen consumption, mitochondrial mass, citrate synthase activity and protein content of mitochondrial oxidative phosphorylation (OXPHOS) complexes.Treatment with 16:1n7 during 9\xa0days raised basal and isoproterenol-stimulated lipolysis, FA incorporation into triacylglycerol (TAG), FA oxidation, oxygen consumption, protein expression of subunits representing OXPHOS complex II, III, and V and intracellular ATP content. These effects were not observed in adipocytes treated with 16:0.Palmitoleic , by concerted action on lipolysis, FA esterification, mitochondrial FA oxidation, oxygen consumption and ATP content, does enhance white adipocyte expenditure and may act as local hormone.

Keyword: energy

Short-term starvation is a strategy to unravel the cellular capacity of oxidizing specific exogenous/endogenous substrates in mitochondria.

Mitochondrial oxidation of nutrients is tightly regulated in response to the cellular environment and changes in demands. studies evaluating the mitochondrial capacity of oxidizing different substrates are important for understanding metabolic shifts in physiological adaptations and pathological conditions, but may be influenced by the nutrients present in the culture medium or by the utilization of endogenous stores. One such influence is exemplified by the Crabtree effect (the glucose-mediated inhibition of mitochondrial respiration) as most experiments are performed in glucose-containing media. Here, using high-resolution respirometry, we evaluated the oxidation of endogenous or exogenous substrates by cell lines harboring different metabolic profiles. We found that a 1-h deprivation of the main energetic nutrients is an appropriate strategy to abolish interference of endogenous or undesirable exogenous substrates with the cellular capacity of oxidizing specific substrates, namely glutamine, pyruvate, glucose, or palmitate, in mitochondria. This approach primed mitochondria to immediately increase their oxygen consumption after the addition of the exogenous nutrients. All starved cells could oxidize exogenous glutamine, whereas the capacity for oxidizing palmitate was limited to human hepatocarcinoma Huh7 cells and to C2C12 mouse myoblasts that differentiated into myotubes. In the presence of exogenous glucose, starvation decreased the Crabtree effect in Huh7 and C2C12 cells and abrogated it in mouse neuroblastoma N2A cells. Interestingly, the fact that the Crabtree effect was observed only for mitochondrial basal respiration but not for the maximum respiratory capacity suggests it is not caused by a direct effect on the electron transport system.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: energy

Increased FNDC5/Irisin expression in human hepatocellular carcinoma.

The fibronectin type III domain containing 5 (FNDC5)/Irisin, a novel -regulating hormone, is associated with lipid and carbohydrate metabolism. It is produced in low amounts by normal hepatic tissue, while in human hepatocellular carcinoma (HCC), in which aberrant de novo lipogenesis (DNL) occurs, the hepatic expression of FNDC5/Irisin is still unknown. The gene expression of FNDC5/Irisin, associated to key regulators of DNL, inflammation and cancer progression was evaluated in liver tissue of 18 patients with HCC undergoing liver transplantation and of 18 deceased donors. Hepatic mRNA expression of FNDC5/Irisin and stearoyl-CoA desaturase (SCD-1), main enzymatic regulator of DNL, were significantly higher in HCC patients than in donors (p<0.0001 and p=0.015, respectively). The hepatic mRNA expression of the neurogenic locus notch homolog protein 1 (NOTCH1) tended to be higher in HCC patients than in donors (p=0.06). Only in HCC patients, hepatic FNDC5/Irisin strongly correlated with the transcription factor sterol regulatory element-binding factor 1, SCD-1, NOTCH1, tumor necrosis factor-α and Interleukin-6 mRNA expression. Further, in HCC patients, FNDC5/Irisin mRNA tended to correlate to plasma lipid profile namely triglycerides, /linoleic and polyunsaturated fatty /saturated fatty ratios. In conclusion, HCC-liver tissue over-expressed FNDC5/Irisin in association with gene expression of mediators involved in lipogenesis, inflammation and cancer, suggesting a possible protective role of the hormone from the liver damage.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: energy

Environmental alterations in biofuel generating molecules in Zilla spinosa.

Now days, production of fuels and petrochemicals from renewable lignocellulosic biomass is an indispensable issue to meet the growing demand. Meanwhile, the changes in the climate and soil topography influence the growth and development as well as canopy level of the lignocellulosic biomass. In this study, Zilla spinosa Turr (Zilla) plants with similar age and size were collected from three main sectors (upstream, midstream, and downstream) of Wadi Hagul during spring (April) and summer (July) seasons. Environmental stresses evoked reduction in the trapping pigments concomitant with increments in chlorophyll fluorescence in summer harvested plants particularly at downstream. Furthermore, the biofuels generating compounds including carbohydrate, lignin, and lipid making the plant biomasses are greatly affected by environmental conditions. Greater amount of lignin was estimated in summer harvested Z. spinosa shoots particularly at downstream. Moreover, the total oil content which is a promising source of biodiesel was considerably decreased during summer season particularly at downstream. The physical properties of the lipids major constituent fatty methyl esters determine the biofuel properties and contribute in the adaptation of plants against environmental stresses. Hence, the analysis of fatty profile showed significant modifications under combined drought and heat stress displayed in the summer season. The maximum increase in saturated fatty levels including tridecanoic (C13:0), pentadeanoic (C15:0), (C16:0), and stearic (C18:0) were estimated in spring harvested Z. spinosa aerial portions particularly at midstream. In spite of the reduction in the total oil content, a marked increase in the value of unsaturated to saturated fatty acids ratio and thereby the unsaturation index were achieved during the dry summer period. Henceforth, these seasonal and spatial variations in fatty acids profiles may contribute in the acclimatization of Z. spinosa plants to soil water scarcity associated with heat stress experienced during summer. In addition, the alterations in the fatty profiles may match biofuel requirements. In conclusion, the most adequate growing season (spring) will be decisive for achieving high lipid productivity associated with improved biofuel quality in terms of high saturated fatty acids percentage that improves its cetane number. However, the dry summer season enhanced the accumulation of greater amount of lignin that may enhance the biodiesel quantity.

Keyword: energy

Chemical composition, in vitro gas production, methane production and fatty profile of canola silage (Brassica napus) with four levels of molasses.

The objective of this study was to investigate the effect of four levels of molasses on chemical composition, in vitro digestibility, methane production and fatty profile of canola silages. A canola (Brassica napus var. Monty) crop was established in a small-scale agricultural farm and harvested 148\xa0days after sowing. Four levels of molasses were tested with respect to the fresh weight (1.5\xa0kg); these were 1% (CS-1), 2% (CS-2), 3% (CS-3) and 4% (CS-4) molasses, and 0% molasses (CS-0) was included as a control. A total of 45 microsilages were prepared using PVC pipes (4\xa0in. of diameter × 20\xa0cm of length), and the forage was compressed using a manual press. The effects of control and treatments were tested using the general linear model Y\u2009=\u2009μ\u2009+\u2009T\u2009+\u2009E. The linolenic (C18:3n3), (C16:0) and linoleic methyl ester (C18:2n6c) accounted for 30%, 21% and 10.5% of total fatty acids, respectively; the fermentation parameters and in vitro methane production were not affected (P\u2009>\u20090.05) by treatments; in vitro digestibility decreased significantly (P\u2009<\u20090.05) as the level of molasses increased. It was concluded that CS-4 improved the DM content by 9% and showed high content of linolenic methyl ester. The gross of canola silages could favour the oleic methyl ester.

Keyword: energy

High α-tocopherol dosing increases lipid metabolism by changing redox state in damaged rat gastric mucosa and liver after ethanol treatment.

Regeneration of ethanol-injured rat gastric mucosa must undergo changes in major metabolic pathways to achieve DNA replication and cell proliferation. These events are highly dependent on glucose utilization and inhibited by vitamin E (VE) (α-tocopherol) administration. Therefore, the present study aimed at assessing lipid metabolism in the gastric mucosa and ethanol-induced gastric damage and the effect of α-tocopherol administration. For this, rates of fatty β-oxidation and lipogenesis were tested in gastric mucosa samples. Through histological analysis, we found loss of the mucosa\'s superficial epithelium, which became gradually normalized during the recovery period. Proliferation of gastric mucosa occurred with augmented formation of β-oxidation by-products, diminished synthesis of triacylglycerols (TGs), as well as of phospholipids, and a reduced cytoplasmic NAD/NADH ratio, whereas the mitochondrial redox NAD/NADH ratio was much less affected. In addition, α-tocopherol increased utilization in the gastric mucosa, which was accompanied by the induction of \'mirror image\' effects on the cell redox state, reflected in an inhibited cell gastric mucosa proliferation by the vitamin administration. In conclusion, the present study shows, for the first time, the role of lipid metabolism in the adaptive cell gastric mucosa changes that drive proliferation after a chronic insult. Moreover, α-tocopherol increased gastric mucosa utilization of associated with production. These events could be associated with its antioxidant properties in co-ordination with regulation of genes and cell pathways, including changes in the cell NAD/NADH redox state.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: energy

Enhanced lipid extraction from oleaginous yeast biomass using ultrasound assisted extraction: A greener and scalable process.

Soaring demand for alternative fuels has been gaining wide interest due to depletion of conventional fuel, increasing petroleum prices and greenhouse gas emissions. Biodiesel, an alternative fuel, derived from oleaginous microbes has been promising because of short incubation time and easy to scale up. Oleaginous yeast Trichosporon sp. is capable of utilizing glycerol and agro-residues for enhanced lipid synthesis. Lipid extraction from Trichosporon sp. biomass showed highest lipid content with ultrasonic assisted extraction (43\u202f±\u202f0.33%, w/w) coupled with process parameters than the conventional Soxhlet (30\u202f±\u202f0.28%, w/w) and Binary solvent [choloroform:methanol, (2:1, v/v)] methods (36\u202f±\u202f0.38%, w/w), respectively. The standardized process parameters of ultrasonic assisted extraction coupled with chloroform/methanol solvent system resulted 95-97% of conversion efficiency in 20\u202fmin at 30\u202f°C with a frequency of 50\u202fHz and 2800\u202fW power, respectively. Enzymatic transesterification of yeast biomass lipid obtained 85% of fatty methyl esters that are predominant with oleic methyl ester followed by and stearic methyl esters, respectively. These results substantiate that the ultrasonic assisted extraction is a potential green extraction technique that had reduced time, energy and solvent consumption without compromising on lipid quality. Deploying this green extraction technique could make the biodiesel production process inexpensive and eco-friendly.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Study of the Thermal Properties and the Fire Performance of Flame Retardant-Organic PCM in Bulk Form.

The implementation of organic phase change materials (PCMs) in several applications such as heating and cooling or building comfort is an important target in thermal storage (TES). However, one of the major drawbacks of organic PCMs implementation is flammability. The addition of flame retardants to PCMs or shape-stabilized PCMs is one of the approaches to address this problem and improve their final deployment in the building material sector. In this study, the most common organic PCM, Paraffin RT-21, and fatty acids mixtures of capric (CA), myristic (MA), and (PA) in bulk, were tested to improve their fire reaction. Several flame retardants, such as ammonium phosphate, melamine phosphate, hydromagnesite, magnesium hydroxide, and aluminum hydroxide, were tested. The properties of the improved PCM with flame retardants were characterized by thermogravimetric analyses (TGA), the dripping test, and differential scanning calorimetry (DSC). The results for the dripping test show that fire retardancy was considerably enhanced by the addition of hydromagnesite (50 wt %) and magnesium hydroxide (50 wt %) in fatty acids mixtures. This will help the final implementation of these enhanced PCMs in building sector. The influence of the addition of flame retardants on the melting enthalpy and temperatures of PCMs has been evaluated.

Keyword: energy

Metabolomic study for monitoring of biomarkers in mouse plasma with asthma by gas chromatography-mass spectrometry.

Asthma is a multifaceted chronic disease caused by an alteration of various genetic and environmental factors that is increasing in incidence worldwide. However, the biochemical mechanisms regarding asthma are not completely understood. Thus, we performed of metabolomic study for understanding of the biochemical events by monitoring of altered metabolism and biomarkers in asthma. In mice plasma, 27 amino acids(AAs), 24 fatty acids(FAs) and 17 organic acids(OAs) were determined by ethoxycarbonyl(EOC)/methoxime(MO)/tert-butyldimethylsilyl(TBDMS) derivatives with GC-MS. Their percentage composition normalized to the corresponding mean levels of control group. They then plotted as star symbol patterns for visual monitoring of altered metabolism, which were characteristic and readily distinguishable in control and asthma groups. The Mann-Whitney test revealed 25 metabolites, including eight AAs, nine FAs and eight OAs, which were significantly different (p<0.05), and orthogonal partial least-squares-discriminant analysis revealed a clear separation of the two groups. In classification analysis, and methionine were the main metabolites for discrimination between asthma and the control followed by pipecolic, lactic, α-ketoglutaric, and linoleic acids for high classification accuracy as potential biomarkers. These explain the metabolic disturbance in asthma for AAs and FAs including intermediate OAs related to the metabolism in the TCA cycle.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Physicochemical properties, nutritional value and techno-functional properties of goldenberry (Physalis peruviana) waste powder concise title: Composition of goldenberry juice waste.

Goldenberry waste powder, contained 5.87% moisture, 15.89% protein, 13.72% fat, 3.52% ash, 16.74% dietary fiber and 61% carbohydrates. Potassium (560\u202fmg/100\u202fg) was the predominant element followed by sodium (170\u202fmg/100\u202fg) and phosphorus (130\u202fmg/100\u202fg). Amino analysis gave high levels of cystine/methionine, histidine and tyrosine/phenylalanine. Goldenberry waste powder had good levels of the techno-functional properties including water absorption index, swelling index, foaming capacity and stability (3.38\u202fg/g, 5.24\u202fml/g, 4.09 and 72.0%, respectively). Fatty acids profile showed that linoleic was the predominant fatty followed by oleic, and stearic acids. Iodine value (109.5\u202fg/100\u202fg of oil), value (2.36\u202fmg KOH/g of oil), saponification value (183.8\u202fmg KOH/g of oil), peroxide value (8.2\u202fmeq/kg of oil) and refractive index (1.4735) were comparable to those of soybean and sunflower oils. Goldenberry waste oil exhibited absorbance in the UV range at 100-400\u202fnm.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Thermodynamic versus non-equilibrium stability of monolayers in calcium-enriched sea spray aerosol proxy systems.

Of the major cations in seawater (Na+, Mg2+, Ca2+, K+), Ca2+ is found to be the most enriched in fine sea spray aerosols (SSA). In this work, we investigate the binding of Ca2+ to the carboxylic headgroup of (PA), a marine-abundant fatty , and the impact such binding has on the stability of PA monolayers in both equilibrium and non-equilibrium systems. A range of Ca2+ conditions from 10 μM to 300 mM was utilized to represent the relative concentration of Ca2+ in high and low relative humidity aerosol environments. The CO2- stretching modes of PA detected by surface-sensitive infrared reflection-absorption spectroscopy (IRRAS) reveal ionic binding motifs of the Ca2+ ion to the carboxylate group with varying degrees of hydration. Surface tensiometry was used to determine the thermodynamic equilibrium spreading pressure (ESP) of PA on the various aqueous CaCl2 subphases. Up to concentrations of 1 mM Ca2+, each system reached equilibrium, and Ca2+:PA surface complexation gave rise to a lower state revealed by elevated surface pressures relative to water. We show that PA films are not thermodynamically stable at marine aerosol-relevant Ca2+ concentrations ([Ca2+] ≥ 10 mM). IRRAS and vibrational sum frequency generation (VSFG) spectroscopy were used to investigate the surface presence of PA on high concentration Ca2+ aqueous subphases. Non-equilibrium relaxation (NER) experiments were also conducted and monitored by Brewster angle microscopy (BAM) to determine the effect of the Ca2+ ions on PA stability. At high surface pressures, the relaxation mechanisms of PA varied among the systems and were dependent on Ca2+ concentration.

Keyword: energy

Proteomic profiling and integrated analysis with transcriptomic data bring new insights in the stress responses of after an arrest during high-temperature ethanol fermentation.

The thermotolerant yeast is a potential candidate for high-temperature fermentation. When was used for high-temperature ethanol fermentation, a fermentation arrest was observed during the late fermentation stage and the stress responses have been investigated based on the integration of RNA-Seq and metabolite data. In order to bring new insights into the cellular responses of after the fermentation arrest during high-temperature ethanol fermentation, quantitative proteomic profiling and integrated analysis with transcriptomic data were performed in this study.Samples collected at 14, 16, 18, 20 and 22\xa0h during high-temperature fermentation were subjected to isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic profiling and integrated analysis with transcriptomic data. The correlations between transcripts and proteins for the comparative group 16\xa0h vs 14\xa0h accounted for only 4.20% quantified proteins and 3.23% differentially expressed proteins\xa0(DEPs), respectively, much higher percentages of correlations (30.56%-59.11%) were found for other comparative groups (i.e., 18\xa0h vs 14\xa0h, 20\xa0h vs 14\xa0h, and 22\xa0h vs 14\xa0h). According to Spearman correlation tests between transcriptome and proteome (the absolute value of a correlation coefficient between 0.5 and 1 indicates a strong correlation), poor correlations were found for all quantified proteins ( = -\u20090.0355 to 0.0138), DEPs ( = -\u20090.0079 to 0.0233) and the DEPs with opposite expression trends to corresponding differentially expressed genes (DEGs) ( = -\u20090.0478 to 0.0636), whereas stronger correlations were observed in terms of the DEPs with the same expression trends as the correlated DEGs ( = 0.5593 to 0.7080). The results of multiple reaction monitoring (MRM) verification indicate that the iTRAQ results were reliable. After the fermentation arrest, a number of proteins involved in transcription, translation, oxidative phosphorylation and fatty metabolism were down-regulated, some molecular chaperones and proteasome proteins were up-regulated, the ATPase activity significantly decreased, and the total fatty acids gradually accumulated. In addition, the contents of , oleic , C16, C18, C22 and C24 fatty acids increased by 16.77%, 28.49%, 14.14%, 26.88%, 628.57% and 125.29%, respectively.This study confirmed some biochemical and enzymatic alterations provoked by the stress conditions in the specific case of : such as decreases in transcription, translation and oxidative phosphorylation, alterations in cellular fatty composition, and increases in the abundance of molecular chaperones and proteasome proteins. These findings provide potential targets for further metabolic engineering towards improvement of the stress tolerance in .

Keyword: energy

Integration of proteomics and metabolomics data of early and middle season Hass avocados under heat treatment.

Ripening heterogeneity of Hass avocados results in inconsistent quality fruit delivered to the triggered and ready to eat markets. This research aimed to understand the effect of a heat shock (HS) prior to controlled atmosphere (CA) storage on the reduction of ripening heterogeneity. HS prior to CA storage reduces more drastically the ripening heterogeneity in middle season fruit. Via correlation network analysis we show the different metabolomics networks between HS and CA. High throughput proteomics revealed 135 differentially expressed proteins unique to middle season fruit triggered by HS. Further integration of metabolomics and proteomics data revealed that HS reduced the glycolytic throughput and induced protein degradation to deliver for the alternative ripening pathways. l-isoleucine, l-valine, l-aspartic and ubiquitin carboxyl-terminal hydrolase involved in protein degradation were positively correlated to HS samples. Our study provides new insights into the effectiveness of HS in synchronizing ripening of Hass avocados.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

Nutritive Value of Desert Truffles Species of Genera Terfezia and Picoa (Ascomycetes) from Arid and Semiarid Regions of Eastern Turkey.

Terfezia and Picoa species contained 63-94 kcal , 76.93-83.81 g moisture, 0.78-1.52 g ash, 2.19-4.69 g protein, 2.14-9.48 g carbohydrate, 2.54-11.23 g dietary fiber, and 0.96-3.40 g fat per 100 g wet weight. We determined that Terfezia species contain more vitamin E and malondialdehyde than do Picoa species, but the amounts of vitamin A, vitamin C, and β-carotene may vary. Picoa species contain less and stearic acids but more oleic than do Terfezia species. Amino analyses revealed that glutamic and aspartic were the most abundant. We also noted that Picoa species have larger amounts of glucose and fructose than do Terfezia species. In addition, the elements these mushrooms contain can vary, but they are found at nutritious levels and are below toxic levels.

Keyword: energy

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in lipid metabolism during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin metabolism. We further demonstrated the impact of cellular lipid metabolism, and particularly cardiolipin metabolism, during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: energy

Hemin Improves Insulin Sensitivity and Lipid Metabolism in Cultured Hepatocytes and Mice Fed a High-Fat Diet.

Hemin is a breakdown product of hemoglobin. It has been reported that the injection of hemin improves lipid metabolism and insulin sensitivity in various genetic models. However, the effect of hemin supplementation in food on lipid metabolism and insulin sensitivity is still unclear, and whether hemin directly affects cellular insulin sensitivity is yet to be elucidated. Here we show that hemin enhances insulin-induced phosphorylation of insulin receptors, Akt, Gsk3β, FoxO1 and cytoplasmic translocation of FoxO1 in cultured primary hepatocytes under insulin-resistant conditions. Furthermore, hemin diminishes the accumulation of triglyceride and increases in free fatty content in primary hepatocytes induced by palmitate. Oral administration of hemin decreases body weight, intake, blood glucose and triglyceride levels, and improves insulin and glucose tolerance as well as hepatic insulin signaling and hepatic steatosis in male mice fed a high-fat diet. In addition, hemin treatment decreases the mRNA and protein levels of some hepatic genes involved in lipogenic regulation, fatty synthesis and storage, and increases the mRNA level and enzyme activity of CPT1 involved in fatty oxidation. These data demonstrate that hemin can improve lipid metabolism and insulin sensitivity in both cultured hepatocytes and mice fed a high-fat diet, and show the potential beneficial effects of hemin from food on lipid and glucose metabolism.

Keyword: energy

Utilization of milk fatty acids by the suckling Iberian piglets.

A total of 16 pure-bred Iberian (IB) sows, all of them suckling six piglets, were used, eight of them in each of the two consecutive trials (1 and 2). Daily milk yield and composition were determined weekly over a 34-day lactation period. Within each litter, one piglet at birth and four piglets on day 35 of life were slaughtered. Milk intake per piglet tended to be greater in trial 2 (832 v. 893 g/day; P=0.066), but piglets grew at 168±3.3 g/day, irrespective of the trial. In the IB sow milk, the linoleic (LA) : linolenic (LNA) ratio averaged 14.6 and 15.2 in trial 1 and trial 2, respectively. A fivefold increase in piglet body fat content was observed over lactation (P<0.001). Most of this fat (81.4%) was present in the carcass. After 34 days of lactation, whole-body relative content of , palmitoleic, stearic and oleic acids were very close to those in the milk consumed, suggesting direct deposition. Daily deposition of LA derivatives and of LNA and its derivatives was found to be extremely low (<0.02 g, on average). Moreover, some of the arachidonic (ARA) in tissues of the IB piglet at birth disappeared throughout the lactating period. An overall fractional deposition for total fatty acids (FA) was 0.409. Fractional oxidation (disappearance) rates were 0.939 and 0.926 for n-6 and n-3 polyunsaturated FA. The overall rate of disappearance for the major non-essential FA (myristic, , palmitoleic, stearic and oleic acids), estimated as 1-the overall fractional deposition rate, was 0.546. It is concluded that the high degree of FA unsaturation, high oxidation rate of LA and LNA, and poor synthesis of ARA from LA and of docosahexaenoic from LNA found in the suckling piglet might increase the cost of whole-body fat accretion, a contributor to the observed low efficiency of use of milk for growth.

Keyword: energy

Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk fat content (3.60 vs. 3.41%), milk fat yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% fat-corrected milk (47.6 vs. 46.5 kg/d), and -corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk fat yield (1.70 vs. 1.66 kg/d), and 3.5% fat-corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk fat yield, -corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Phthalate exposure and childhood overweight and obesity: Urinary metabolomic evidence.

Metabolomics may unravel global metabolic changes in response to environmental exposures and identify important biological pathways involved in the pathophysiology of childhood obesity. Phthalate has been considered an obesogen and contributing to overweight and obesity in children. The purpose of this study is to evaluate changes in urine metabolites in response to the environmental phthalate exposure among overweight or obese children, and to investigate the metabolic mechanisms involved in the obesogenic effect of phthalate on children at puberty.Within the national Puberty Timing and Health Effects in Chinese Children (PTHEC) study, 69 overweight/obese children and 80 normal weight children were selected into the current study according to their puberty timing and WGOC (The Working Group for obesity in China) references. Urinary concentrations of six phthalate monoesters (MMP, MEP, MnBP, MEHP, MEOHP and MEHHP) were measured using API 2000 electrospray triple quadrupole mass spectrometer (ESIMS/MS). Metabolomic profiling of spot urine was performed using gas chromatography-mass spectrometry. Differentially expressed urinary metabolites associated with phthalate monoesters exposure were examined using orthogonal partial least square-discriminant analysis and multiple linear regression models. In addition, the candidate metabolites were regressed to obesity indices with multiple linear regression models and logistic regression models in all subjects.Compared with normal weight children, higher levels of MnBP were detected in urinary samples of children with overweight and obesity. After adjusting for confounders including chronological age, gender, puberty onset, daily intake and physical activity and socio-economic level, positive association remained between urinary MnBP concentration and childhood overweight/obesity [OR\u202f=\u202f1.586, 95% CI:1.043,2.412]. We observed elevated MnBP concentration was significantly correlated with increased levels of monostearin, 1-monopalmitin, stearic , itaconic , glycerol 3-phosphate, 5-methoxytryptamine, kyotorphin, 1-methylhydantoin, d-alanyl-d-alanine, pyrrole-2-carboxylic , 3,4-Dihydroxyphenylglycol, and butyraldehyde. Meanwhile, increased MnBP concentration was also significantly correlated with decreased levels of lactate, glucose 6-phosphate, d-fructose 6-phosphate, , 4-acetamidobutyric , l-glutamic , n-acetyl-l-phenylalanine, iminodiacetic , hydroxyproline, pipecolinic , l-ornithine, n-acetyl-l-glutamic , guanosine, cytosin, and (s)-mandelic in the normal weight subjects. The observations indicated that MnBP exposure was related to global urine metabolic abnormalities characterized by disrupting arginine and proline metabolism and increasing oxidative stress and fatty reesterification. Among the metabolic markers related to MnBP exposure, 1-methylhydantoin, pyrrole-2-carboxylic and monostearin were found to be positively correlated with obesity indices, while hydroxyproline, l-ornithine, and lactate were negatively associated with overweight/obesity in children.Our results suggested that the disrupted arginine and proline metabolism associated with phthalate exposure might contribute to the development of overweight and obesity in school-age children, providing insights into the pathophysiological changes and molecular mechanisms involved in childhood obesity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Efficacy of bezafibrate on fibroblasts of glutaric acidemia type II patients evaluated using an in vitro probe acylcarnitine assay.

We evaluated the effects of bezafibrate (BEZ) on β-oxidation in fibroblasts obtained from patients with glutaric acidemia type II (GA2) of various clinical severities using an in vitro probe (IVP) assay.Cultured fibroblasts from 12 patients with GA2, including cases of the neonatal-onset type both with and without congenital anomalies (the prenatal- and neonatal-onset forms, respectively), the infantile-onset, and the myopathic forms, were studied. The IVP assay was performed by measuring acylcarnitines (ACs) in the cell culture medium of fibroblasts incubated with for 96h in the presence of 0-800μM BEZ using tandem mass spectrometry.The IVP assay showed that 100μM BEZ markedly reduced the level of palmitoylcarnitine (C16) in the neonatal-onset, infantile-onset, and myopathic forms of GA2, either increasing or maintaining a high level of acetylcarnitine (C2), which serves as an index of production via β-oxidation. In the prenatal-onset form, although a small reduction of C16 was also observed in the presence of 100μM BEZ, the level of C2 remained low. At concentrations higher than 100μM, BEZ further decreased the level of ACs including C16, but a concentration over 400μM decreased the level of C2 in most cases.BEZ at 100μM was effective for all GA2 phenotypes except for the prenatal-onset form, as a reduction of C16 without deterioration of C2 is considered to indicate improvement of β-oxidation. The effects of higher doses BEZ could not be estimated by the IVP assay but might be small or nonexistent.Copyright © 2016 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

Keyword: energy

Antibacterial activity of extracted bioactive molecules of Schinus terebinthifolius ripened fruits against some pathogenic bacteria.

The aim of this work is to identify the chemical constituents and the bioactivity of essential oil (EO), acetone extract (ACE) and n-hexane extract (HexE) of S. terebinthifolius ripened fruits using GC-MS. Total phenolic content and antioxidant activity of extracts were determined using the Folin-Ciocalteu and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assays, respectively. The toxicity against the growth of Acinetobacter baumannii, Bacillus subtilis, Escherichia coli, Micrococcus flavus, Pseudomonas aeruginosa, Sarcina lutea, and Staphylococcus aureus was determined with measuring the inhibition zones (IZs) using the disc diffusion method at the concentrations from 125 to 2000\u202fμg/mL, also, the minimum inhibitory concentrations (MICs) using 96-well micro-plates and ranged from 4 to 2000\u202fμg/mL. The major components in EO were α-pinene (36.9%), and α-phellandrene (32.8%). The major components in ACE were oleic (38.7%), α-phellandrene (13.33%), and δ-cadinene (11.1%), while the major methyl esters of fatty acids detected in HexE were oleic (12.8%), and (10.9%). The EO showed good activity against the growth of Staph. aureus and P. aeruginosa with MIC values of 16\u202fμg/mL and 32\u202fμg/mL, the ACE showed broad activity against the studied bacterial pathogens with MIC values ranged from of 4-128\u202fμg/mL against the studied bacterial isolates, while HexE, however, showed weak antibacterial activity. The IC values of EO, ACE and HexE were 15.11\u202f±\u202f0.99, 118.16\u202f±\u202f1.7 and 324.26\u202f±\u202f2.45\u202fμg/mL, respectively, compared to IC of Tannic (23.83\u202f±\u202f1.9\u202fμg/mL) and butylated hydroxytoluene (BHT, 2.9\u202f±\u202f0.1\u202fμg/mL). Data suggested that the ripened fruits of S. terebinthifolius have potent antioxidant and antibacterial activities.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Superhydrophobic Polypropylene Functionalized with Nanoparticles for Efficient Fast Static and Dynamic Separation of Spilled Oil from Water.

Frequent oil spills not only threaten the ecosystem, but they are also a waste of a valuable source of . There is an urgent need to develop materials that can readily remove spilled oil from water bodies and also have the capacity to collect it for applications. Herein, a superhydrophobic fiber of functionalized polypropylene is engineered with the help of interaction with incorporated copper oxide nanoparticles. The successful development of functionalized polypropylene is confirmed by Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, and -dispersive X-ray spectroscopy. The scanning electron microscopy images reveal that the surface roughness of the polypropylene is enhanced after functionalization. The optimized functionalized polypropylene displays an ultrahydrophobic surface with a water contact angle of 162.42°. The functionalized polyprolyene displays good absorption capacity. It has the capacity to take 30 to 40 times its own weight in oils and nonpolar organic solvents, which makes it useful for small spills. With a flux of 11 204 Lm h, functionalized polypropylene is as an ideal material for the dynamic separation of oil spills from water. It also has excellent selectivity towards oil, water rejection, and oil absorption capacity.© 2019 The Authors. Published by WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: energy

[THE UNESTERIFIED FATTY ACIDS IN BLOOD PLASMA AND INTERCELLULAR MEDIUM: EFFECT OF INSULIN AND ALBUMIN (THE LECTURE)].

The high content of saturated fatty , triglycerides in food, the large amount of lipoproteins of very low density of the same name in blood, obvious insufficient amount of unesterified fatty acids releasing under lipolysis in blood to meet in vivo requirements in biotransforming of ATP are the causes of biological malfunction of homeostasis. As a rule, for every cell in vivo everything is always to be enough. The deficiency of synthesis of ATP by reason of non-optimal substratum for acquirement of ATP by mitochondria is followed by activation also phylogenetically earlier biological function of adaptation, biological reaction of stress. Thus, surplus of unesterified fatty after every food intake forms in vivo biological reaction of "metabolic" stress, deficiency of by reason of realization by mitochondria in vivo non-optimal exogenous substratum- unesterified fatty , deficiency of acyl- and acetyl-KoA and prognostically formation of potentially ineffective alternative of metabolism of fatty acids. The deficiency of unesterified fatty acids in biological reaction of exotrophy after every intake of food compensates biological reaction of stress, activation of releasing of unesterified fatty acids from visceral fatty cells of gland as it physiologically occurs in biological reaction of endotrophy. At that, adrenalin increases lipolysis in visceral fatty cells of gland and physiologically late insulin can\'t inhibit lipolysis in phylogenetically early visceral fatty cells. Increasing of content of unesterifed fatty acids in blood plasma, as it always occurs in vivo, stops absorption of glucose by cells initiating hyperglycemia, hyperinsulinemia, and syndrome of resistance to insulin. The result of such a compensation of biological reaction of exotrophy is biological reaction of endotrophy, condition of "metabolic" stress, depletion of function of β-cells of islets with formation of diabetes mellitus type I, deficiency in vivo of insulin synthesis. The biological role of albumin - transfer of fatty acids in intercellular medium inform of unesterifed fatty acids and prevention of formation of pool of free fatty acids effecting aphysiologically.

Keyword: energy

Microalgae Characterization for Consolidated and New Application in Human Food, Animal Feed and Nutraceuticals.

The exploration of new food sources and natural products is the result of the increase in world population as well as the need for a healthier diet; in this context, microalgae are undoubtedly an interesting solution. With the intent to enhance their value in new commercial applications, this paper aims to characterize microalgae that have already been recognized as safe or authorized as additives for humans and animals (, , , ) as well as those that have not yet been marketed ( and sp.). In this scope, the content of proteins, carbohydrates, lipids, total dietary fiber, humidity, ash, and carotenoids has been measured via standard methods. In addition, individual carotenoids (beta-carotene, astaxanthin, and lutein) as well as individual saturated, monounsaturated, and polyunsaturated fatty acids have been identified and quantified chromatographically. The results confirm the prerogative of some species to produce certain products such as carotenoids, polyunsaturated fatty acids, and proteins, but also show how their cellular content is rich and diverse. green and red phases, and sp., in addition to producing astaxanthin and omega-3, contain about 25⁻33% / proteins on a dry basis. is rich in beta-carotene (3.45% / on a dry basis), is a source of lutein (0.30% / on a dry basis), and the species is a protein-based microalgae (45% / on a dry basis). All, however, can also produce important fatty acids such as , γ-linolenic , and oleic . Considering their varied composition, these microalgae can find applications in multiple sectors. This is true for microalgae already on the market as well as for promising new sources of bioproducts such as and sp.

Keyword: energy

Central composite design parameterization of microalgae/cyanobacteria co-culture pretreatment for enhanced lipid extraction using an external clamp-on ultrasonic transducer.

Lipids extracted from algal biomass could provide an abundant, rapidly growing, high yield feedstock for bio-diesel and other green fuels to supplement current fossil-based sources. Ultrasound pretreatment is a mechanical cell disruption method that has been shown to enhance lipid recovery from algae due to cavitation effects that disrupt algae cell walls. In this study, a locally grown mixture of Chlorella vulgaris/Cyanobacteria leptolyngbya was sonicated in an ultrasonic reactor with a clamp-on transducer prior to solvent lipid extraction. This configuration allows for a non-contact delivery method of ultrasonic with improved operational advantages (no fouling of transducer, continuous operation, and fully scalable design). A central composite design (CCD) was implemented to statistically analyze and evaluate the effect of ultrasonic power (350-750\u202fW) and treatment time (5-30\u202fmin) on lipid yield. Lipid recovery was found to increase with both ultrasonic power and treatment time. Total lipid yields (on dry biomass basis) extracted via the Bligh and Dyer method from Chlorella vulgaris/cyanobacteria co-culture ranged from 8.3% for untreated algae to 16.9% for algae sonicated with 750\u202fW power for 30\u202fmin, which corresponds to more than a doubling of lipid recovery due to ultrasound pretreatment. Increased power and treatment times were found to increase the degree of cell disruption as observed in the SEM and TEM images after ultrasonic pretreatment. Additionally, hexane (1:1 v/v) was evaluated as an alternative to the standard Bligh & Dyer (2:2:1.8 v/v/v chloroform/methanol/cell suspension) lipid extraction solvent system. On average, the Bligh and Dyer method extracted on average over twice the amount of lipids compared to hexane extraction. The lipid profile of the algae extracts indicates high concentrations of lauric (12:0), (16:0), stearic (18:0), oleic (18:1), and linoleic (18:2). This particular configuration of an ultrasonic system proved to be a viable method for the pretreatment of algae for enhanced lipid yields. Future research should focus on identifying alternative extraction solvents and expanding the range of treatment conditions to optimize the ultrasonic power and treatment times for maximum lipid recovery.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

is a novel hypothalamic gene upregulated by a high-fat diet and leptin in mice.

homeostasis is regulated by the hypothalamus but fails when animals are fed a high-fat diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated fat, oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic inflammation.

Keyword: energy

Lipids in human milk.

Fat is the main providing component in human milk and comprising a complex mixture of different lipid species, with quantitative dominance of triglycerides. After elucidating the fatty composition, more recent research has looked at influencing factors and the importance of specific lipids. Here we review quantitative aspects of maternal metabolism which contribute to the milk fatty composition, especially considering essential fatty acids and their long chain polyunsaturated derivatives. In this context studies with stable isotopes have indicated the importance of maternal body pools for mediating the effects of diet on milk composition. Furthermore, the importance of positioning of at the\xa0glycerol backbone of triglycerides is discussed, and the phospholipids of the milk fat globule membrane are described and examples for their potential importance for infant development are presented.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

New compounds inhibiting lipid accumulation from the stems of Sabia parviflora.

Four new compounds, sabianin A (1), sabianin B (2), sabianin C, (3) and sabianin D (4) together with seven known compounds (5-11) were isolated from the stems of Sabia parviflora. The structures of these compounds were elucidated by extensive spectroscopic analysis and by comparing their NMR data with those of related compounds. Their absolute configurations were determined by comparing the calculated and experimental electronic circular dichroism spectra. The lipid-lowering effect of all the compounds was evaluated using a cell model of steatosis induced by oleic and . Compounds 3, 6, and 10 showed statistically significant lipid-lowering effect.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

Metabolic changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Effect of flue gas CO on the growth, carbohydrate and fatty composition of a green microalga Scenedesmus obliquus for biofuel production.

Effect of various flue gas CO concentrations (5%, 10% and 14.1%) on growth rate and biochemical properties of a green microalga Scenedesmus obliquus was investigated. S. obliquus showed the highest biomass production and growth rate (0.36\u2005g\u2005L and μ\u2009=\u20091.00\u2005day), total inorganic carbon removal (35.8\u2005mg\u2005L), lipid productivity (9.9\u2005mg\u2005L\u2005day) and carbohydrate productivity (10.3\u2005mg\u2005L\u2005day) with 14.1% CO after 8 days of cultivation. Fatty methyl ester analysis revealed that the and oleic contents were increased up to 5% and 7% with 14.1% CO, respectively. Application of flue gas CO enhanced the growth along with lipid and carbohydrate productivity of S. obliquus, which can be exploited for reducing the CO concentration.

Keyword: energy

Effect of on the mitigation of milk fat depression syndrome caused by trans-10, cis-12-conjugated linoleic in grazing dairy cows.

The objective of the study was to evaluate the effect of adding protected (PA) to the ration of grazing dairy cows supplemented with protected conjugated linoleic (CLA) on milk production, chemical composition and fat profile. Six cows were used, 3/4 American Swiss\xa0×\xa0Zebu, under a rotational grazing system in a mixed sward with Cynodon plectostachyus, Brachiaria decumbens and Brachiaria brizantha. Furthermore, each cow received daily 4\xa0kg concentrates and 8\xa0kg sorghum silage, which made up the basal diet. The cows were distributed into three two-cow groups. Three treatments were randomly assigned to the groups, using a cross design: (1) control (basal diet), (2) basal diet\xa0+\xa0CLA (50\xa0g/d) and (3) basal diet\xa0+\xa0CLA (50\xa0g/d)\xa0+\xa0PA (412\xa0g/d). The following variables were evaluated: forage intake, milk production, protein, fat and lactose concentration in milk, and milk fatty (FA) profile. There were no differences in forage intake between treatments; however, there were differences in milk production, protein, fat and lactose yield and fat concentration, which increased significantly in group CLA\xa0+\xa0PA when compared with group CLA. The concentration of FA synthesised de novo was lower when PA was included in the diet. Adding PA to the diet of grazing cows mitigates the milk fat decline caused by including trans-10, cis-12 CLA in the diet.

Keyword: energy

Impact of Maternal Diet on Human Milk Composition Among Lactating Women in Latvia.

: Many studies indicate that the maternal diet is an important factor affecting human milk composition. Human milk composition among lactating women in Latvia, as well as the maternal diet during lactation, has not been sufficiently studied. The aim of this research was to assess dietary habits and macronutrient intake among lactating women in Latvia and to examine the effect of diet on human milk composition. : Research was conducted between November 2016 and December 2017. Mature human milk samples ( = 61) along with a 72h food diary, a food frequency questionnaire (FFQ), and a questionnaire about maternal and infant characteristics were obtained from voluntary women who were recruited via an invitation published in a social media member group for nursing mothers. Fat content in human milk was determined by LVS ISO 2446:2008, protein content was determined by LVS EN ISO 8968-1:2014, lactose was determined by ISO 22662:2007, and the fatty profile was analyzed using gas chromatography. Dietary data were evaluated using the Finnish food composition database Fineli, release 19 (3 March 2018). : Median values for fat, protein, and lactose in mature human milk were 4.40%, 1.08%, and 6.52%, respectively. Predominant fatty acids in human milk were oleic (C18:1 n9), (C16:0), and linoleic (C18:2 n6) at 34.60%, 24.00%, and 11.00% of total fatty acids, respectively. The elaidic (C18:1 n9) level was <0.10% in all human milk samples. Significant, positive associations ( < 0.05) were found between maternal dietary intake of linoleic, α-linolenic, docosahexaenoic, total -monounsaturated, total -polyunsaturated, and total n-6 and n-3 polyunsaturated fatty acids, the ratio of n-6/n-3, and the level of these fatty acids in human milk. Total and carbohydrate intake among participants were lower, but total fat, saturated fat, and sugar intake were higher than recommended. Protein, linoleic , and α-linolenic intake were adequate, but docosahexaenoic intake was noticeably lower than recommended. Women should be supported with information regarding their nutritional needs during lactation and the possible impact of diet on human milk composition. : Macronutrient (fat, protein, and lactose) content in human milk is not affected by maternal diet. Conversely, the human milk fatty profile is affected by the immediate diet consumed by the mother. Habitual dietary habits can also impact the fatty profile of human milk.

Keyword: energy

Intake of individual saturated fatty acids and risk of coronary heart disease in US men and women: two prospective longitudinal cohort studies.

\xa0To investigate the association between long term intake of individual saturated fatty acids (SFAs) and the risk of coronary heart disease, in two large cohort studies.\xa0Prospective, longitudinal cohort study.\xa0Health professionals in the United States.\xa073\u2009147 women in the Nurses\' Health Study (1984-2012) and 42\u2009635 men in the Health Professionals Follow-up Study (1986-2010), who were free of major chronic diseases at baseline.\xa0Incidence of coronary heart disease (n=7035) was self-reported, and related deaths were identified by searching National Death Index or through report of next of kin or postal authority. Cases were confirmed by medical records review.\xa0Mean intake of SFAs accounted for 9.0-11.3% intake over time, and was mainly composed of lauric (12:0), myristic (14:0), (16:0), and stearic (18:0; 8.8-10.7% ). Intake of 12:0, 14:0, 16:0 and 18:0 were highly correlated, with Spearman correlation coefficients between 0.38 and 0.93 (all P<0.001). Comparing the highest to the lowest groups of individual SFA intakes, hazard ratios of coronary heart disease were 1.07 (95% confidence interval 0.99 to 1.15; P=0.05) for 12:0, 1.13 (1.05 to 1.22; P<0.001) for 14:0, 1.18 (1.09 to 1.27; P<0.001) for 16:0, 1.18 (1.09 to 1.28; P<0.001) for 18:0, and 1.18 (1.09 to 1.28; P<0.001) for all four SFAs combined (12:0-18:0), after multivariate adjustment of lifestyle factors and total intake. Hazard ratios of coronary heart disease for isocaloric replacement of 1% from 12:0-18:0 were 0.92 (95% confidence interval 0.89 to 0.96; P<0.001) for polyunsaturated fat, 0.95 (0.90 to 1.01; P=0.08) for monounsaturated fat, 0.94 (0.91 to 0.97; P<0.001) for whole grain carbohydrates, and 0.93 (0.89 to 0.97; P=0.001) for plant proteins. For individual SFAs, the lowest risk of coronary heart disease was observed when the most abundant SFA, 16:0, was replaced. Hazard ratios of coronary heart disease for replacing 1% from 16:0 were 0.88 (95% confidence interval 0.81 to 0.96; P=0.002) for polyunsaturated fat, 0.92 (0.83 to 1.02; P=0.10) for monounsaturated fat, 0.90 (0.83 to 0.97; P=0.01) for whole grain carbohydrates, and 0.89 (0.82 to 0.97; P=0.01) for plant proteins.\xa0Higher dietary intakes of major SFAs are associated with an increased risk of coronary heart disease. Owing to similar associations and high correlations among individual SFAs, dietary recommendations for the prevention of coronary heart disease should continue to focus on replacing total saturated fat with more healthy sources of .Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Keyword: energy

Attenuation of Free Fatty -Induced Muscle Insulin Resistance by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 diabetes mellitus. Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties.

Keyword: energy

Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty composition.

A profound analysis of A.\xa0tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty profile. Plants exposed to 125-250\xa0mm NaCl showed higher photochemical light harvesting efficiencies and lower dissipation rates. under higher NaCl exposure, there is evident damage of the oxygen evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the fluxes entering the PS II, in spite of the high amounts of dissipated and the loss of PS II antennae connectivity. dissipation was mainly achieved through the auroxanthin pathway. Total fatty content displayed a similar trend, being also higher under 125-250\xa0mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic and allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250\xa0mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.© 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Keyword: energy

Environmental temperature alters the overall digestive energetics and differentially affects dietary protein and lipid use in a lizard.

Processing food (e.g. ingestion, digestion, assimilation) requires referred to as specific dynamic action (SDA) and is at least partially fuelled by oxidation of the nutrients (e.g. proteins and lipids) within the recently ingested meal. In ectotherms, environmental temperature can affect the magnitude and/or duration of the SDA, but is likely to also alter the mixture of nutrients that are oxidized to cover these costs. Here, we examined metabolic rate, gut passage time, assimilation efficiency and fuel use in the lizard digesting cricket meals at three ecologically relevant temperatures (20, 25 and 32°C). Crickets were isotopically enriched with C-leucine or C- tracers to distinguish between protein and lipid oxidation, respectively. Our results show that higher temperatures increased the magnitude of the SDA peak (by 318% between 32 and 20°C) and gut passage rate (63%), and decreased the duration of the SDA response (by 20% for males and 48% for females). Peak rate of dietary protein oxidation occurred sooner than peak lipid oxidation at all temperatures (70, 60 and 31\u2005h earlier for 20, 25 and 32°C, respectively). Assimilation efficiency of proteins, but not lipids, was positively related to temperature. Interestingly, the SDA response exhibited a notable circadian rhythm. These results show that temperature has a pronounced effect on digestive energetics in , and that this effect differs between nutrient classes. Variation in environmental temperatures may thus alter the budget and nutrient reserves of these animals.© 2019. Published by The Company of Biologists Ltd.

Keyword: energy

Feeding of palm oil fatty acids or rapeseed oil throughout lactation: Effects on status, body composition, and milk production in Norwegian dairy goats.

The objective of this experiment was to examine how supplements of rapeseed oil or palm oil fatty acids would affect milk production and composition, body lipid stores, and balance in 30 multiparous goats of Norwegian dairy goat breed. The experiment lasted 230 d, with 1 to 120 d in milk (DIM) for indoor feeding (P1), 120 to 200 DIM for mountain grazing (P2), and 200 to 230 DIM for indoor feeding (P3). Grass silage was fed according to appetite during indoor feeding periods. After an adjustment period (1-60 DIM) when the control diet was given to the goats, the animals were subdivided into 3 groups of 10 goats. Treatments (60-230 DIM) were (1) basal concentrate (control; no added fat); (2) control concentrate with 8% (added on air-dry basis) hydrogenated palm oil enriched with (POFA); and (3) control concentrate with 8% (added on air-dry basis) rapeseed oil (RSO). Individual balances based on intake and milk production were estimated on 10, 30, 60, 90, 120, 200, and 230 DIM. At the same times, body weight (BW), body condition score (BCS), body mass index, and body tissue stores using computed tomography were monitored. Silage intake was depressed by POFA throughout the experimental period. Reduced BW and body mass index were observed in the POFA and RSO groups, whereas no effect on BCS or body composition was observed throughout lactation. Generally, a minor decrease in BW was observed from 10 to 120 DIM (only 0.6 kg on average) and the total amount of body lipid was reduced by 4.4 kg. During the mountain grazing period, a further reduction in body lipid stores (2.7 kg) was observed, and BW was reduced by 3.9 kg in the same period. The goats mobilized, on average, 72% of their fat reserves during the first 200 DIM. In this period, dietary fat supplementation did not reduce the mobilization of adipose tissue but resulted in greater milk fat yield (2 kg more, on average, compared with the control group). Milk yield was not affected by POFA or RSO supplementation. Milk fat content was higher in the POFA group than in the control and RSO groups. Milk protein and lactose contents were not affected by lipid supplements. In late lactation, a rapid accumulation of fat deposits followed the intense mobilization during the grazing period. Dietary lipid supplements had no effect on milk fat yield at this stage. Milk production depends heavily on the ability to mobilize body lipid stores, and neither POFA nor RSO supplements at rates used in our study affected this mobilization.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Dihydrosterculic from cottonseed oil suppresses desaturase activity and improves liver metabolomic profiles of high-fat-fed mice.

Polyunsaturated fatty (PUFA)-rich diets are thought to provide beneficial effects toward metabolic health in part through their bioactive properties. We hypothesized that increasing PUFA intake in mice would increase peroxisome proliferator activated receptor delta (PPARδ) expression and activity, and we sought to examine the effect of different PUFA-enriched oils on muscle PPARδ expression. One of the oils we tested was cottonseed oil (CSO) which is primarily linoleic (53%) and (24%). Mice fed a CSO-enriched diet (50% from fat) displayed no change in muscle PPARδ expression; however, in the liver, it was consistently elevated along with its transcriptional coactivator Pgc-1. Male mice were fed chow or CSO-, saturated fat (SFA)-, or linoleic (18:2)-enriched diets that were matched for macronutrient content for 4 weeks. There were no differences in food intake, body weight, fasting glucose, glucose tolerance, or expenditure between chow- and CSO-fed mice, whereas SFA-fed mice had increased fat mass and 18:2-fed mice were less glucose tolerant. Metabolomic analyses revealed that the livers of CSO-fed mice closely matched those of chow-fed but significantly differed from SFA- and 18:2-enriched groups. Fatty composition of the diets and livers revealed an impairment in desaturase activity and the presence of dihydrosterculic (DHSA) in the CSO-fed mice. The effect of DHSA on PPARδ and stearoyl-CoA desaturase-1 expression mimicked that of the CSO-fed mice. Taken together, these data suggest that DHSA from CSO may be an effective means to increase PPARδ expression with concomitant suppression of liver stearoyl-CoA desaturase-1 activity.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Multi-omic profiles of hepatic metabolism in TPN-fed preterm pigs administered new generation lipid emulsions.

We aimed to characterize the lipidomic, metabolomic, and transcriptomic profiles in preterm piglets administered enteral (ENT) formula or three parenteral lipid emulsions [parenteral nutrition (PN)], Intralipid (IL), Omegaven (OV), or SMOFlipid (SL), for 14 days. Piglets in all parenteral lipid groups showed differential organ growth versus ENT piglets; whole body growth rate was lowest in IL piglets, yet there were no differences in either expenditure or (13)C-palmitate oxidation. Plasma homeostatic model assessment of insulin resistance demonstrated insulin resistance in IL, but not OV or SL, compared with ENT. The fatty and acyl-CoA content of the liver, muscle, brain, and plasma fatty acids reflected the composition of the dietary lipids administered. Free carnitine and acylcarnitine (ACT) levels were markedly reduced in the PN groups compared with ENT piglets. Genes associated with oxidative stress and inflammation were increased, whereas those associated with alternative pathways of fatty oxidation were decreased in all PN groups. Our results show that new generation lipid emulsions directly enrich tissue fatty acids, especially in the brain, and lead to improved growth and insulin sensitivity compared with a soybean lipid emulsion. In all total PN groups, carnitine levels are limiting to the formation of ACTs and gene expression reflects the stress of excess lipid on liver function.

Keyword: energy

Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced obesity.

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti-obesity drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased body and liver weights, insulin, leptin, triglyceride, cholesterol and free fatty plasma levels in a mouse model of diet-induced obesity. Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in obesity and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

Keyword: energy

Dynamics of Individual Fatty Acids in Muscle Fat Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, metabolic rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C to C diet shift experiment and compound-specific δC analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and lipid droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic (LA; 18:2n6) and (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic (20:4n6) and docosahexaenoic (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular lipid droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in metabolism. The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects metabolism of birds during migration.

Keyword: energy

High density lipoprotein (HDL) reverses induced metabolism imbalance by switching CD36 and GLUT4 signaling pathways in cardiomyocyte.

In our previous study (PA) induced lipotoxicity and switches metabolism from CD36 to GLUT4 in H9c2 cells. Low level of high density lipoprotein (HDL) is an independent risk factor for cardiac hypertrophy. Therefore, we in the present study investigated whether HDL can reverse PA induced lipotoxicity in H9c2 cardiomyoblast cells. In this study, we treated H9c2 cells with PA to create a hyperlipidemia model in vitro and analyzed for CD36 and GLUT4 metabolic pathway proteins. CD36 metabolic pathway proteins (phospho-AMPK, SIRT1, PGC1α, PPARα, CPT1β, and CD36) were decreased by high PA (150 and 200\u2009μg/μl) concentration. Interestingly, expression of GLUT4 metabolic pathway proteins (p-PI3K and pAKT) were increased at low concentration (50\u2009μg/μl) and decreased at high PA concentration. Whereas, phospho-PKCζ, GLUT4 and PDH proteins expression was increased in a dose dependent manner. PA treated H9c2 cells were treated with HDL and analyzed for cell viability. Results showed that HDL treatment induced cell proliferation efficiency in PA treated cells. In addition, HDL reversed the metabolic effects of PA: CD36 translocation was increased and reduced GLUT4 translocation, but HDL treatment significantly increased CD36 metabolic pathway proteins and reduced GLUT4 pathway proteins. Rat neonatal cardiomyocytes showed similar results. In conclusion, HDL reversed palmatic -induced lipotoxicity and metabolism imbalance in H9c2 cardiomyoblast cells and in neonatal rat cardiomyocyte cells.© 2017 Wiley Periodicals, Inc.

Keyword: energy

A new approach of microalgal biomass pretreatment using deep eutectic solvents for enhanced lipid recovery for biodiesel production.

The biomass of Chlorella sp. was pretreated with three different aqueous deep eutectic solvents (aDESs), i.e. aqueous choline chloride-oxalic (aCh-O), aqueous choline chloride-ethylene glycol (aCh-EG) and aqueous urea-acetamide (aU-A). The effect of aDESs pretreatment of microalgae biomass was evaluated in terms of lipid recovery rate, total carbohydrate content, fatty composition, and thermal chemical behavior of biomass. Results indicated that, lipid recovery rate was increased from 52.03% of untreated biomass to 80.90%, 66.92%, and 75.26% of the biomass treated by aCh-O, aCh-EG and aU-A, respectively. However, there were no major changes observed in fatty profiles of both untreated and treated biomass, specifically , palmitoleic and stearic under various pretreatments. Furthermore, characterizations of untreated and treated biomass were carried out using Fourier transform infrared (FTIR), thermogravimetry analysis (TGA) and scanning electron microscope (SEM) to understand the enhanced lipids recovery.Copyright © 2016. Published by Elsevier Ltd.

Keyword: energy

1\'-Acetoxychavicol acetate ameliorates age-related spatial memory deterioration by increasing serum ketone body production as a complementary source for neuronal cells.

1\'-Acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Alpinia galangal. Here, we examined the effect of ACA on learning and memory in senescence-accelerated mice prone 8 (SAMP8). In mice that were fed a control diet containing 0.02% ACA for 25 weeks, the learning ability in the Morris water maze test was significantly enhanced in comparison with mice that were fed the control diet\xa0alone. In the Y-maze test, SAMP8 mice showed decreased spontaneous alterations in comparison with senescence-accelerated resistant/1 (SAMR1) mice, a homologous control, which was improved by ACA pretreatment. Serum metabolite profiles were obtained by GC-MS analysis, and each metabolic profile was plotted on a 3D score plot. Based upon the diagram, it can be seen that the distribution areas for the three groups were completely separate. Furthermore, the contents of β-hydroxybutyric and in the serum of SAMP8-ACA mice were higher than those of SAMP8-control mice and SAMR1-control mice. We also found that SAMR1 mice did not show histological abnormalities, whereas histological damage in the CA1 region of the hippocampus in SAMP8-control mice was observed. However, SAMP8-ACA mice were observed in a similar manner as SAMR1 mice. These findings confirm that ACA increases the serum concentrations of β-hydroxybutyric and levels and thus these fuels might contribute to the maintenance of the cognitive performance of SAMP8 mice.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: energy

Comparison of diets enriched in stearic, oleic, and acids on inflammation, immune response, cardiometabolic risk factors, and fecal bile concentrations in mildly hypercholesterolemic postmenopausal women-randomized crossover trial.

Direct comparisons between SFAs varying in chain length, specifically (16:0) and stearic (18:0), relative to the latter\'s metabolic product, oleic (18:1), on cardiometabolic risk factors are limited.The aim of this study was to determine the relative comparability of diets enriched in , stearic , and oleic on inflammation and coagulation markers, T lymphocyte proliferation/ex-vivo cytokine secretion, plasma cardiometabolic risk factors, and fecal bile concentrations.Hypercholesterolemic postmenopausal women (n\xa0=\xa020, mean\xa0±\xa0SD age 64\xa0±\xa07 y, BMI 26.4\xa0±\xa03.4 kg/m2, LDL cholesterol\xa0≥\xa02.8 mmol/L) were provided with each of 3 diets [55% (%E) carbohydrate, 15%E protein, 30%E fat, with ∼50% fat contributed by , stearic , or oleic in each diet; 5 wk/diet phase] using a randomized crossover design with 2-wk washouts between phases. Outcome measures were assessed at the end of each phase.Fasting LDL-cholesterol and non-HDL-cholesterol concentrations were lower after the stearic and oleic diets than the diet (all P\xa0<\xa00.01). Fasting HDL-cholesterol concentrations were lower after the stearic diet than the and oleic diets (P\xa0<\xa00.01). The stearic diet resulted in lower lithocholic (P\xa0=\xa00.01) and total secondary bile (SBA) concentrations (P\xa0=\xa00.04) than the oleic diet. All other outcome measures were similar between diets. Lithocholic concentrations were positively correlated with fasting LDL-cholesterol concentrations (r\xa0=\xa00.33; P\xa0=\xa00.011). Total SBA, lithocholic , and deoxycholic concentrations were negatively correlated with fasting HDL cholesterol (r\xa0=\xa0-0.51 to -0.44; P\xa0<\xa00.01) concentrations and positively correlated with LDL cholesterol:HDL cholesterol (r =\xa00.37-0.54; P\xa0<\xa00.01) ratios.Dietary stearic and oleic had similar effects on fasting LDL-cholesterol and non-HDL-cholesterol concentrations and more favorable ones than . Unlike oleic , the hypocholesterolemic effect of stearic may be mediated by inhibition of intestinal hydrophobic SBA synthesis. These findings add to the data suggesting there should be a reassessment of current SFA dietary guidance and Nutrient Facts panel labeling.This trial was registered at clinicaltrials.gov as .Copyright © American Society for Nutrition 2019.

Keyword: energy

[CHANGES IN THE METABOLISM IN THE MYOCARDIUM OF RATS WITH ARTERIAL HYPERTENSION].

In the myocardium of the rats with arterial hypertension marked increase in the amount of unsaturated fatty acids and polyunsaturated fatty acids. Reducing the concentration of and increased levels of arachidonic is considered as one of the factors that lead to the development of deficit and oxidative stress. In rats, with hypertension myocardial lactate concentration increases in the cytoplasmic fraction and reducing the amount of ATP. The level in the cytoplasmic and mitochondrial fractions above benchmarks, indicating about the change of antioxidant systems of the body In the cytoplasm and mitochondria of cardiomyocytes of the rats with arterial hypertension marked decrease in the activity of antioxidant enzymes: NO-synthase, catalase, glutathione reductase, which causes metabolic changes of the myocardium.

Keyword: energy

Suitability of lemon balm (Melissa officinalis L.) extract rich in rosmarinic as a potential enhancer of functional properties in cupcakes.

Melissa officinalis (lemon balm) and its extracts have been frequently reported as possessing bioactive properties, offering the potential for use in development/enrichment of food products with additional functional capabilities, providing health benefits to consumers. The antioxidant, antibacterial and antifungal activity of lemon balm extract, as well as its potential hepatotoxicity were thoroughly evaluated. The extracts were then incorporated into cupcakes and their preserving effect, chemical composition, colour parameters and antioxidant activity were compared with those provided by potassium sorbate. In general, the variables with the largest differences among different storage times were level, sucrose, glucose, (C6:0) and oleic (C18:1n9). On the other hand, L (top), a (top), b (top), pH, capric (C10:0) and lauric (C12:0) showed the greatest variation according to cupcake formulation. The results observed indicate that the lemon balm extract rich in rosmarinic can provide advantageous functional properties to bakery products.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Growth, stool consistency and bone mineral content in healthy term infants fed sn-2-palmitate-enriched starter infant formula: A randomized, double-blind, multicentre clinical trial.

Palmitate in breast milk is predominantly located in the triacylglycerol sn-2 position, while infant formulae contain palmitate predominantly in the sn-1 and sn-3 positions. During digestion, palmitate in the sn-1 and sn-3 positions is hydrolyzed to free that can subsequently complex with calcium to form insoluble soaps; this may partially explain why formula-fed infants have harder stools than breast-fed infants.This large (n\xa0=\xa0488) randomized, double-blind, multicentre trial investigated whether increasing the sn-2 palmitate content of infant formula improves stool consistency and bone mineral content (measured by dual- x-ray absorptiometry), without affecting growth or health. From ∼1 week to 4 months of age, infants were exclusively fed one of three formulae: i) control formula (CF; 16% of total palmitate at sn-2; n\xa0=\xa0162), (ii) experimental formula 1 (EF1; 43% of total palmitate at sn-2; n\xa0=\xa0166) or (iii) experimental formula 2 (EF2; 51% of total palmitate at sn-2; n\xa0=\xa0160).Intention-to-treat analysis showed softer stools in both EF groups (vs. CF) at ages 2 weeks and 1 and 2 months (p\xa0≤\xa00.01), but not 3 and 4 months. At 4 months, all groups had similar growth outcomes while bone mineral content was significantly higher in EF1 (p\xa0=\xa00.0012) and EF2 (p\xa0=\xa00.0002) compared with CF. Comparison of reported adverse events up to 12 months revealed no differences among groups. All 3 infant formulae exhibited equally good digestive tolerance.Formulae enriched in sn-2 palmitate fed in early infancy are safe, improve stool consistency (from 2 weeks to 2 months) and increase bone mineral content (at 4 months).Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Keyword: energy

Encapsulation of soybean meal with fats enriched in and stearic acids: effects on rumen-undegraded protein and in vitro intestinal digestibility.

Fat coating of soybean meal (SBM) can reduce its protein degradability in the rumen, but the encapsulation of SBM with (PA) and stearic acids (SA) has not yet been investigated, despite both fatty acids are common sources in dairy cow diets. This study aimed to evaluate the effects of applying a novel method, using either 400 or 500\xa0g fat/kg (treatments FL40 and FL50, respectively), which was enriched in PA and SA at different ratios (100:0, 75:25, 50:50, 25:75 and 0:100), on physical and chemical characteristics, ruminal degradability, solubility and in vitro intestinal protein digestibility (IVIPD) of the obtained products. Encapsulation of SBM in fat resulted in greater mean particle size and lower bulk density and protein solubility than unprotected SBM (USBM). Treatment FL50 resulted in increased (p <\xa00.01) rumen-undegraded protein (RUP) compared to USBM. There were no differences in RUP of SBM when different PA: SA ratios were used. The mean RUP content of treatments FL40 and FL50 (306 and 349\xa0g/kg, respectively) was greater compared to USBM (262\xa0g/kg, p <\xa00.05), but lower than that for a standard heat-treated SBM (431\xa0g/kg). Values of IVIPD did not differ among SBM, heat-treated SBM and FL40 and FL50 samples, all being greater than 97.8%. In conclusion, encapsulation of SBM with fats enriched in PA and SA proved to be effective in reducing protein solubility and increasing RUP without depressing protein digestibility in the intestine. For validation of the method, in vivo research to investigate the effects of these products on the production of dairy cows is warranted.

Keyword: energy

Lipids characterization of ultrasound and microwave processed germinated sorghum.

Cereal crops and oilseeds provide diverse pool of fatty acids with characteristic properties. Sorghum (Sorghum bicolor (L.) Moench) provides the staple food with serving as main source of and protein. Germination of sorghum generally increases the nutritive value of seeds and the effects of germination on lipids composition of seeds vary greatly with processing conditions. Therefore, the current study was conducted to compare the effect of emerging processing techniques such as ultrasound (US) and microwave (MW) on fatty acids composition and oil yield of sorghum seeds before and after germination.Initially sorghum grains were soaked with 5% NaOCl (sodium hypochlorite) for surface sterilization. Afterwards, grains were soaked in excess water for 22\xa0h at room temperature and were divided into four portions. The first portion (100\xa0g grains) was subjected to germination without applying any microwave and ultrasonic treatment (T). Second portion was further divided into four groups (T, T, T, T) (100\xa0g of each group) and grains were subjected to ultrasonic treatments using two different ultrasonic intensities (US: 40%; US: 60%) within range of 0-100% and with two different time durations (t: 5\xa0min; t: 10\xa0min) at constant temperature. Third portion was also divided into four groups (T, T, T, T) (100\xa0g of each group) and exposed to microwave treatments at two different power levels (MW: 450 watt; MW: 700 watt) within the range of 100-900\xa0W for two different time durations (t: 15\xa0s; t: 30s). Similarly, fourth portion was divided into four groups (T, T, T, T) (100\xa0g of each group). Each group was exposed to both MW (MW, MW) (100-900 watt power) & US (US, US) (0-100% intensity) treatments at two different time levels (t, t). Then, germination was carried out and pre-treated raw and pre-treated germinated sorghum grains were analyzed for total oil yield, fatty composition and unsaturated fatty acids (Un-SFA)/saturated fatty acids (SFA) ratio by gas chromatography.The results revealed that oil yield in sorghum before and after germination ranged from 6.55 to 7.84% and 6.28 to 7.57%, respectively. All the microwave and ultrasound processed samples showed significant difference in oil yield than the raw sorghum grains. The highest tested yield was 7.84\xa0±\xa00.31% when combination of microwave power (700\xa0W) and ultrasound intensity (60%) was applied for 30s and 10\xa0min, respectively. The results further demonstrate that the raw sorghum contained (13.73\xa0±\xa00.10%), palmitoleic (0.43\xa0±\xa00.02%), stearic (1.07\xa0±\xa00.04%), oleic (37.15\xa0±\xa00.10%), linoleic (43.33\xa0±\xa00.21%), linolenic (1.55\xa0±\xa00.04%), arachidic (0.13\xa0±\xa00.01%) and eicosenoic (0.37\xa0±\xa00.02%), respectively. The highest fatty percentage for , stearic and arachidic was 13.75\xa0±\xa00.07%, 1.11\xa0±\xa00.09% and 0.15\xa0±\xa00.03% at 60% US intensity for 10\xa0min (T), respectively. Maximum amount observed was 1.60\xa0±\xa00.09% of linolenic while amount of eicosenoic decreased from 0.37\xa0±\xa00.02% to 0.31\xa0±\xa00.01% after processing. In case of applying combination of microwave and sonication treatments, the change in eicosenoic increased from 0.35\xa0±\xa00.02% to 0.40\xa0±\xa00.04% while there was no significant change in other fatty acids. The ungerminated sorghum oil possessed 14.93-15.05% and 82.83-83.12% of SFA and Un-SFA, respectively. After germination, percentage of saturated fatty acids increased (16.4-16.55%) while decreased for unsaturated fatty acids (80.13-80.56%) were noted.The results of the present study conclude that the yield of oil from sorghum grains increased by emerging processing. Fatty analysis of sorghum oil suggested that pre-treatment strategies will not affect the quality of the oil with respect to essential fatty acids content. Overall, the composition of saturated fatty in germinated grain is improved than ungerminated grains after processing.

Keyword: energy

Effects of high-fat diet and AMP-activated protein kinase modulation on the regulation of whole-body lipid metabolism.

Metabolic flexibility, the capacity to adapt to fuel availability for production, is crucial for maintaining whole-body homeostasis. An inability to adequately promote FA utilization is associated with lipid accumulation in peripheral tissues and contributes to the development of insulin resistance. In vivo assays to quantify whole-body lipid oxidation in mouse models of insulin resistance are lacking. We describe a method for assessing whole-body FA oxidation in vivo, as well as tissue-specific lipid uptake in conscious mice. The method relies on intravenous administration of [9,10-H(N)] combined with a non-β-oxidizable palmitate analog, [1-C]2-bromopalmitic . Pretreatment with etomoxir, a CPT1 inhibitor that prevents the shuttling of FAs into mitochondria, markedly reduced the appearance of the β-oxidation product HO in circulation and reduced lipid uptake by oxidative tissues including heart and soleus muscle. Whole-body fatty oxidation was unaltered between chow- or high-fat-fed WT and transgenic mice expressing a mutant form of the AMPK γ3 subunit (AMPKγ3) in skeletal muscle. High-fat feeding increased lipid oxidation in WT and AMPKγ3 transgenic mice. In conclusion, this technique allows for the assessment of the effect of pharmaceutical agents, as well as gene mutations, on whole-body FA oxidation in mice.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: energy

Integrating Extracellular Flux Measurements and Genome-Scale Modeling Reveals Differences between Brown and White Adipocytes.

White adipocytes are specialized for storage, whereas brown adipocytes are specialized for expenditure. Explicating this difference can help identify therapeutic targets for obesity. A common tool to assess metabolic differences between such cells is the Seahorse Extracellular Flux (XF) Analyzer, which measures oxygen consumption and media acidification in the presence of different substrates and perturbagens. Here, we integrate the Analyzer\'s metabolic profile from human white and brown adipocytes with a genome-scale metabolic model to predict flux differences across the metabolic map. Predictions matched experimental data for the metabolite 4-aminobutyrate, the protein ABAT, and the fluxes for glucose, glutamine, and palmitate. We also uncovered a difference in how adipocytes dispose of nitrogenous waste, with brown adipocytes secreting less ammonia and more urea than white adipocytes. Thus, the method and software we developed allow for broader metabolic phenotyping and provide a distinct approach to uncovering metabolic differences.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Role of fatty composites in the toxicity of titanium dioxide nanoparticles used in cosmetic products.

It has been recognized that the use of nanoparticles (NPs) in the cosmetic industry results in products with better efficacy and functionality. However, recent advances in molecular toxicology have revealed that NP exposure can promote cytotoxicity and oxidative damage, which has raised health concerns in the use of NPs in personal care products. Nevertheless, the mechanistic basis for the toxicity and safety of cosmetic NPs is poorly understood. The goal of the study was to determine the cytotoxicity and intracellular distribution of titanium dioxide (TiO2) NPs containing fatty composites (palmitoleic , , stearic and oleic ) commonly used in cosmetic products. Two types of cells, human fibroblast skin cells and adenocarcinoma lung cells, were exposed to either bare TiO2 NPs or TiO2 NPs mixed with fatty acids for up to 48 hr. NMR analysis confirmed that the fatty composites remained in the NPs after wash. The cytotoxicity of TiO2 NPs was determined by cell viability measurement using quantitative confocal microscopy, and the localization of two different forms of TiO2 NPs were assessed using electron spectroscopic imaging with transmission electron microscopy. TiO2 NPs containing fatty acids posed significantly reduced cytotoxicity (80-88% decreases) than bare NPs in both cell types. Furthermore, there was less intracellular penetration of the NPs containing fatty composites compared with bare NPs. These results provide important insights into the role of fatty acids in protecting the cells from possible toxicity caused by NPs used in the production of cosmetic products.

Keyword: energy

Stabilizing mixed fatty and phthalate ester monolayer on artificial seawater.

Phthalate esters which are widely used as industrial chemicals have become widespread contaminants in the marine environment. However, little information is available on the interfacial behavior of phthalate esters in the seawater, where contaminants generally occur at elevated concentrations and have the potential to transfer into the atmosphere through wave breaking on sea surface. We used artificial seawater coated with fatty acids to simulate sea surface microlayer in a Langmuir trough. The interactions of saturated fatty acids (stearic (SA) and (PA)) with one of the most abundant phthalate esters (di-(2-ethylhexyl) phthalate (DEHP)), were investigated under artificial seawater and pure water conditions. Pure DEHP monolayer was not stable, while more stable mixed monolayers were formed by SA and DEHP on the artificial seawater at relatively low surface pressure. Sea salts in the subphase can lower the excess Gibbs free to form more stable mixed monolayer. Among the ten components in the sea salts, Ca ions played the major role in condensation of mixed monolayer. The condensed characteristic of the mixed SA (or PA)/DEHP monolayers suggested that the hydrocarbon chains were ordered on artificial seawater. By means of infrared reflection-absorption spectroscopy (IRRAS), we found that multiple sea salt mixtures induced deprotonated forms of fatty acids at the air-water interface. Sea salts can improve the stability and lifetime of mixed fatty and phthalate ester monolayer on aqueous droplets in the atmosphere. Interfacial properties of mixed fatty and phthalate ester monolayers at the air-ocean interface are important to help understand their behavior and fate in the marine environment.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Enhanced Fatty Scavenging and Glycerophospholipid Metabolism Accompany Melanocyte Neoplasia Progression in Zebrafish.

Alterations in lipid metabolism in cancer cells impact cell structure, signaling, and metabolism, making lipid metabolism a potential diagnostic marker and therapeutic target. In this study, we combined PET, desorption electrospray ionization-mass spectrometry (DESI-MS), nonimaging MS, and transcriptomic analyses to interrogate changes in lipid metabolism in a transgenic zebrafish model of oncogenic RAS-driven melanocyte neoplasia progression. Exogenous fatty uptake was detected in melanoma tumor nodules by PET using the surrogate tracer 14(R,S)-18F-fluoro-6-thia-heptadecanoic ([18F]-FTHA), consistent with upregulation of genes associated with fatty uptake found through microarray analysis. DESI-MS imaging revealed that FTHA uptake in tumors was heterogeneous. Transcriptome and lipidome analyses further highlighted dysregulation of glycerophospholipid pathways in melanoma tumor nodules, including increased abundance of phosphatidyl ethanolamine and phosphatidyl choline species, corroborated by DESI-MS, which again revealed heterogeneous phospholipid composition in tumors. Overexpression of the gene encoding lipoprotein lipase (LPL), which was upregulated in zebrafish melanocyte tumor nodules and expressed in the majority of human melanomas, accelerated progression of oncogenic RAS-driven melanocyte neoplasia in zebrafish. Depletion or antagonism of LPL suppressed human melanoma cell growth; this required simultaneous fatty synthase (FASN) inhibition when FASN expression was also elevated. Collectively, our findings implicate fatty acquisition as a possible therapeutic target in melanoma, and the methods we developed for monitoring fatty uptake have potential for diagnosis, patient stratification, and monitoring pharmacologic response. SIGNIFICANCE: These findings demonstrate the translational potential of monitoring fatty uptake and identify lipoprotein lipase as a potential therapeutic target in melanoma.©2019 American Association for Cancer Research.

Keyword: energy

Utilization of lactic in human myotubes and interplay with glucose and fatty metabolism.

Once assumed only to be a waste product of anaerobe glycolytic activity, lactate is now recognized as an source in skeletal muscles. While lactate metabolism has been extensively studied in vivo, underlying cellular processes are poorly described. This study aimed to examine lactate metabolism in cultured human myotubes and to investigate effects of lactate exposure on metabolism of oleic and glucose. Lactic , fatty and glucose metabolism were studied in myotubes using [C(U)]lactic , [C]oleic and [C(U)]glucose, respectively. Myotubes expressed both the MCT1, MCT2, MCT3 and MCT4 lactate transporters, and lactic was found to be a substrate for both glycogen synthesis and lipid storage. Pyruvate and inhibited lactic oxidation, whilst glucose and α-cyano-4-hydroxycinnamic inhibited lactic uptake. Acute addition of lactic inhibited glucose and oleic oxidation, whereas oleic uptake was increased. Pretreatment with lactic for 24\u2009h did not affect glucose or oleic metabolism. By replacing glucose with lactic during the whole culturing period, glucose uptake and oxidation were increased by 2.8-fold and 3-fold, respectively, and oleic oxidation was increased 1.4-fold. Thus, lactic has an important role in metabolism of human myotubes.

Keyword: energy

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on metabolism, but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: energy

Homology modeling and docking studies of a Δ9-fatty desaturase from a Cold-tolerant sp. AMS8.

Membrane-bound fatty desaturases perform oxygenated desaturation reactions to insert double bonds within fatty acyl chains in regioselective and stereoselective manners. The Δ9-fatty desaturase strictly creates the first double bond between C9 and 10 positions of most saturated substrates. As the three-dimensional structures of the bacterial membrane fatty desaturases are not available, relevant information about the enzymes are derived from their amino sequences, site-directed mutagenesis and domain swapping in similar membrane-bound desaturases. The cold-tolerant sp. AMS8 was found to produce high amount of monounsaturated fatty acids at low temperature. Subsequently, an active Δ9-fatty desaturase was isolated and functionally expressed in . In this paper we report homology modeling and docking studies of a Δ9-fatty desaturase from a Cold-tolerant sp. AMS8 for the first time to the best of our knowledge. Three dimensional structure of the enzyme was built using MODELLER version 9.18 using a suitable template. The protein model contained the three conserved-histidine residues typical for all membrane-bound desaturase catalytic activity. The structure was subjected to minimization and checked for correctness using Ramachandran plots and ERRAT, which showed a good quality model of 91.6 and 65.0%, respectively. The protein model was used to preform MD simulation and docking of using CHARMM36 force field in GROMACS Version 5 and Autodock tool Version 4.2, respectively. The docking simulation with the lowest binding , -6.8 kcal/mol had a number of residues in close contact with the docked namely, Ile26, Tyr95, Val179, Gly180, Pro64, Glu203, His34, His206, His71, Arg182, Thr85, Lys98 and His177. Interestingly, among the binding residues are His34, His71 and His206 from the first, second, and third conserved histidine motif, respectively, which constitute the active site of the enzyme. The results obtained are in compliance with the activity of the Δ9-fatty desaturase on the membrane phospholipids.

Keyword: energy

AMPK regulates metabolism through the SIRT1 signaling pathway to improve myocardial hypertrophy.

We investigated the correlations of adenosine monophosphate-activated protein kinase (AMPK), Silence information regulator 1 (SIRT1) and metabolism with myocardial hypertrophy.Myocardial hypertrophy experimental model was established via transverse aortic constriction (TAC)-induced myocardial hypertrophy and phenylephrine (PE)-induced hypertrophic myocardial cell culture. After activation of AMPK, the messenger ribonucleic (mRNA) expressions in myocardial tissue- and myocardial cell hypertrophy-related genes, atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MHC), were detected. The production rate of 14C-labeled 14CO2 from was quantitatively determined to detect the fatty and glucose oxidation of hypertrophic myocardial tissues or cells, and the glucose uptake of myocardial cells was studied using [14C] glucose. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to detect the changes in SIRT1 mRNA and protein expressions in hypertrophic myocardial tissues. Moreover, SIRT1 small interfering ribonucleic (siRNA) was used to interfere in SIRT1 expression to further investigate the role of SIRT1 in the effect of AMPK activation on myocardial hypertrophy.AMPK activation could significantly reduce the mRNA expressions of ANP and β-MHC in vitro and in vivo. AMPK could increase the ejection fraction (EF) and decrease the protein synthesis rate in myocardial cells in mice with myocardial hypertrophy. Besides, AMPK activation could increase the fatty oxidation, improve the glucose uptake and reduce the glucose oxidation. After AMPK activation, both SIRT1 mRNA and protein expressions in hypertrophic myocardial tissues and myocardial cells were increased. After SIRT1 siRNA was further used to interfere in SIRT1 expression in myocardial cells, it was found that mRNA expressions and protein synthesis rates of ANP and β-MHC were increased.The activation of AMPK can inhibit the myocardial hypertrophy, which may be realized through regulating the myocardial metabolism via SIRT1 signaling pathway.

Keyword: energy

and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty constituents are , oleic, and linoleic ), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk fat yield and milk output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable intake, increased milk fat yield and milk output, increased plasma triacylglycerides and long-chain fatty concentrations, and had no effect on retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable intake, and total milk, fat, protein, and lactose yields. Milk output increased at the HMP level, and protein level did not affect total retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra intake into milk and had no effect on milk N efficiency.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

(L.) L. subsp. .: Nutritional Characterization.

The habit of eating wild plants in Europe is often associated with times of famine; an example of such is the nectar of (L.) L., a parasitic plant. To the authors\' best knowledge, there are no studies on its nutritional and chemical composition; thus, the whole (L.) L. subsp. Wettst. plant (CH) and its nectar (NCH) were nutritionally and chemically characterized. The proximate composition of CH and NCH were very similar in terms of , ash, and carbohydrate content. Protein and fat were approximately 2-fold higher in NCH, and crude fiber was 4.6-fold higher in CH compared to NCH. Fructose, glucose, sucrose, and trehalose were the free sugars present in both samples. Oxalic, malic, and citric acids were the identified organic acids in both samples, with citric as the most abundant molecule. For both samples, polyunsaturated and saturated fatty acids (PUFA and SFA, respectively) predominate over monounsaturated fatty acids (MUFA) due to the significant contribution of linoleic and acids, respectively. However, unsaturated fatty acids (UFA) prevail over SFA in CH and NCH. Therefore, CH proved to be an excellent source of nutritional compounds, which supports its use during past periods of scarcity.

Keyword: energy

High-fat diet feeding and increase CRC growth in β2AR-dependent manner.

Epidemiology studies indicate that consumption of high-fat diet (HFD) is directly associated with the development of colorectal cancer (CRC). However, the exact component in HFD and the mechanism underlying its effect on CRC growth remained unclear. Our study shows that HFD feeding increases β2AR expression in the xenograft tissues of CRC-bearing mouse model; the elevated β2AR expression is reduced when HFD is replaced by control diet, which strongly suggests an association between HFD feeding and β2AR expression in CRC. HFD feeding increases and stearic levels in CRC; however, only increases β2AR expression, which is dependent upon Sp1. β2AR plays the dominant role in promoting CRC cell proliferation among all the β-AR subtypes. More importantly, knockout of β2AR or knockdown of Sp1 abolishes the increased CRC cell proliferation, suggesting increases CRC cell proliferation in β2AR-dependent manner. HFD or -rich diet (PAD) also fails to increase the tumor growth in xenograft mouse models bearing β2AR-knockout CRC cells. β2AR promotes CRC growth by increasing the phosphorylation of HSL at the residue S552. The phosphorylated and activated HSL (S552) changes the metabolic phenotype of CRC and increases production, which promotes CRC growth. Our study has revealed the unique tumorigenic properties of in promoting CRC growth, and have delineated the underlying mechanism of action. We are also the first to report the linkage between HFD feeding and β-adrenergic signaling pathway in relation to CRC growth.

Keyword: energy

Fatty composition in serum cholesterol esters and phospholipids is linked to visceral and subcutaneous adipose tissue content in elderly individuals: a cross-sectional study.

Visceral adipose tissue (VAT) and truncal fat predict cardiometabolic disease. Intervention trials suggest that saturated fatty acids (SFA), e.g. , promote abdominal and liver fat storage whereas polyunsaturated fatty acids (PUFA), e.g. linoleic , prevent fat accumulation. Such findings require investigation in population-based studies of older individuals. We aimed to investigate the relationships of serum biomarkers of PUFA intake as well as serum levels of , with abdominal and total adipose tissue content.In a population-based sample of 287 elderly subjects in the PIVUS cohort, we assessed fatty composition in serum cholesterol esters (CE) and phospholipids (PL) by gas chromatography and the amount of VAT and abdominal subcutaneous (SAT) adipose tissue by magnetic resonance imaging (MRI), liver fat by MR spectroscopy (MRS), and total body fat, trunk fat and leg fat by dual- X-ray absorptiometry (DXA). Insulin resistance was estimated by HOMA-IR.VAT and trunk fat showed the strongest correlation with insulin resistance (r\u2009=\u20090.49, P\u2009<\u20090.001). Linoleic in both CE and PL was inversely related to all body fat depots (r\u2009=\u2009-0.24 to -0.33, P\u2009<\u20090.001) including liver fat measured in a sub-group (r\u2009=\u2009-0.26, P\u2009<\u20090.05, n\u2009=\u200973), whereas n-3 PUFA showed weak inverse (18:3n-3) or positive (20:5n-3) associations. in CE, but not in PL, was directly correlated with VAT (r\u2009=\u20090.19, P\u2009<\u20090.001) and trunk fat (r\u2009=\u20090.18, P\u2009=\u20090.003). Overall, the significant associations remained after adjusting for intake, height, alcohol, sex, smoking, education and physical activity. The inverse correlation between linoleic and VAT remained significant after further adjustment for total body fat.Serum linoleic is inversely related to body fat storage including VAT and trunk fat whereas was less consistently but directly associated, in line with recent feeding studies. Considering the close link between VAT and insulin resistance, a potential preventive role of plant-based PUFA in VAT accumulation warrants further study.

Keyword: energy

Apoptotic germ cells regulate Sertoli cell lipid storage and fatty oxidation

The presence of lipid droplets (LDs) and the utilization of fatty acids (FAs) as a source of are Sertoli cell (SC) putative characteristics. It is well known that SCs can phagocyte and degrade apoptotic germ cells (AGCs) resulting in increasing lipid content and ATP levels. A relationship between the regulation of lipid storage and of lipid oxidation in SC might be envisaged. The aim of this study was to analyze whether AGCs and FAs are able to simultaneously regulate molecular mechanisms involved in lipid storage and in FA oxidation in SC. The experimental model utilized in this study consisted in SC cultures obtained from 20-day-old rats that were co-cultured with AGC or treated with (PA, 500 μM) for 24 and 48 h. AGC and PA increase LD, triacylglycerol (TAG) content and mRNA levels of Plin1, Plin2, Plin3 (proteins involved in TAG storage). Simultaneously, AGC and PA rise the extent of FA oxidation and mRNA levels of Cpt1 and Lcad (proteins involved in FA degradation). Results also show that peroxisome proliferator-activated receptor (PPAR) transcriptional activity, transcription factor which participate in lipid metabolism regulation, increases by AGC and PA treatment in SC. Additionally, the presence of a PPARg antagonist decreases the upregulation of LD content and Plin1 expression. Similarly, the presence of a PPARb/d antagonist reduces the increase in FA oxidation and Cpt1 mRNA levels. Altogether these results suggest that AGC and FA, which probably generate PPAR ligands, regulate lipid storage and fatty utilization, contributing to the homeostasis in the seminiferous tubules.

Keyword: energy

Calculation of apparent pK values of saturated fatty acids with different lengths in DOPC phospholipid bilayers.

We performed all-atom molecular dynamics simulations and calculated free profiles and apparent pKa values for neutral and anionic forms of single myristic (C14:0), (C16:0) and stearic (C18:0) fatty embedded in a DOPC bilayer and explicit water solvent. We showed that the neutral forms of the fatty acids are stabilized inside the bilayer by hydrogen bonding of a fatty carboxylic group with DOPC phosphate and carbonyl groups. In contrast to the neutral form, the anionic forms of the fatty acids are shifted towards the water-membrane interface and are instead stabilized by hydrogen bonding to interfacial water. By using umbrella sampling simulations, we calculated free energies of stabilization and revealed that the free of stabilization inside the bilayer increases with the chain length for both the neutral and deprotonated forms. On the other hand, the free energies of flip-flop of both the neutral and anionic forms are constant upon the prolongation of the fatty . Based on the free curves, we also calculated apparent fatty pKa,app values in the bilayer, which are 7.0, 7.2 and 6.3 for myristic, and stearic and are increased by several pKa units compared to the corresponding pKa values in water. By further analysis of the calculated curves we found that spontaneous protonation of fatty anions takes place in the bilayer interior at ca. 1.4 nm from the bilayer center for all studied fatty acids.

Keyword: energy

Selective synthesis of human milk fat-style structured triglycerides from microalgal oil in a microfluidic reactor packed with immobilized lipase.

Human milk fat-style structured triacylglycerols were produced from microalgal oil in a continuous microfluidic reactor packed with immobilized lipase for the first time. A remarkably high conversion efficiency was demonstrated in the microreactor with reaction time being reduced by 8 times, Michaelis constant decreased 10 times, the lipase reuse times increased 2.25-fold compared to those in a batch reactor. In addition, the content of at sn-2 position (89.0%) and polyunsaturated fatty acids at sn-1, 3 positions (81.3%) are slightly improved compared to the product in a batch reactor. The increase of melting points (1.7°C) and decrease of crystallizing point (3°C) implied higher quality product was produced using the microfluidic technology. The main cost can be reduced from $212.3 to $14.6 per batch with the microreactor. Overall, the microfluidic bioconversion technology is promising for modified functional lipids production allowing for cost-effective approach to produce high-value microalgal coproducts.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: energy

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: energy

Loss of biliverdin reductase-A promotes lipid accumulation and lipotoxicity in mouse proximal tubule cells.

Obesity and increased lipid availability have been implicated in the development and progression of chronic kidney disease. One of the major sites of renal lipid accumulation is in the proximal tubule cells of the kidney, suggesting that these cells may be susceptible to lipotoxicity. We previously demonstrated that loss of hepatic biliverdin reductase A (BVRA) causes fat accumulation in livers of mice on a high-fat diet. To determine the role of BVRA in mouse proximal tubule cells, we generated a CRISPR targeting BVRA for a knockout in mouse proximal tubule cells (BVRA KO). The BVRA KO cells had significantly less metabolic potential and mitochondrial respiration, which was exacerbated by treatment with , a saturated fatty . The BVRA KO cells also showed increased intracellular triglycerides which were associated with higher fatty uptake gene cluster of differentiation 36 as well as increased de novo lipogenesis as measured by higher neutral lipids. Additionally, neutrophil gelatinase-associated lipocalin 1 expression, annexin-V FITC staining, and lactate dehydrogenase assays all demonstrated that BVRA KO cells are more sensitive to -induced lipotoxicity than wild-type cells. Phosphorylation of BAD which plays a role in cell survival pathways, was significantly reduced in -treated BVRA KO cells. These data demonstrate the protective role of BVRA in proximal tubule cells against saturated fatty -induced lipotoxicity and suggest that activating BVRA could provide a benefit in protecting from obesity-induced kidney injury.

Keyword: energy

Association of dietary factors with severity of coronary artery disease.

Cardiovascular disease has taken epidemic proportions during past decades. Cardiovascular risk factors contribute to progression of coronary lesions, worsening the patient\'s prognosis. This study was planned to analyze the association of dietary factors with severity of coronary artery disease (CAD) in Indian patients.Three hundred patients with known coronary disease above the age of 25 years were included in this study. Blood samples were collected for biochemical markers. Patients were stratified according to severity of CAD [number of vessel involved-single (SVD), double (DVD), triple (TVD)].Mean age of the patient was 60.9\xa0±\xa012.4 years. Subjects with TVD, DVD, SVD in the study were 52.3%, 25.3% and 22.3% respectively. Patients with TVD had higher body mass index, triglycerides, HOMA-Insulin Resistance, hsCRP and lower high density cholesterol. Diabetes mellitus, hypertension and dyslipidemia were more common in TVD patients. Among macronutrients, patients with TVD had higher intake of carbohydrate and lower intake of protein and dietary fibers. There was no association of total fat intake with CAD, however, intake of was higher among patients with TVD. Intake of vitamins namely niacin, riboflavin, thiamine, B6, and vitamin-C decreased with increase in severity. With increase in severity of CAD, mineral intake (potassium, calcium, magnesium, phosphorus, sulfur, iron, chromium, copper, manganese, and zinc) decreased.Dietary factors are associated with severity of coronary artery disease. Low intake of protein, fiber, vitamins, minerals and high intake of carbohydrate and fat was associated with higher probability of having severe CAD.Copyright © 2016 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.

Keyword: energy

Valorization of sewage sludge via non-catalytic transesterification.

To seek a way to valorize sewage sludge (SS), it was chosen as a raw material for biodiesel production. As such, non-catalytic transesterification of dried SS was carried out, to enhance its value. Note that picking a waste material such as SS as an inexpensive lipid feedstock for biodiesel production, without lipid extraction, greatly increases the economic viability of biodiesel. Also, to enhance biodiesel sustainability, ethanol (EtOH) was employed as the acyl acceptor, in this study, and this was experimentally justified by results showing that employing EtOH as an acyl acceptor provided an effective means for compensating for the lower heating value arising from the large amount of (C) in SS. This study experimentally proved that the fatty ethyl ester (FAEE) yield at 380\u202f°C reached up to 13.33\u202fwt% of dried SS. Given that the lipid content of dried SS is 14.01\u202f±\u202f0.64\u202fwt%, the FAEE yield of 13.33\u202fwt% implied that 95.14\u202fwt% of lipid in dried SS had been converted into FAEEs. The introduced SS valorization in this study offered an excellent opportunity to address diverse environmental hazards arising from SS and associated emerging contaminants. Given that the optimal temperature for the non-catalytic conversion for biodiesel production from SS was found to be 380\u202f°C, emerging contaminants, such as microplastics and antimicrobials, were simultaneously degraded, due to their inferior thermal stabilities. Lastly, considering that the introduced biodiesel conversion process is thermally induced, the SS reside after the biodiesel conversion process can be further used in thermo-chemical processes as raw materials for gasification and pyrolysis (future work).Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.

Keyword: energy

Gene expressions and metabolomic research on the effects of polyphenols from the involucres of Castanea mollissima Blume on heat-stressed broilers chicks.

To study the effects of polyphenolic extract from involucres of Castanea mollissima Blume ( PICB: ), a novel approach using gene expression by real time polymerase chain reaction ( REAL-TIME PCR: ) coupled with metabolomic profiling technique was established to explain the mechanism of PICB on heat-stressed broiler chicks. Four thousand 28-day-old male Arbor Acres (AA) broilers were randomly assigned to 5 groups (4 replicates / group, 20 chicks / replicate), in which group 1 was normal control group fed with basic ration; groups 2, 3, 4, and 5 were fed with the basic ration with a supplementation of 0.2% Vitamin C ( VC: ), or 0.2%, 0.3%, or 0.4% of PICB respectively. After 1 wk of adaptation, heat stress was applied for 7 consecutive days. On d 3 and d 7 of heat stress, the chicks were sacrificed and sampled. The mRNA expression of heat stress protein 70 (HSP70), glutathione peroxidase ( GSH-PX: ), ornithine decarboxylase ( ODC: ), epidermal growth factor ( EGF: ) and epidermal growth factor receptor ( EGFR: ) were detected by real-time PCR using samples from jejunum mucosa. The serum and jejunum mucosa metabolomic profiles of PICB group showing best antioxidative effects and control group at d 3 were studied using the method of the gas chromatography - time of flight mass spectrometry ( GT-TOF-MS: ), followed by principal component analysis and partial least squares-discriminate analysis. Potential biomarkers were found using Student\'s t-test. The results showed mRNA expressions of HSP70, GSH-Px, ODC, EGF, and EGFR were altered by the supplementation of PICB. PICB exhibited antioxidative and growth promoting effects, and 0.3% PICB supplementation level exhibited the best. Three metabolites in the serum and 5 in the jejunum mucosa were identified as potential biomarkers. They were considered to be in accordance with antioxidative and growth promoting effects of PICB, which involved in the metabolism (sorbitol, ), carbohydrate metabolism, amino acids metabolism (serine, L-ornithine), glutathione metabolism (glutamate, L-ornithine), GnRH signaling pathway (inositol), etc. These findings provided novel insights into our understanding of molecular mechanism of PICB effects on heat-stressed chicks.© 2016 Poultry Science Association Inc.

Keyword: energy

DGAT1-Dependent Lipid Droplet Biogenesis Protects Mitochondrial Function during Starvation-Induced Autophagy.

Lipid droplets (LDs) provide an "on-demand" source of fatty acids (FAs) that can be mobilized in response to fluctuations in nutrient abundance. Surprisingly, the amount of LDs increases during prolonged periods of nutrient deprivation. Why cells store FAs in LDs during an crisis is unknown. Our data demonstrate that mTORC1-regulated autophagy is necessary and sufficient for starvation-induced LD biogenesis. The ER-resident diacylglycerol acyltransferase 1 (DGAT1) selectively channels autophagy-liberated FAs into new, clustered LDs that are in close proximity to mitochondria and are lipolytically degraded. However, LDs are not required for FA delivery to mitochondria but instead function to prevent acylcarnitine accumulation and lipotoxic dysregulation of mitochondria. Our data support a model in which LDs provide a lipid buffering system that sequesters FAs released during the autophagic degradation of membranous organelles, reducing lipotoxicity. These findings reveal an unrecognized aspect of the cellular adaptive response to starvation, mediated by LDs.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Effects of timing of supplementation during early lactation on nutrient digestibility, balance, and metabolism of dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation during early lactation on nutrient digestibility, intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible intake by 3.9 Mcal/d, metabolizable intake by 3.5 Mcal/d, and net for lactation intake by 2.5 Mcal/d. The PA diet also increased milk output, negative balance, and plasma nonesterified fatty concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for balance due to cows receiving the PA treatment being in a greater negative balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased intake and milk output and did not affect balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and intake compared with a control diet containing no supplemental fat. Feeding C16:0 markedly increased milk output in both the FR and PK periods but increased negative balance only in the FR period.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Metabolic transition of milk triacylglycerol synthesis in response to varying levels of palmitate in porcine mammary epithelial cells.

Milk in mammals is a key source of lipids for offspring, providing both critical and essential fatty acids. For lactating sows, is one of the most abundant fatty acids in milk, providing 10~12% of the suckling pig total dietary supply. However, the effects of exogenous on milk fat synthesis in sow mammary glands are not well-known. In this study, we investigated the effects of on lipogenic genes in porcine mammary epithelial cells (pMECs) to explore the role of exogenous in mediating milk triacylglycerols (TAG) synthesis.Porcine mammary epithelial cells were cultured for 24\xa0h in the presence of different concentrations of palmitate (0, 25, 50, 100, 200, 400, and 600\xa0μM). The effect of palmitate on cell viability was tested via MTT assay. Intracellular lipid accumulation was measured through Oil Red O staining, and TAG levels were quantified by enzymatic colorimetric methods. Expression of genes and proteins involved in milk fat biosynthesis were assayed with quantitative real-time polymerase chain reaction (PCR) and Western blotting, respectively.Incubation with palmitate promoted cellular lipid synthesis in a dose-dependent manner, as reflected by the increased TAG content and enhanced formation of cytosolic lipid droplets. The increased lipid synthesis by palmitate was probably attributable to the upregulated mRNA expression of genes associated with milk fat biosynthesis, including long-chain fatty uptake (, ), intracellular activation and transport , ), TAG synthesis (, , ), lipid droplet formation (), and regulation of transcription (). Western blot analysis of CD36 and DGAT1 proteins confirmed the increased lipid synthesis with increasing incubation of palmitate. However, the genes involved in fatty de novo synthesis (, ), fatty desaturation (), and regulation of transcription (, ) were inversely affected by incubation with increasing concentrations of palmitate. Western blot analysis of ACACA protein confirmed this decrease associated with increasing levels of palmitate.Results from this study suggest that palmitate stimulated the cytosolic TAG accumulation in pMECs, probably by promoting lipogenic genes and proteins that are involved in lipid synthesis. However, addition of palmitate decreased the fatty de novo synthesis in pMECs.

Keyword: energy

Effects of timing of supplementation on production responses of early-lactation dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a diet supplemented with C16:0 (, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% fat-corrected milk by 5.30 kg/d, yield of -corrected milk (ECM) by 4.70 kg/d, milk fat content by 0.41% units, milk fat yield by 280 g/d, and protein yield by 100 g/d. The increase in milk fat associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased body weight (BW) by 21 kg and body condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk fat yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% fat-corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk fat content by 0.22% units, milk fat yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk fat yield due to feeding PA during the PK period increasing milk fat yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive oxygen species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect inflammation and lipid content in the liver.The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and inflammation induced by HF diet , leading to improved metabolic disorder and associated liver injury in a mouse model of non-alcoholic fatty liver disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: energy

Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

A total of ten marine yeast strains isolated from Bohai Sea, Northern China were identified to be members of three genera Rhodosporidium, Rhodotorula, and Cryptococcus. Two representative strains Rhodosporidium TJUWZ4 and Cryptococcus TJUWZA11 with high lipid content based on Nile red staining method were further characterized. A wide range of culture conditions (C and N sources, pH, temperature, salinity and C/N ratio) were tested to characterize the biomass and lipid production (yield and productivity) of these strains. Results indicated that Rhodosporidium TJUWZ4 was capable of achieving lipid yield up to 44% and 0.09\xa0g/l-h productivity on glucose and peptone medium at pH 4, 20\xa0°C, 30% salinity, and C/N 80. Three fatty acids, namely oleic (18:1), (C16:0) and linoleic (18:2) were the major intracellular fatty acids, which accounted for 90% of total lipids. With promising features for intracellular lipid accumulation, Rhodosporidium TJUWZ4 is a robust strain with great potentials for application in biodiesel production from renewable feedstocks.

Keyword: energy

Ecological relevance of skeletal fatty concentration and composition in Mediterranean scleractinian corals.

The intra-skeletal fatty concentration and composition of four Mediterranean coral species, namely Cladocora caespitosa, Balanophyllia europaea, Astroides calycularis and Leptopsammia pruvoti, were examined in young and old individuals living in three different locations of the Mediterranean Sea. These species are characterized by diverse levels of organization (solitary or colonial) and trophic strategies (symbiotic or non-symbiotic). Fatty acids have manifold fundamental roles comprehensive of membrane structure fluidity, cell signaling and storage. For all species, except for B. europaea, the intra-skeletal fatty concentration was significantly higher in young individuals than in old ones. Moreover, fatty concentration was higher in colonial corals than in solitary ones and in the symbiotic corals compared to non-symbiotic ones. Analysis by gas chromatography-mass spectrometry (GC-MS) revealed that (16:0) was the most abundant fatty , followed by stearic (18:0) in order of concentration. Oleic (18:1) was detected as the third main component only in skeletons from symbiotic corals. These results suggest that, in the limits of the studied species, intra-skeletal fatty composition and concentration may be used for specific cases as a proxy of level of organization and trophic strategy, and eventually coral age.

Keyword: energy

Hydroxyhydroquinone impairs fat utilization in mice by reducing nitric oxide availability.

Habitual consumption of chlorogenic compounds (CGAs) from coffee increases fat catabolism and reduces body fat; however, the contribution of roasted coffee remains unclear. Hydroxyhydroquinone (HHQ) impairs the vasodilatory and antihypertensive effects of CGAs by reducing nitric oxide (NO) bioavailability. Since HHQ also reduces fat catabolism, we hypothesized that HHQ does so by decreasing NO availability. Therefore, we investigated the effect of HHQ on metabolism in KKAy mice. In HHQ-treated mice, fat oxidation was significantly low and dose-dependent, serum and urinary hydrogen peroxide were high, and plasma NO metabolites and S-nitrosylated liver proteins were low. In HHQ-treated mouse hepatocytes, the palmitate-induced increase in cellular oxygen consumption was negatively affected, and HHQ or L-NAME reduced cellular fatty utilization. In conclusion, HHQ can impair fat utilization by reducing NO availability in mice. Protein S-nitrosylation reduction in liver cells after HHQ consumption may be associated with impaired fatty oxidation.

Keyword: energy

Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.

Mitochondrial trafficking plays a central role in dorsal root ganglion (DRG) neuronal cell survival and neurotransmission by transporting mitochondria from the neuronal cell body throughout the bundles of DRG axons. In type 2 diabetes (T2DM), dyslipidemia and hyperglycemia damage DRG neurons and induce mitochondrial dysfunction; however, the impact of free fatty acids and glucose on mitochondrial trafficking in DRG neurons remains unknown. To evaluate the impact of free fatty acids compared to hyperglycemia on mitochondrial transport, primary adult mouse DRG neuron cultures were treated with physiologic concentrations of palmitate and glucose and assessed for alterations in mitochondrial trafficking, mitochondrial membrane potential, and mitochondrial bioenergetics. Palmitate treatment significantly reduced the number of motile mitochondria in DRG axons, but physiologic concentrations of glucose did not impair mitochondrial trafficking dynamics. Palmitate-treated DRG neurons also exhibited a reduction in mitochondrial velocity, and impaired mitochondrial trafficking correlated with mitochondrial depolarization in palmitate-treated DRG neurons. Finally, we found differential bioenergetic effects of palmitate and glucose on resting and energetically challenged mitochondria in DRG neurons. Together, these results suggest that palmitate induces DRG neuron mitochondrial depolarization, inhibiting axonal mitochondrial trafficking and altering mitochondrial bioenergetic capacity.-Rumora, A. E., Lentz, S. I., Hinder, L. M., Jackson, S. W., Valesano, A., Levinson, G. E., Feldman, E. L. Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.© FASEB.

Keyword: energy

Disruption of Lipid Uptake in Astroglia Exacerbates Diet-Induced Obesity.

Neuronal circuits in the brain help to control feeding behavior and systemic metabolism in response to afferent nutrient and hormonal signals. Although astrocytes have historically been assumed to have little relevance for such neuroendocrine control, we investigated whether lipid uptake via lipoprotein lipase (LPL) in astrocytes is required to centrally regulate homeostasis. Ex vivo studies with hypothalamus-derived astrocytes showed that LPL expression is upregulated by oleic , whereas it is decreased in response to or triglycerides. Likewise, astrocytic LPL deletion reduced the accumulation of lipid droplets in those glial cells. Consecutive in vivo studies showed that postnatal ablation of LPL in glial fibrillary acidic protein-expressing astrocytes induced exaggerated body weight gain and glucose intolerance in mice exposed to a high-fat diet. Intriguingly, astrocytic LPL deficiency also triggered increased ceramide content in the hypothalamus, which may contribute to hypothalamic insulin resistance. We conclude that hypothalamic LPL functions in astrocytes to ensure appropriately balanced nutrient sensing, ceramide distribution, body weight regulation, and glucose metabolism.© 2017 by the American Diabetes Association.

Keyword: energy

Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon source.

Industrial biotechnology that is able to provide environmentally friendly bio-based products has attracted more attention in replacing petroleum-based industries. Currently, most of the carbon sources used for fermentation-based bioprocesses are obtained from agricultural commodities that are used as foodstuff for human beings. Lignocellulose-derived sugars as the non-food, green, and sustainable alternative carbon sources have great potential to avoid this dilemma for producing the renewable, bio-based hydrocarbon fuel precursors, such as microbial lipid. Efficient bioconversion of lignocellulose-based sugars into lipids is one of the critical parameters for industrial application. Therefore, the\xa0fed-batch cultivation, which is a common method used in industrial applications, was investigated to achieve a high cell\xa0density culture along with high lipid yield and productivity.In this study, several fed-batch strategies were explored to improve lipid production using lignocellulosic hydrolysates derived from corn stover. Compared to the batch culture giving a lipid yield of 0.19\xa0g/g, the dissolved-oxygen-stat feeding mode increased the lipid yield to 0.23\xa0g/g and the lipid productivity to 0.33\xa0g/L/h. The pulse feeding mode further improved lipid productivity to 0.35\xa0g/L/h and the yield to 0.24\xa0g/g. However, the highest lipid yield (0.29\xa0g/g) and productivity (0.4\xa0g/L/h) were achieved using an automated online sugar control feeding mode, which gave a dry cell weight of 54\xa0g/L and lipid content of 59\xa0% (w/w). The major fatty acids of the lipid derived from lignocellulosic hydrolysates were predominately and oleic , which are similar to those of conventional oilseed plants.Our results suggest that the fed-batch feeding strategy can strongly influence the lipid production. The online sugar control feeding mode was the most appealing strategy for high cell density, lipid yield, and lipid productivity using lignocellulosic hydrolysates as the sole carbon source.

Keyword: energy

Changes of albumin secondary structure after binding. FT-IR spectroscopic study.

Albumin is an universal transport protein. Plasma pool of free fatty acids arising from triglyceride hydrolysis, critical in metabolism and etiology of metabolic disorders is transported by albumin. According to various studies albumin has from seven to nine binding sites with diverse affinity to long chain fatty acids. X-ray diffraction crystallography measurements have provided data only for pure human serum albumin or albumin with fully saturated binding sites. These results have shown that amount of \uf061-helices is higher after fatty acids binding. Molecular mechanics simulations suggest that binding of fatty acids in two high-affinity sites leads to major conformational changes in albumin structure. The aim of this research was to investigate albumin secondary structure upon gradually increasing fatty acids to protein mole ratio.Fourier transform infrared spectroscopy was applied to study changes of bovine serum albumin (as an analogue of human serum albumin) \uf061-helical structures after binding in a range of 0-20 : albumin molar ratios representing pure protein, partial, full saturation and excess binding sites capacity.Amount of \uf061-helices was increasing along with the amount of : bovine serum albumin molar ratio and reached maximum value around 2 mol/mol.Our studies confirmed molecular mechanics simulations and crystallographic studies. binding in two high-affinity sites leads to major structural changes, filling another sites only slightly influenced bovine serum albumin secondary structure. The systematic study of fatty acids and albumin interactions, using an experimental model mimicking metabolic disorders, may results in new tools for personalized nanopharmacotherapy.

Keyword: energy

Exposure of bovine oocytes and embryos to elevated non-esterified fatty concentrations: integration of epigenetic and transcriptomic signatures in resultant blastocysts.

Metabolic stress associated with negative balance in high producing dairy cattle and obesity in women is a risk factor for decreased fertility. Non-esterified fatty acids (NEFA) are involved in this pathogenesis as they jeopardize oocyte and embryo development. Growing evidence indicates that maternal metabolic disorders can disturb epigenetic programming, such as DNA methylation, in the offspring. Oocyte maturation and early embryo development coincide with methylation changes and both are sensitive to adverse environments. Therefore, we investigated whether elevated NEFA concentrations affect establishment and maintenance of DNA methylation in oocytes and embryos, subsequently altering transcriptomic profiles and developmental competence of resultant blastocysts.Bovine oocytes and embryos were exposed to different NEFA concentrations in separate experiments. In the first experiment, oocytes were matured in vitro for 24\xa0h in medium containing: 1) physiological ("BASAL") concentrations of oleic (OA), (PA) and stearic (SA) or 2) pathophysiological ("HIGH COMBI") concentrations of OA, PA and SA. In the second experiment, zygotes were cultivated in vitro for 6.5\xa0days under BASAL or HIGH COMBI conditions. Developmental competence was evaluated by assessing cleavage and blastocyst rate. Overall gene expression and DNA methylation of resultant blastocysts were analyzed using microarray. DNA methylation data were re-evaluated by pyrosequencing. HIGH COMBI-exposed oocytes and embryos displayed a lower competence to develop into blastocysts compared to BASAL-exposed counterparts (19.3% compared to 23.2% and 18.2% compared to 25.3%, respectively) (P\u2009<\u20090.05). HIGH COMBI-exposed oocytes and embryos resulted in blastocysts with altered DNA methylation and transcriptomic fingerprints, compared to BASAL-exposed counterparts. Differences in gene expression and methylation were more pronounced after exposure during culture compared to maturation suggesting that zygotes are more susceptible to adverse environments. Main gene networks affected were related to lipid and carbohydrate metabolism, cell death, immune response and metabolic disorders.Overall, high variation in methylation between blastocysts made it difficult to draw conclusions concerning methylation of individual genes, although a clear overview of affected pathways was obtained. This may offer clues regarding the high rate of embryonic loss and metabolic diseases during later life observed in offspring from mothers displaying lipolytic disorders.

Keyword: energy

Elevated free fatty acids affect bovine granulosa cell function: a molecular cue for compromised reproduction during negative balance.

High-yielding dairy cows postpartum face the challenge of negative balance leading to elevated free fatty acids levels in the serum and follicular fluid thus affecting the ovarian function. Here, we investigated effects of physiological concentrations of (PA), stearic (SA) and oleic (OA) on the viability, steroid production and gene expression in a bovine granulosa cell (GC) culture model. Treatment with individual and combined fatty acids increased the CD36 gene expression, while no significant apoptotic effects were observed. Both PA and SA significantly upregulated the expression of FSHR, LHCGR, CYP19A1, HSD3B1, CCND2 and increased 17β-estradiol (E2) production, while OA downregulated the expression of these genes and reduced E2. Interestingly, STAR was equally downregulated by all fatty acids and combination treatment. E2 was significantly reduced after combination treatment. To validate the effects of OA, in vivo growing dominant follicles (10-19 mm) were injected with bovine serum albumin (BSA) with/without conjugated OA. The follicular fluid was recovered 48 h post injection. As in our in vitro model, OA significantly reduced intrafollicular E2 concentrations. In addition, expression of CD36 was significantly up- and that of CYP19A1 and STAR significantly downregulated in antral GC recovered from aspirated follicles. The ovulation rates of OA-injected follicles tended to be reduced. Our results indicate that elevated free fatty concentrations specifically target functional key genes in GC both in vitro and in vivo. Suggestively, this could be a possible mechanism through which elevated free fatty acids affect folliculogenesis in dairy cows postpartum.

Keyword: energy

Detection of organic compounds in impact glasses formed by the collision of an extraterrestrial material with the Libyan Desert (Africa) and Tasmania (Australia).

Impact glasses are rich silica melted formed at high temperature and pressure by the impact of an extraterrestrial body on Earth. Here, Libyan Desert glasses (LDGs) and Darwin glasses (DGs) were studied. Two non-destructive analytical techniques were used to detect and characterize organic compounds present in their inclusions: Raman spectroscopy and scanning electron microscopy coupled to -dispersive X-ray spectroscopy (SEM-EDS). Phytoliths, humboldtine, , myristic , oleic , 4-methyl phthalic , and S-H stretching vibrations of amino acids were identified. The presence of these particular organic compounds in such materials has not been reported so far, providing information about (a) the ancient matter of the area where the impact glasses were formed, (b) organic matter belonging to the extraterrestrial body which impacted on the Earth, or (c) even to current plant or bacterial life, which could indicate an active interaction of the LDG and DG with the surrounding environment. Moreover, the identification of fullerene allowed us to know a pressure (15\xa0GPa) and temperatures (670\xa0K or 1800-1900\xa0K) at which samples could be subjected.

Keyword: energy

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating obesity-induced inflammation.

Keyword: energy

α-Ionone attenuates high- diet-induced skeletal muscle wasting in mice via activation of cAMP signaling.

Skeletal muscle atrophy is a common and debilitating condition that lacks a pharmacological therapy. Our aim was to investigate the potential of α-ionone, a naturally occurring flavoring agent, in preventing muscle atrophy and to delineate the mechanisms involved. We found that α-ionone not only stimulated myogenesis but also attenuated -induced atrophy of cultured skeletal myotubes, as evidenced by an increased myotube diameter and length, fusion index, and cellular protein content. These beneficial actions of α-ionone were abrogated by cAMP inhibitor. The antiatrophic effects of α-ionone on cultured myotubes were confirmed in a corresponding mouse model. The skeletal muscle mass, muscle protein content, myofiber diameter, and muscle strength were greater in α-ionone-treated mice than in untreated animals fed high- diet. Furthermore, α-ionone increased cAMP concentration and enhanced its downstream PKA-CREB signaling in skeletal muscle of mice fed high- diet. Thus, α-ionone is a promising agent that may enhance skeletal muscle mass and strength.

Keyword: fat metabolism

Dietary zerumbone from shampoo ginger: new insights into its antioxidant and anticancer activity.

The dietary sesquiterpene dienone zerumbone (ZER) selectively targets cancer cells, inducing mitochondrial dysfunction and apoptosis, and protects non-cancerous cells towards oxidative stress and insult. This study examines the in vitro effects of ZER on peroxidation in biological systems (cholesterol and phospholipid membrane oxidation) and explores its antitumor action in terms of its ability to modulate cancer cell profile. Evaluation of the antioxidant activity of ZER showed that this compound is unable to trap lipoperoxyl radicals per se. ZER significantly modulated the total and fatty profiles in cancer cells, inducing marked changes in the phospholipid/cholesterol ratio, with significant decreases in the levels of oleic and acids and a marked increase of stearic . Cell-based fluorescent measurements of intracellular membranes and droplets using the Nile Red staining technique showed that in cancer cells, ZER induced significant accumulation of cytosolic droplets and altered cell membrane organization/protein dynamics, depolarizing the mitochondrial membranes and inducing apoptosis and alteration of nuclear morphology. The modulatory activity of ZER on the total and fatty profiles and droplets may therefore represent another possible mechanism of its anticancer properties.

Keyword: fat metabolism

Overexpression of Acyl-ACP Thioesterases, and , Induce Distinct Gene Expression Reprogramming in Developing Seeds of .

We examined the substrate preference of acyl-ACP thioesterases, and , and gene expression changes associated with the modification of composition in the seed, using transgenic plants overexpressing or under the control of a seed-specific promoter. seeds contained a higher level of total saturated fatty (FA) content, with 4.3 times increase in 16:0 , whereas seeds showed approximately 3% accumulation of 10:0 and 12:0 medium-chain FAs, and a small increase in other saturated FAs, resulting in higher levels of total saturated FAs. RNA-Seq analysis using entire developing pods at 8, 25, and 45 days after flowering (DAF) showed up-regulation of genes for β-ketoacyl-acyl carrier protein synthase I/II, stearoyl-ACP desaturase, oleate desaturase, and linoleate desaturase, which could increase unsaturated FAs and possibly compensate for the increase in 16:0 at 45 DAF in transgenic plants. In transgenic plants, many putative chloroplast- or mitochondria-encoded genes were identified as differentially expressed. Our results report comprehensive gene expression changes induced by alterations of seed FA composition and reveal potential targets for further genetic modifications.

Keyword: fat metabolism

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on and inflammation in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet metabolic disorder and islet inflammation in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high- diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: fat metabolism

[The becoming of reactions of lipolysis in phylogenesis. The and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V].

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring. In lipoproteins of very low density and lipoproteins of low density, the carnivorous transfer exogenous non-polar triglycerides. The herbivorous also transfer non-polar triglycerides though synthesized by hepatocytes from glucose endogenically. In herbivorous, transferring of non-polar triglycerides prior to synthesis of insulin is forming apoB-100 in composition of lipoproteins of very low density and lipoproteins of low density. The hydrolysis of non-polar triglycerides in lipoproteins of very low density is activated by hepatic glycerol hydrogenase and apoC-III; cells absorb lipoproteins of low density by means of apoB-100 endocytosis. The content on lipoproteins in blood plasma under electrophoresis of lipoproteins corresponds to hepatic glycerol hydrogenase type IIb. In frst and second types of fatty acids transferring in form of triglycerides to lipoproteins of very low density + lipoproteins of low density predominate fatty , triglycerides of the same name and of fatty acids in vivo. The insulin initiated the third type of transferring of oleic fatty by now to insulin-depended cells only in oleic lipoproteins of very low density; hydrolysis of oleic triglycerides is activated by late in phylogenesis post-heparin hepatic glycerol hydrogenase and apoC-II cofactor. The dynamic apoE is actively bound by apoB-100 forming apoE/B-100 ligand. At later stages of phylogenesis insulin formed fatty acids transferring in form of oleic triglycerides in lipoproteins of very low density of the same name without forming of oleic lipoproteins of low density; the electrophoregram of lipoproteins reflects absence of hepatic glycerol hydrogenase. In phylogenesis three types of fatty acids transferring to triglycerides in composition of lipoproteins formed sequentially: 1) chylomicrons + lipoproteins of very low and density + lipoproteins of low density; 2) lipoproteins of very low density + lipoproteins of low density; 3) only in lipoproteins of very low density. The frst one is specifc to piscivorous (carnivorous) while living in ocean. The second one is implemented by herbivorous while they didn\'t begin to synthesize insulin and hepatocytes not yet transform all endogenous fatty into oleic fatty . Insulin initiated: a) transferring of oleic fatty acids to lipoproteins of very low density without forming oleic lipoproteins of low density; b) highly effective oleic of fatty acids in vivo: c) becoming of biological function of locomotion. The aphysiological induction by substrate, surplus of fatty acids in food initiate negative alterations in composition of lipoproteins in opposite direction than in case of phylogenesis. When homo sapiens, herbivorous in phylogenesis, begins to misuse carnivorous (meat) food then instead of normolipoproteinemia in blood plasma under electrophoresis of lipoproteins one can initially detect transitory hyperlipoproteinemia type IV and then prolonged hyperlipoproteinemia type IIb. If patient factually passes on to carnivorous diet then hyperlipoproteinemia type V is developing. If content of exogenous fatty in food surpasses physiological capacities of its transferring in oleic triglycerides as palmitoyl-oleyl-palmitate glycerol, triglycerides as oleyl-palmitoyl-palmitate glycerol begin to form and epigenetically aphysiological non-ligand lipoproteins of very low density → lipoproteins of low density are formed. Their circulation in blood is a cause of hypertriglyceridemia, higher level of cholesterol-lipoproteins of low density, compensatory increasing of apoC-III. Then occurs induced by substrate formation of hyperlipoproteinemia initially of type IV, then of type IIb and fnally of type V. The pathogenesis of atherosclerosis and atheromotosis is activated when homo sapiens, herbivorous in phylogenesis, begin to misuse carnivorous food affecting biological functions of trophology, reaction of exotrophy (external nutrition), function of homeostasis, endoecology and function of adaptation. The formation of if fatty acids instead of oleic one is a cause of chronic defciency of energy and ATP synthesis in vivo. Insulin activates absorption of glucose by cells with purpose to use it for synthesis of oleic fatty acids. In the frst place, insulin regulates in vivo of fatty acids and only in second place of glucose.

Keyword: fat metabolism

Flavone glycosides from Sicyos angulatus and their inhibitory effects on hepatic accumulation.

A library of extracted natural materials (Korea Bioactive Natural Material Bank) have been screened to discover candidates for the treatment of non-alcoholic liver disease (NAFLD), and the 70% ethanol extract of Sicyos angulatus was found to inhibit hepatic accumulation. Bioassay-guided fractionation of this bioactive extract yielded five previously undescribed flavonoid glycosides and one previously undescribed flavonolignan glycoside along with seven known flavonoid glycosides. The chemical structures of these compounds were elucidated by a combination of extensive spectroscopic analysis, including MS, NMR and UV techniques. Eight compounds of all isolated compounds showed inhibitory effects on the accumulation induced by high concentrations of and glucose in HepG2 cells. Four selected compounds were tested for content in a dose-dependent manner (10, 20 and 40\u202fμM), and among those compounds, kaempferol 3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside showed the strongest inhibition of hepatic production in HepG2 cells. In an oil-red O staining assay, five compounds were shown to reduce hepatic accumulation better than what was observed in the vehicle control group. The present study suggests a new class of chemical entities for developing bioactive agents for the treatment of diseases caused by accumulation in the liver.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Digestibility of amino in full- canola seeds, canola meal, and canola expellers fed to broiler chickens and pigs.

Canola products including full- canola seeds (FFCS), canola meal (CM), and canola expellers (CE) have been used in diets for both broiler chickens and pigs. However, their ability to utilize the AA in canola products might be different from each other. Therefore, this study was conducted to compare the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of CP and AA in broiler chickens and growing pigs fed FFCS, CM, and CE. Three diets were prepared to contain FFCS, CM, or CE as a sole source of nitrogen. A nitrogen-free diet was prepared. In Exp. 1, a total of 272 twenty-one-day-old male broiler chickens with an initial BW of 932 ± 80.6 g were assigned to diets in a randomized complete block design with BW as a blocking factor. After 5 d of feeding, birds were euthanized by sodium pentobarbital, and ileal digesta samples were collected from distal two-third of the ileum. In Exp. 2, 16 barrows were surgically fitted with T-cannulas at the distal ileum. After 8-d recovery period, pigs (initial BW = 18.9 ± 1.17 kg) were divided into 4 blocks based on BW and assigned to a quadruplicate 4 × 2 incomplete Latin Square design with 4 diets and 2 periods. Each period consisted of 5-d adaptation and 2-d ileal digesta collection periods. Data were analyzed as a 2 × 3 factorial treatment arrangement with effects of species (broiler chickens or pigs) and diets (FFCS, CM, or CE). There were interactions (P < 0.05) between species and experimental diets in the AID of all indispensable AA except for Lys. The AID of indispensable AA in FFCS for broiler chickens was greater (P < 0.05) than for pigs. Broiler chickens also had greater (P < 0.05) AID of Arg, His, Leu, Phe, and Val in CM compared with pigs; however, there were no differences in the AID of indispensable AA in CE between broiler chickens and pigs. The basal ileal endogenous losses of CP and AA, except Trp, in pigs were greater (P < 0.05) than in broiler chickens. There were also interactions (P < 0.05) between species and experimental diets in the SID of all indispensable AA except for Lys. Broiler chickens fed the diet containing FFCS had greater (P < 0.05) SID of indispensable AA compared with pigs fed the same diet; however, the SID of indispensable AA in CM or CE were not different between broiler chickens and pigs. In conclusion, differences in digestibility of AA in canola products were affected by nonruminant animal species.

Keyword: fat metabolism

Palmitoylethanolamide Promotes a Proresolving Macrophage Phenotype and Attenuates Atherosclerotic Plaque Formation.

Objective- Palmitoylethanolamide is an endogenous fatty mediator that is synthetized from membrane phospholipids by N-acyl phosphatidylethanolamine phospholipase D. Its biological actions are primarily mediated by PPAR-α (peroxisome proliferator-activated receptors α) and the orphan receptor GPR55. Palmitoylethanolamide exerts potent anti-inflammatory actions but its physiological role and promise as a therapeutic agent in chronic arterial inflammation, such as atherosclerosis remain unexplored. Approach and Results- First, the polarization of mouse primary macrophages towards a proinflammatory phenotype was found to reduce N-acyl phosphatidylethanolamine phospholipase D expression and palmitoylethanolamide bioavailability. N-acyl phosphatidylethanolamine phospholipase D expression was progressively downregulated in the aorta of apolipoprotein E deficient (ApoE) mice during atherogenesis. N-acyl phosphatidylethanolamine phospholipase D mRNA levels were also downregulated in unstable human plaques and they positively associated with smooth muscle cell markers and negatively with macrophage markers. Second, ApoE mice were fed a high- diet for 4 or 16 weeks and treated with either vehicle or palmitoylethanolamide (3 mg/kg per day, 4 weeks) to study the effects of palmitoylethanolamide on early established and pre-established atherosclerosis. Palmitoylethanolamide treatment reduced plaque size in early atherosclerosis, whereas in pre-established atherosclerosis, palmitoylethanolamide promoted signs of plaque stability as evidenced by reduced macrophage accumulation and necrotic core size, increased collagen deposition and downregulation of M1-type macrophage markers. Mechanistically, we found that palmitoylethanolamide, by activating GPR55, increases the expression of the phagocytosis receptor MerTK (proto-oncogene tyrosine-protein kinase MER) and enhances macrophage efferocytosis, indicative of proresolving properties. Conclusions- The present study demonstrates that palmitoylethanolamide protects against atherosclerosis by promoting an anti-inflammatory and proresolving phenotype of lesional macrophages, representing a new therapeutic approach to resolve arterial inflammation.

Keyword: fat metabolism

Dietary sn-2 triacylglycerols reduced faecal lipids, calcium contents and altered in Sprague-Dawley rats.

In this study, the impact of dietary sn-2 triacylglycerol (sn-2 PTAG) on faecal lipids, calcium excretion and metabolic alternation was investigated in Sprague-Dawley (SD) rats fed with high- diet containing either palm olein (PO, sn-2 (PA) of 14.8%), sn-2 PTAG50 (sn-2\u2009PA of 56.4%) or sn-2 PTAG70 (sn-2\u2009PA of 72.4%), respectively. After 4-week feeding period, SD rats fed with sn-2 PTAGs showed reduced faecal soap fatty acids, neutral and calcium excretion compared to those of PO-fed rats, whereas a significant difference was only observed for the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Moreover, dietary sn-2 PTAG70 also showed a significant effect on decreasing serum triacylglycerol (TAG) level, reducing perirenal adipocyte size and regulating in small intestine and perirenal adipose tissue of SD rats. Significantly increased mRNA levels of genes involved in intestinal anabolism as well as catabolism were both observed in the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Meanwhile, dietary sn-2 PTAG70 also significantly up-regulated lipolysis, mitochondrial fatty oxidation and thermogenesis-related gene and protein levels in perirenal adipose tissue, which might be correlated with the reduced perirenal adipocyte size. Taken together, our findings indicated that sn-2 PTAG70 may have some beneficial effects on intestinal utilisation and metabolic activity for energy supply in visceral adipose tissue.

Keyword: fat metabolism

Probing the Global Cellular Responses to Lipotoxicity Caused by Saturated Fatty Acids.

Excessive levels of saturated fatty acids are toxic to cells, although the basis for this lipotoxicity remains incompletely understood. Here, we analyzed the transcriptome, lipidome, and genetic interactions of human leukemia cells exposed to palmitate. Palmitate treatment increased saturated glycerolipids, accompanied by a transcriptional stress response, including upregulation of the endoplasmic reticulum (ER) stress response. A comprehensive genome-wide short hairpin RNA (shRNA) screen identified >350 genes modulating lipotoxicity. Among previously unknown genetic modifiers of lipotoxicity, depletion of RNF213, a putative ubiquitin ligase mutated in Moyamoya vascular disease, protected cells from lipotoxicity. On a broader level, integration of our comprehensive datasets revealed that changes in di-saturated glycerolipids, but not other classes, are central to lipotoxicity in this model. Consistent with this, inhibition of ER-localized glycerol-3-phosphate acyltransferase activity protected from all aspects of lipotoxicity. Identification of genes modulating the response to saturated fatty acids may reveal novel therapeutic strategies for treating metabolic diseases linked to lipotoxicity.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The effect of oak tannin (Quercus robur) and hops (Humulus lupulus) on dietary nitrogen efficiency, methane emission, and milk fatty composition of dairy cows fed a low-protein diet including linseed.

The objective of this study was to test the effects of inclusion of hop pellets (HP) and oak tannin extracts (OT) alone or in combination on N efficiency, methane (CH) emission, and milk production and composition in 2 experiments with dairy cows fed low-N rations supplemented with linseed. In both experiments, 6 lactating Holstein cows were assigned to 3 dietary treatments in a 3 × 3 duplicated Latin square design (21-d periods). Cows were fed a total mixed ration at a restricted level to meet their nutrient requirements. In experiment 1, 169 g dry matter (DM) of OT or 56 g DM of HP was included separately in the control diet (C1). In experiment 2, the additives were included together (OT-HP) in the control diet (C2) similar to C1. Diet C2 was compared with a control without linseed (C0). In experiment 1, the supplementation of the control diet with OT decreased urinary N excretion by 12%. In experiment 2, the combination of OT and HP decreased urinary N by 7%. Oak tannin extracts and HP alone or in combination did not influence the daily enteric CH production of cows. Cows fed diet C0 produced 17% more enteric CH daily than those fed diet C2. Intake of diet C2, which contained 6.7% extruded linseed on a DM basis (experiment 2), decreased the sum of 6:0 to 14:0 fatty acids (-16%) and (-26%) and increased the stearic (+50%), oleic (+36%), vaccenic (trans-11 18:1; +285%), rumenic (cis-9,trans-11 18:2; +235%), and α-linolenic (+100%) in milk . The supplementation of diet C2 with the OT-HP mixture further improved the milk\'s fatty composition. Intake of the OT alone increased α-linolenic by 17.7% (experiment 1). The results of this study show that at the economically acceptable dose we tested, hops had no effect on urinary N excretion, CH emission, milk production, and milk composition. By contrast, supplementation of diets with oak tannin extract can be considered for reducing urinary N excretion. The combination of oak tannin and hops had no more effect than oak tannin alone except on the milk fatty profile, which was favorably influenced from a nutritional point of view.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Metabolic dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the energy source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive oxygen species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different metabolic conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: fat metabolism

Yeast Smell Like What They Eat: Analysis of Volatile Organic Compounds of in Growth Media Supplemented with Different Lipids.

is part of the human skin microbiota. Its volatile organic compounds (VOCs) possibly contribute to the characteristic odour in humans, as well as to microbiota interaction. The aim of this study was to investigate how the composition of the liquid medium influences the production of VOCs. Growth was performed in four media: (1) mDixon, (2) oleic (OA), (3) oleic + (OA+PA), and (4) (PA). The profiles of the VOCs were characterized by HS-SPME/GC-MS in the exponential and stationary phases. A total number of 61 VOCs was found in , among which alkanes, alcohols, ketones, and furanic compounds were the most abundant. Some compounds previously reported for (γ-dodecalactone, 3-methylbutan-1-ol, and hexan-1-ol) were also found. Through our experiments, using univariate and multivariate unsupervised (Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA)) and supervised (Projection to Latent Structures Discriminant Analysis (PLS-DA)) statistical techniques, we have proven that each tested growth medium stimulates the production of a different volatiles profile in . Carbon dioxide, hexan-1-ol, pentyl acetate, isomer5 of methyldecane, dimethyl sulphide, undec-5-ene, isomer2 of methylundecane, isomer1 of methyldecane, and 2-methyltetrahydrofuran were established as differentiating compounds among treatments by all the techniques. The significance of our findings deserves future research to investigate if certain volatile profiles could be related to the beneficial or pathogenic role of this yeast.

Keyword: fat metabolism

Changes of the Fatty Profile in Erythrocyte Membranes of Patients following 6-Month Dietary Intervention Aimed at the Regression of Nonalcoholic Fatty Liver Disease (NAFLD).

Nonalcoholic fatty liver disease (NAFLD) is closely related to the disorders of fatty acids. The pathogenesis of the disease includes an increased concentration of FFA in blood, an increase in the biosynthesis of fatty acids, and disorders in the process of -oxidation.The aim of the study was to analyze the fatty acids in erythrocyte membranes among 55 patients with NAFLD who were subjected to a 6-month dietary intervention in order to reduce fatty liver.Basic anthropometric and biochemical measurements were performed. The profile of fatty acids was measured in the membranes of erythrocytes and analyzed by gas chromatography. The dietary compliance was evaluated using 72-diary questionnaires, anthropometric measurements.With the reduction of fatty liver (p<0.01), the patients\' biochemical and anthropometric parameters were significantly improved. A significant decrease in the concentration of alanine aminotransferase (p<0.01) and asparagine aminotransferase (p<0.01) was observed, along with a decrease in the amount of insulin (p<0.05) and insulin resistance (p<0.05). Significant changes in terms of the fatty profile were observed among patients who followed the dietary intervention. There was a noticeable tendency in terms of the reduction (p<0.055) and a significant reduction of stearic (p<0.05). Significant changes in the profile of fatty acids were also associated with the reductionof palmitoleic (p<0.05) and oleic acids (p<0.05). Another statistically significant change observed was the increase in polyunsaturated fatty acids. In particular (p<0.01) the rise of eicosapentaenoic (p<0.055) and docosahexaenoic acids (p<0.55) was noted.The profile of fatty acids turned out to be a potential biomarker of the liver changes during NAFLD regression. Further research is needed to fully elucidate the usefulness and applicability of our findings in the management of NAFLD.

Keyword: fat metabolism

11β-hydroxysteroid dehydrogenase type 1 inhibitor attenuates high- diet induced cardiomyopathy.

High- diet (HFD) induces cardiac hypertrophy; however, the underlying cellular and molecular mechanisms are yet unclear. In the present study, we investigated the roles of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), an amplifier of local glucocorticoid activity, in the pathogenesis of cardiac dysfunction.Male Wistar rats were fed normal chow diet (NC) or HFD and examined the cardiac remolding and functional alteration by echocardiography and histology. Primary neonatal rat ventricular cardiomyocytes (NRCMs) treated with (PA) or infected by lentivirus were used for identifying the role by 11β-HSD1 in cardiac hypertrophy. Genome microarray of NRCMs was performed to further reveal the mechanism underlying cardiac dysfunction. induced hypertrophy in NRCMs that upregulated 11β-HSD1 expression in cardiomyocytes, which led to a significant enlargement in the cell size and expression of cardiac hypertrophy-specific genes. Conversely, a remarkable decrease in cardiomyocytes size was detected in either BVT.2733 (a selective inhibitor of 11β-HSD1)-treated or 11β-HSD1-deficient NRCMs. Furthermore, both glucocorticoid receptor (GR) antagonist RU486 and mineralocorticoid receptor (MR) antagonist spironolactone markedly attenuated the 11β-HSD1-induced cardiomyocytes hypertrophy. Genome microarray revealed that cAMP and calcium signaling pathways are potential downstream signaling pathways regulated by 11β-HSD1 in cardiomyocytes hypertrophy. Similar to in vitro results, BVT.2733 strikingly attenuated cardiac hypertrophy and improved cardiac function in HFD-fed rats.11β-HSD1 acts as an important regulator that controls the cardiac remolding via both GR and MR and the pharmacological inhibition of 11β-HSD1 could be a new therapeutic approach in preventing HFD-induced cardiac hypertrophy.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Comprehensive characterization of nanostructured carriers using laboratory and synchrotron X-ray scattering and diffraction.

The development of nanoparticles requires knowledge on the crystalline structure, polymorphic transitions and -drug interactions. This study aimed at introducing advanced techniques to characterize nanostructured carriers (NLC) comprising , oleic , stabilizer and Domperidone. Crystallinity of single components and mixtures was investigated by laboratory Small Angle X-ray Scattering (SAXS). NLC were studied with laboratory Small and Wide Angle X-ray Scattering (SWAXS). Photon Correlation Spectroscopy and Freeze Fracture Transmission Electron Microscopy were used to monitor particle size, zeta potential and shape. Stability of NLC was investigated using synchrotron X-ray Diffraction (XRD) and SAXS and laboratory SAXS. showed a lamellar structure (polymorph C), which was still present after particle preparation. Spherical 300\u202fnm-sized particles with zeta potential values above -30\u202fmV were obtained and Domperidone was incorporated in its amorphous form. During storage, no differences in synchrotron XRD spectra were seen. However, laboratory SAXS measurements showed a second lamellar structure, identified as polymorph B. Synchrotron SAXS temperature scans confirmed that polymorph B did not affect the morphology of the encapsulated drug or the shape of NLC. These results highlight the unique capabilities of laboratory and synchrotron X-ray Scattering and Diffraction for improved structural characterization of nanoparticles.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy . However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high- diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: fat metabolism

Relationship between selenium removal efficiency and production of and hydrogen by Chlorella vulgaris.

In our previous studies, Chlorella vulgaris had proven highly efficient in removing selenium (Se) from water, while the disposal of Se containing in algal biomass was still an issue of concern. Firstly, this research suggests algal Se could be released back to water, posing risks to aquatic wildlife. Thus, we further explored the possibility of using C.\xa0vulgaris to remove Se and produce and hydrogen simultaneously. Our results show the higher percentage of saturated fatty acids, especially , was found in the sulfur (S) deprived algae exposed to either selenate or selenite, although the highest content (21.9%) was found in the selenite treated algae in full BG11 medium. In addition, compared with the Se free algae, hydrogen production rate was 2.1- and 4.3-fold higher for the selenate and selenite treated algae, respectively. Se removal efficiency achieved by the selenite treated algae through accumulation and volatilization was 2.3-fold higher than the selenate treatment under hypoxic condition with S deprived, which is in contrast to the results obtained under aerobic conditions.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Mangiferin Improved Palmitate-Induced-Insulin Resistance by Promoting Free Fatty in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved Insulin Resistance.

Elevated free fatty (FFA) is a key risk factor for insulin resistance (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high- diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA in HepG2 and C2C12 cells. The model was used to quantify PA-induced accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased insulin-stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: fat metabolism

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total content was 64% of the dry cell weight and the yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: fat metabolism

Palm Oil and Beta-palmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition.

Palm oil (PO) is used in infant formulas in order to achieve (PA) levels similar to those in human milk. PA in PO is esterified predominantly at the SN-1,3 position of triacylglycerol (TAG), and infant formulas are now available in which a greater proportion of PA is in the SN-2 position (typical configuration in human milk). As there are some concerns about the use of PO, we aimed to review literature on health effects of PO and SN-2-palmitate in infant formulas.PubMed and Cochrane Database of Systematic Reviews were systematically searched for relevant studies on possible beneficial effects or harms of either PO or SN-2-palmitate in infant formula on various health outcomes.We identified 12 relevant studies using PO and 21 studies using SN-2-palmitate. Published studies have variable methodology, subject characteristics, and some are underpowered for the key outcomes. PO is associated with harder stools and SN-2-palmitate use may lead to softer stool consistency. Bone effects seem to be short-lasting. For some outcomes (infant colic, faecal microbiota, ), the number of studies is very limited and summary evidence inconclusive. Growth of infants is not influenced. There are no studies published on the effect on markers of later diseases.There is insufficient evidence to suggest that PO should be avoided as a source of in infant formulas for health reasons. Inclusion of high SN-2-palmitate blend in infant formulas may have short-term effects on stool consistency but cannot be considered essential.

Keyword: fat metabolism

Feeding high-oleic peanuts to layer hens enhances egg yolk color and oleic fatty content in shell eggs.

Previous studies have identified normal-oleic peanuts as a suitable and economical broiler feed ingredient. However, no studies to date have examined the use of high-oleic (HO) peanut cultivars as a feed ingredient for laying hens and determined the impact of feeding HO peanuts on performance and egg nutritive qualities. This project aimed to examine the use of HO peanuts as a feed ingredient for layer hens to determine the effect on performance, egg chemistry, and quality of the eggs produced. Forty-eight 40-wk-old layer hens were fed a conventional soybean meal + corn control diet or a HO peanut + corn diet for 10 wk in conventional battery cages. Body and feed weights were collected weekly. Pooled egg samples were analyzed for quality, analysis, and peanut protein allergenicity. There were no significant differences in hen performance or egg quality as measured by USDA grade quality, egg albumen height, or egg Haugh unit between the treatment groups. However, eggs produced from layer hens fed the HO peanut + corn diet had reduced egg weights relative to the controls (P = 0.0001). Eggs produced from layer hens fed the HO peanut diet had greater yolk color scores (P < 0.0001), HO fatty (P < 0.0001), and β-carotene (P < 0.0001) levels in comparison to the controls. Eggs produced from hens fed the control diet had greater and stearic saturated fatty acids (P < 0.0001), and trans (P < 0.0001) content compared to eggs produced from hens fed the HO peanut diet. All egg protein extracts from all treatments at each time point were non-reactive with rabbit anti-peanut agglutinin antibodies. This study identifies HO peanuts as an abundant commodity that could be used to support local agricultural markets of peanuts and poultry within the southeastern United States and be of economic advantage to producers while providing a potential health benefit to the consumer with improved egg nutrition.Published by Oxford University Press on behalf of Poultry Science Association 2018.

Keyword: fat metabolism

Comparison of monoculture and mixed culture (Scenedesmus obliquus and wild algae) for C, N, and P removal and production.

This study compared the efficiency of nutrient removal and accumulation by a monoculture of Scenedesmus obliquus and mixed cultures of microalgae. The highest removal efficiencies of ammonium (99.2%), phosphate (91.2%), and total organic carbon (83.6%) occurred in the monoculture. All the mixed cultures were dominated by S. quadricauda; in some mixed cultures, the proportions of Chlamydomonas reinhardtii and C. microsphaera reached >\u200920%. The content and production in the monoculture were 15.9% and 52.3\xa0mg\xa0kg, respectively, significantly higher than those in all the mixed cultures of microalgae. In all the mixed cultures, the proportion of was >\u200950%. The results suggest that the monoculture had advantages over the mixed culture of microalgae in terms of nutrient removal and production.

Keyword: fat metabolism

Upregulation of miR-181a impairs by targeting PPARα expression in nonalcoholic fatty liver disease.

Recent studies have reported elevated expression of miR-181a in patients with non-alcoholic fatty liver disease (NAFLD), suggesting that it may play an important role in liver and insulin resistance. We aimed to investigate the effect of miR-181a in and find new treatments for NAFLD. The expression level of miR-181a in NAFLD patient serum and a (PA)-induced NAFLD cell model was examined by Q-PCR. Oil red O staining and triglyceride assays were used to assess accumulation in hepatocytes. Western blotting was used to detect the protein expression levels of peroxisome proliferator-activated receptor-α (PPARα) and the fatty β-oxidation-related genes. Direct interactions were validated by dual-luciferase reporter gene assays. MiR-181a expression was significantly upregulated in the serum of NAFLD patients and PA-induced hepatocytes. Inhibition of miR-181a expression resulted in the increased expression of PPARα and its downstream genes, and PA-induced accumulation in hepatocytes was also inhibited. Upregulation of miR-181a resulted in the downregulation of its direct target PPARα and downstream gene expression of PPARα as well as aggravated accumulation in hepatocytes. At the same time, the increased expression of PPARα can offset accumulation in hepatocytes induced by miR-181a mimics. This study demonstrates that reducing the expression of miR-181a may improve in NAFLD. The downregulation of miR-181a expression can be a therapeutic strategy for NAFLD by modulating its target PPARα.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Early and reversible changes to the hippocampal proteome in mice on a high- diet.

The rise in global obesity makes it crucial to understand how diet drives obesity-related health conditions, such as premature cognitive decline and Alzheimer\'s disease (AD). In AD hippocampal-dependent episodic memory is one of the first types of memory to be impaired. Previous studies have shown that in mice fed a high- diet (HFD) episodic memory is rapidly but reversibly impaired.In this study we use hippocampal proteomics to investigate the effects of HFD in the hippocampus. Mice were fed either a low- diet (LFD) or HFD containing either 10% or 60% (Kcal) from for 3\u2009days, 1\u2009week or 2\u2009weeks. One group of mice were fed the HFD for 1\u2009week and then returned to the LFD for a further week. Primary hippocampal cultures were challenged with (PA), the most common long-chain saturated FA in the Western diet, and with the anti-inflammatory, n-3 polyunsaturated FA, docosahexaenoic (DHA), or a combination of the two to ascertain effects of these fatty acids on dendritic structure.HFD-induced changes occur in hippocampal proteins involved in , inflammation, cell stress, cell signalling, and the cytoskeleton after 3\u2009days, 1\u2009week and 2\u2009weeks of HFD. Replacement of the HFD after 1\u2009week by a low- diet (LFD) for a further week resulted in partial recovery of the hippocampal proteome. Microtubule-associated protein 2 (MAP2), one of the earliest proteins changed, was used to investigate the impact of fatty acids (FAs) on hippocampal neuronal morphology. PA challenge resulted in shorter and less arborised dendrites while DHA had no effect when applied alone but counteracted the effects of PA when FAs were used in combination. Dendritic morphology recovered when PA was removed from the cell culture media.This study provides evidence for the rapid and reversible effects of diet on the hippocampal proteome and the impact of PA and DHA on dendritic structure.

Keyword: fat metabolism

Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic fatty liver disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high- diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of fatty liver disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Dietary habits affect fatty composition of visceral adipose tissue in subjects with colorectal cancer or obesity.

Aim of this study was to identify a possible relationship among dietary fatty acids (FA) intake, FA adipose tissue (AT) profile and cancer condition in lean vs obese subjects affected or not by colorectal cancer (CRC). Actually, inadequate dietary habits together with physical inactivity are primary determinants of obesity and cancer risk. Changes in play a crucial role in different types of cancer and key enzymes involved in -metabolic pathways, such as stearoyl-coA-desaturase 1 (SCD-1), are differentially expressed in normal and cancer tissues.Food frequency questionnaires (FFQ) were analyzed by Winfood software. FA were assessed by gas-liquid chromatography in visceral AT samples. Estimated desaturase activities were calculated as precursor FA/product FA ratio. Desaturase gene expressions were evaluated by RT-qPCR.Lean and obese CRC subjects showed inadequate dietary habits. In particular, lean CRC subjects showed increase in the intake of saturated FA, specifically (p\u2009=\u20090.0042) and stearic (p\u2009=\u20090.0091), and a corresponding reduction of monounsaturated FA consumption, in particular oleic (p\u2009=\u20090.002) with respect to lean without CRC. Estimated SCD-1 activity in AT was increased in all the groups vs lean without CRC (pANOVA\u2009=\u20090.029).Unhealthy eating habits, characterizing obese and CRC subjects, may influence the visceral AT profile and contribute to the alteration of the metabolic pathways. The quality of the diet, other than the quantity, can have a main role in the establishment of inflammatory microenvironment and in metabolic changes favouring CRC.

Keyword: fat metabolism

9-PAHSA promotes browning of white via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract obesity in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white . 9-PAHSA, an endogenous mammalian , which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of obesity.Copyright © 2018. Published by Elsevier Inc.

Keyword: fat metabolism

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high- diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and . We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: fat metabolism

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on -induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose , such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in , including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: fat metabolism

Telmisartan protects against high glucose/high -induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high -induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the of fatty acids (FA) and only in the second turn in glucose ; regulation of FA in cells started millions of years earlier than that of glucose . Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: fat metabolism

Resveratrol Ameliorates Droplet Accumulation in Liver Through a SIRT1/ ATF6-Dependent Mechanism.

droplets (LDs) are dynamic organelles that store neutral lipids during times of energy excess, and an increased accumulation of LDs in the liver is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high- diet (HFD) and were used to induce hepatic steatosis in mouse liver and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in liver and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

Relationship between acyl- and sterol metabolisms in diatoms.

Diatoms are a phylum of unicellular photosynthetic eukaryotes living in oceans and fresh waters, characterized by the complexity of their plastid, resulting from a secondary endosymbiosis event. In the model diatom Phaeodactylum tricornutum, fatty acids (FAs) are synthesized from acetyl-CoA in the stroma of the plastid, producing . FAs are elongated and desaturated to form very-long chain polyunsaturated fatty acids (VLC-PUFAs) in domains of the endomembrane system that need to be identified. Synthesis of VLC-PUFAs is coupled with their import to the core of the plastid via the so-called "omega" pathway. The biosynthesis of sterols in diatoms is likely to be localized in the endoplasmic reticulum as well as using precursors deriving from the mevalonate pathway, using acetyl-CoA as initial substrate. These metabolic modules can be characterized functionally by genetic analyzes or chemical treatments with appropriate inhibitors. Some \'metabolic modules\' are characterized by a very low level of metabolic intermediates. Since some chemical treatments or genetic perturbation of induce the accumulation of these intermediates, channeling processes are possibly involved, suggesting that protein-protein interactions might occur between enzymes within large size complexes or metabolons. At the junction of these modules, metabolic intermediates might therefore play dramatic roles in directing carbon fluxes from one direction to another. Here, acetyl-CoA seems determinant in the balance between TAGs and sterols. Future lines of research and potential utilization for biotechnological applications are discussed.Published by Elsevier B.V.

Keyword: fat metabolism

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and .

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high- diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fat metabolism

Factors associated with the content of mammary-synthesized fatty acids in milk : A meta-analysis.

Consumption of specific fatty acids (FA) that are synthesized in the mammary gland, namely de novo FA, has implications for human health. The objective of the present meta-analysis was to study the associations between milk content of de novo FA, with (1) diet composition, and (2) milk production and composition. Milk FA data from 96 peer-reviewed studies published between 1990 and 2016 that included 324 treatment means from 83 bovine experiments, 36 treatment means from 12 caprine experiments, and 40 treatment means from 12 ovine experiments were used in this analysis. Individual species models including the fixed effect of experiment were fitted using multiple regression to explain milk content of de novo FA as a function of diet composition and milk production and composition variables. We also evaluated replacing the effect of the experiment by the effect of the experiment nested in the laboratory at which the research had been conducted, and the effect of the laboratory. Butyric content in milk was positively but weakly related to dietary ether extract in does and ewes. Lauric, myristic, and contents in milk were negatively related to dietary ether extract in does and to a somewhat lesser extent in cows and ewes. The results confirm that the inclusion of lipids in the diet may not only affect the availability of preformed FA but also the profile of FA synthesized de novo in the mammary gland. Most of the variation in all prediction models was explained by the experiment or by the laboratory if the latter was included in the model. The ample variation in analytical methods reported by the different research groups suggests that differences in analytical protocols might explain a substantial proportion of the variation in de novo FA profile. A main conclusion of this study is the potential influence of differences in analytical procedures to explain the variation in de novo FA profile. Standardization of methods of FA analysis to improve reproducibility seems to be an aspect of importance to this area of research.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Flaxseed oil ameliorated high--diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts.

α-Linolenic (ALA) is a plant-derived omega-3 unsaturated fatty that is rich in flaxseed oil (FO). The effect of FO on bone health is controversial. This study aims to evaluate the effect of FO on bone damage induced by a high- diet (HFD) and to explore the possible mechanism.Male Sprague-Dawley rats were fed a normal control diet (NC, 10% ), FO diet (NY, 10% ), HFD (60% ), or HFD containing 10% FO (HY, 60% ) for 22\u2009weeks. Micro CT and three-point bending tests were conducted to evaluate bone microstructure and biomechanics. Serum was collected for the detection of ALP, P1NP, and CTX-1. Rat primary osteoblasts (OBs) were treated with different concentrations of ALA with or without (PA) treatment. The ALP activity, osteogenic-related gene and protein expression were measured.Rats in the HFD group displayed decreased biomechanical properties, such as maximum load, maximum fracture load, ultimate tensile strength, stiffness, energy absorption, and elastic modulus, compared with the NC group ( \xa0<\u20090.05). However, HY attenuated the HFD-induced decreases in bone biomechanical properties, including maximum load, maximum fracture load, and ultimate tensile strength (\u2009<\u20090.05). Trabecular bone markers such as trabecular volume bone mineral density (Tb. vBMD trabecular bone volume/total volume (Tb. BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) were decreased, trabecular separation (Tb. Sp) and the structure model index (SMI) were increased in the HFD group compared with the NC group, and all parameters were remarkably improved in the HY group compared to the HFD group (p\u2009<\u20090.05). However, cortical bone markers such as cortical volume bone mineral density (Ct. vBMD), cortical bone volume/total volume (Ct. BV/TV) and cortical bone thickness (Ct. Th) were not significantly different among all groups. Moreover, the serum bone formation markers ALP and P1NP were higher and the bone resorption marker CTX-1 was lower in the HY group compared with levels in the HFD group. Compared with the NC group, the NY group had no difference in the above indicators. In rat primary OBs, PA treatment significantly decreased ALP activity and osteogenic gene and protein (β-catenin, RUNX2, and osterix) expression, and ALA dose-dependently restored the inhibition induced by PA.FO might be a potential therapeutic agent for HFD-induced bone loss, most likely by promoting osteogenesis.© The Author(s). 2019.

Keyword: fat metabolism

Anthocyanins ameliorate palmitate-induced inflammation and insulin resistance in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade inflammation leading to insulin resistance. Adipocyte differentiation inhibitors are expected to be effective in preventing obesity and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from metabolic stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, inflammation, and insulin resistance in 3T3-L1 adipocytes. ACN extract pretreatment reduces accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces inflammation with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces insulin resistance by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce insulin pathway with levels higher than insulin control cells, supporting an insulin sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against obesity comorbidities, due to their protective effects against inflammation/insulin resistance in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and insulin resistance conditions related to obesity.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

1,25-Dihydroxyvitamin D regulates and glucose utilization in differentiated 3T3-L1 adipocytes.

It is well established that adipose tissue can both store and metabolize vitamin D. The active form of vitamin D, 1,25 dihydroxyvitamin D [1,25(OH)D], regulates adipocyte differentiation and inflammation, highlighting the multifaceted role that vitamin D plays in adipose tissue physiology. However, there is limited evidence regarding vitamin D regulation of mature adipocyte . We hypothesize that 1,25(OH)D alters and glucose in differentiated 3T3-L1 adipocytes to reduce triacylglycerol (TAG) accumulation. In this study, 1,25(OH)D (10\u202fnmol/L) stimulated a 21% reduction in TAG accumulation in differentiated 3T3-L1 adipocytes after 4\u202fdays (P\u202f=\u202f.01) despite a significant increase in fatty uptake (P\u202f<\u202f.01). Additionally, 1,25(OH)D stimulated a 2.5-fold increase in CO production from [1-C] (P\u202f<\u202f.01), indicative of an elevated rate of fatty β-oxidation, while stimulating a 9% reduction in de novo fatty synthesis (P\u202f=\u202f.03). Interestingly, d-[U-C]glucose incorporation into fatty acids was reduced by 30% in response to 1,25(OH)D (P\u202f<\u202f.01), indicating a reduced contribution of glucose as a substrate for de novo lipogenesis. Consistent with these findings, mRNA expression of the anaplerotic enzyme pyruvate carboxylase was reduced by 41% (P\u202f<\u202f.01). In summary, 1,25(OH)D stimulated fatty oxidation and reduced TAG accumulation in differentiated adipocytes. Furthermore, 1,25(OH)D reduced glucose utilization as a substrate for fatty synthesis potentially by downregulating pyruvate carboxylase and stimulating glucose disposal as glycerol. Collectively, these 1,25(OH)D-induced changes in and glucose utilization may contribute to the reduction in TAG accumulation and be protective against excessive mass accumulation and associated metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Neuroprotective effects of vitamin D on high diet- and -induced enteric neuronal loss in mice.

The role of vitamin D in obesity and diabetes is debated. Obese and/or diabetic patients have elevated levels of free fatty acids, increased susceptibility to gastrointestinal symptoms and are suggested to have altered vitamin D balance. The enteric nervous system is pivotal in regulating gastrointestinal activity and high diet (HFD) has been shown to cause loss of enteric neurons in ileum and colon. This study investigates the effect of vitamin D on HFD- and -induced enteric neuronal loss in vivo and in vitro.Mice were fed either a normal diet (ND) or HFD supplemented with varying levels of vitamin D (from 0x to 20x normal vitamin D level) for 19 weeks. Ileum and colon were analyzed for neuronal numbers and remodeling. Primary cultures of myenteric neurons from mouse small intestine were treated with (4x10M) and/or 1α,25-hydroxy-vitamin D3 (VD, 10- 10M) with or without modulators of and VD pathways. Cultures were analyzed by immunocyto- and histochemical methods.Vitamin D supplementation had no effect on enteric neuronal survival in the ND group. HFD caused substantial loss of myenteric neurons in ileum and colon. Vitamin D supplementation between 0-2x normal had no effect on HFD-induced neuronal loss. Supplementation with 20x normal, prevented the HFD-induced neuronal loss. In vitro supplementation of VD prevented the -induced neuronal loss. The VD receptor (VDR) was not identified in enteric neurons. Enteric glia expressed the alternative VD receptor, protein disulphide isomerase family A member 3 (PDIA3), but PDIA3 was not found to mediate the VD response in vitro. Inhibition of peroxisome proliferator-activated receptor gamma (PPARγ) and immune neutralization of isocitrate lyase prevented the VD mediated neuroprotection to exposure.Results show that VD protect enteric neurons against HFD and induced neuronal loss. The mechanism behind is suggested to be through activation of PPARγ leading to improved neuronal peroxisome function and of neuronal intermediates.

Keyword: fat metabolism

Protocatechuic -Ameliorated Endothelial Oxidative Stress through Regulating Acetylation Level via CD36/AMPK Pathway.

As one of the main metabolites of anthocyanin, protocatechuic (PCA) possesses strong antioxidant activity. In the present study, we explored the capacity of PCA on the alleviation of endothelial oxidative stress and investigated the underlying mechanisms using RNA sequencing (RNA-Seq). In comparison with (PA)-treated cells, PCA (100 μM) significantly decreased the generations of 3-nitrotyrosine (3-NT) and 8-hydroxydeoxyguanosine (8-OHdG) (0.82 ± 0.01 vs 1.16 ± 0.05 and 0.80 ± 0.01 vs 1.48 ± 0.15, respectively, p < 0.01), two biomarkers of oxidative damage, and restored the levels of nitric oxide (NO) (0.97 ± 0.04 vs 0.54 ± 0.02, p < 0.01) and mitochondrial membrane potential (MMP) (0.96 ± 0.03 vs 0.86 ± 0.02, p < 0.01) in human umbilical vein endothelial cells (HUVECs). PCA also obviously reduced the level of reactive oxygen species (ROS) (0.86 ± 0.15 vs 2.67 ± 0.09, p < 0.01) in aorta from high- diet (HFD)-fed mice. RNA-Seq and Western blot analysis indicated that PCA markedly reduced the expression of cluster of differentiation 36 (CD36), a membrane fatty transporter, and reduced the generations of adenosine triphosphate (ATP) and acetyl coenzyme A (Ac-CoA). These effects of PCA were associated with decreased level of acetylated-lysine and restored the activity of manganese-dependent superoxide dismutase (MnSOD) through reducing the generation of Ac-CoA or activating Sirt1 and Sirt3 via a CD36/AMP-kinase (AMPK) dependent pathway.

Keyword: fat metabolism

Concomitant production of chitosan and lipids from a newly isolated Mucor circinelloides ZSKP for biodiesel production.

A newly-isolated oleaginous fungus Mucor circinelloides ZSKP concurrently yielded 21.4% lipids and 11.2% chitosan per gram of biomass. Parameters affecting the co-production were identified using Plackett-Burman design and were statistically optimized using Response Surface Methodology, which resulted in a 3-fold improvement in production. The profile showed a high content of unsaturated fatty acids including oleic (37%), linolenic (14%) and linoleic acids (19%), while was the major saturated fatty (21%). A comparative study to evaluate the efficacy of enzymatic (lipase) and chemical treatments for biodiesel production from fungal lipids and sunflower oil revealed enhanced production of biodiesel from fungal lipids. Synthesized biodiesel from M. circinelloides ZSKP satisfied standard specifications and had a higher cetane number (56), lower kinematic viscosity (4.6\u202fmm/s) and lower number (0.03) compared to sunflower oil. Results suggest Mucor circinelloides ZSKP is a promising candidate for implementation of the biorefinery concept.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Nutrient-Dependent Changes of Protein Palmitoylation: Impact on Nuclear Enzymes and Regulation of Gene Expression.

Diet is the main environmental stimulus chronically impinging on the organism throughout the entire life. Nutrients impact cells via a plethora of mechanisms including the regulation of both protein post-translational modifications and gene expression. Palmitoylation is the most-studied protein lipidation, which consists of the attachment of a molecule of to residues of proteins. -palmitoylation is a reversible cysteine modification finely regulated by palmitoyl-transferases and acyl-thioesterases that is involved in the regulation of protein trafficking and activity. Recently, several studies have demonstrated that diet-dependent molecules such as insulin and fatty acids may affect protein palmitoylation. Here, we examine the role of protein palmitoylation on the regulation of gene expression focusing on the impact of this modification on the activity of chromatin remodeler enzymes, transcription factors, and nuclear proteins. We also discuss how this physiological phenomenon may represent a pivotal mechanism underlying the impact of diet and nutrient-dependent signals on human diseases.

Keyword: fat metabolism

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for obesity and diabetes.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fat metabolism

Effect of Microalgae Polysaccharides on Biochemical and Metabolomics Pathways Related to Plant Defense in Solanum lycopersicum.

Microalgae are photosynthetic microorganisms that produce several bioactive molecules that have received considerable attention in scientific and industrial communities. Today, many plant biostimulants including seaweed extracts and polysaccharides are used in agriculture. However, microalgae have not been largely exploited in this field as a potential source of plant bio stimulants. This study investigated the biostimulatory effects of microalgae polysaccharides on different metabolomic and biochemical pathways related to plant defense. 0.2\xa0mg\xa0mL of crude polysaccharides extracted from four green microalgae strains was injected into tomato plants (Solanum lycopersicum). β-1,3-glucanase activity, remodeling, phenylalanine ammonia lyase (PAL), Lipoxygenase (LOX), and antioxidant enzyme (APX, POD and CAT) activities were evaluated 48\xa0h after treatment. Plants treated with crude polysaccharides extracted from. C. vulgaris and C. sorokiniana exhibited a significant increase in β-1,3-glucanase activity. Accordingly, C. sorokiniana crude polysaccharides had a significant stimulatory effect on PAL activity with a percentage increase of 188.73% compared to the control. GC/MS quantitative lipidomics analysis revealed that treatment with D. salina, C. sorokiniana, and C. reinhardtii crude polysaccharides increased PUFA content by 50.37%, 34.46%, and 33.37% respectively. Microalgae polysaccharides also enhanced stearic , , and VLCFA content, the optimal value of which increased by 45.50%, 32.83%, and 60.60% respectively under treatment with C. reinhardtii crude polysaccharides compared with the control. C. vulgaris and C. reinhardtii crude polysaccharides also exhibited higher APX and POD activity respectively. The present results therefore indicate the potentiality of microalgae crude polysaccharides as a promising renewable bio resource in the development plant bio stimulants.

Keyword: fat metabolism

Saturated fatty stimulates production of extracellular vesicles by renal tubular epithelial cells.

Lipotoxicity, an accumulation of intracellular metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of metabolic disease. , a predominant derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated (PA), and BSA-conjugated oleic (OA) for 24-48\xa0h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30-150\xa0nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of -induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of -induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keyword: fat metabolism

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated -induced insulin resistance that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty , insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Sodium Orthovanadate Changes Fatty Composition and Increased Expression of Stearoyl-Coenzyme A Desaturase in THP-1 Macrophages.

Vanadium compounds are promising antidiabetic agents. In addition to regulating glucose , they also alter . Due to the clear association between diabetes and atherosclerosis, the purpose of the present study was to assess the effect of sodium orthovanadate on the amount of individual fatty acids and the expression of stearoyl-coenzyme A desaturase (SCD or Δ-desaturase), Δ-desaturase, and Δ-desaturase in macrophages. THP-1 macrophages differentiated with phorbol 12-myristate 13-acetate (PMA) were incubated in vitro for 48\xa0h with 1\xa0μM or 10\xa0μM sodium orthovanadate (NaVO). The estimation of fatty composition was performed by gas chromatography. Expressions of the genes SCD, fatty desaturase 1 (FADS1), and fatty desaturase 2 (FADS2) were tested by qRT-PCR. Sodium orthovanadate in THP-1 macrophages increased the amount of saturated fatty acids (SFA) such as and stearic , as well as monounsaturated fatty acids (MUFA)-oleic and palmitoleic . Sodium orthovanadate caused an upregulation of SCD expression. Sodium orthovanadate at the given concentrations did not affect the amount of polyunsaturated fatty acids (PUFA) such as linoleic , arachidonic , eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA). In conclusion, sodium orthovanadate changed SFA and MUFA composition in THP-1 macrophages and increased expression of SCD. Sodium orthovanadate did not affect the amount of any PUFA. This was associated with a lack of influence on the expression of FADS1 and FADS2.

Keyword: fat metabolism

Plasma fatty acids in de novo lipogenesis pathway are associated with diabetogenic indicators among adults: NHANES 2003-2004.

Insulin regulates fatty acids (FAs) in the blood; conversely, FAs may mediate insulin sensitivity and are potentially modifiable risk factors of the diabetogenic state.The objective of our study was to examine the associations between plasma concentrations of FAs, fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among individuals (n\xa0=\xa01433) in the NHANES (2003-2004).Plasma concentrations of 24 individual FAs were considered individually and in subgroups, per chemical structure. Study participants were categorized in diabetogenic groups: Group 1 (HbA1c ≥6.5% or FPG ≥126 mg/dL), Group 2 (HbA1c 5.7% to <6.5% or FPG 100 to <126 mg/dL), and Group 3 (HbA1c <5.7% and FPG <100 mg/dL). We assessed associations between diabetogenic groups and plasma FAs in multivariate multinomial regressions (with Group 3 as the reference).Overall, 7.0% of study participants were in Group 1; 33.3% were in Group 2. Plasma concentrations of several individual FAs, including even-chain saturated FAs (SFAs; myristic, , stearic acids) and monounsaturated FAs (MUFAs; cis-vaccenic, oleic acids), were respectively associated with greater odds of Groups 1 and 2 status, adjusting for covariates. Higher concentrations of SFA and MUFA subgroups (highest compared with lowest quartile) were associated with increased odds of Group 2 status [SFAs adjusted OR (aOR): 1.51 (95% CI: 1.05, 2.18); MUFAs aOR: 1.78 (95% CI: 1.11, 2.85)]. Higher eicosapentaenoic plasma concentration was associated with decreased odds of Group 1 status [quartile 4 aOR: 0.41 (95% CI: 0.17, 0.95)].Higher plasma concentrations of SFAs and MUFAs, primary de novo lipogenesis products, were associated with elevated FPG and HbA1c in a nationally representative study population in the United States. Additional studies are necessary to elucidate potential causal relationships between FAs (from endogenous production and dietary consumption) and diabetogenic indicators, as well as clinical implications for managing diabetes and prediabetes.

Keyword: fat metabolism

PKCζ Phosphorylates SIRT6 to Mediate Fatty β-Oxidation in Colon Cancer Cells.

Protein kinase C (PKC) has critical roles in regulating anabolism and catabolism. PKCζ, a member of atypical PKC family, has been reported to mediate glucose . However, whether and how PKCζ regulates tumor cells fatty β-oxidation are unknown. Here, we report that the phosphorylation of SIRT6 is significantly increased after (PA) treatment in colon cancer cells. PKCζ can physically interact with SIRT6 in vitro and in vivo, and this interaction enhances following PA treatment. Further experiments show that PKCζ is the phosphorylase of SIRT6 and phosphorylates SIRT6 at threonine 294 residue to promote SIRT6 enrichment on chromatin. In the functional study, we find that the expression of ACSL1, CPT1, CACT, and HADHB, the genes related to fatty β-oxidation, increases after PA stimulation. We further confirm that PKCζ mediates the binding of SIRT6 specifically to the promoters of fatty β-oxidation-related genes and elicits the expression of these genes through SIRT6 phosphorylation. Our findings demonstrate the mechanism of PKCζ as a new phosphorylase of SIRT6 on maintaining tumor fatty β-oxidation and define the new role of PKCζ in homeostasis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

[Lipolysis in very low density lipoproteins - locus minoris resistentiae - in the pathogenesis of hypertriglyceridemia. Positive effects of diet, polyenic fatty acids, statins and fibrates.]

Inhibition of hydrolysis of and oleic triglycedires (TG) in very low density lipoproteins (VLDL), slow formation of active apoВ-100 conformation, blockade of апоЕ/В-100 ligand formation in VLDL and their reduced uptake by insulin-dependent cells cause hypertriglyceridemia (HTG). and oleic VLDL (>80% total VLDL) are not converted in low density lipoproteins (LDL). Atherosclerosis is not an alimentary deficiency of polyenic fatty acids (PFA), but results from low in vivo bioavailability of PFA in LDL against the background of high dietary FA and LDL. Plasma PFA content and cellular PFA deficiency are as high as LDL cholesterol (CL). Primary prevention of atherosclerosis should be based on a decrease in dietary content of saturated FA, trans FA and a moderate increase in PFA. It seems highly unlikely that the xeobiotics statins, fibrates and probucol produce pleiotropic biological effects in vivo. These effects are brought about by phylogenetically early humoral mediators eicosanoids: prostacyclins, prostaglandins, thromboxanes, leukotrienes, and resolvins. It is reasonable to suggest that all preparations which act according to the same algorithm activate TG hydrolysis in VLDL and normalize cellular uptake of PFA in linoleic and linolenic LDL via apoВ-100 endocytosis. Atherosclerosis is a syndrome of cellular deficiency of essential polyenic FA.

Keyword: fat metabolism

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high- diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated accumulation and macrophage-mediated inflammation.

Keyword: fat metabolism

Rosiglitazone ameliorates -induced cytotoxicity in TM4 Sertoli cells.

The Sertoli cell is the only somatic cell within the seminiferous tubules, and is vital for testis development and spermatogenesis. Rosiglitazone (RSG) is a member of the thiazolidinedione family and is a peroxisome proliferator-activated receptor-γ (PPARγ) agonist. It has been reported that RSG protects various types of cells from fatty -induced damage. However, whether RSG serves a protective role in Sertoli cells against (PA)-induced toxicity remains to be elucidated. Therefore, the aim of the present study was to investigate the effect of RSG on PA-induced cytotoxicity in Sertoli cells. MTT assay and Oil Red O staining revealed that RSG ameliorated the PA-induced decrease in TM4 cell viability, which was accompanied by an alleviation of PA-induced accumulation in cells. In primary mouse Sertoli cells, RSG also showed similar protective effects against PA-induced lipotoxicity. Knockdown of PPARγ verified that RSG exerted its protective role in TM4 cells through a PPARγ-dependent pathway. To evaluate the mechanism underlying the protective role of RSG on PA-induced lipotoxicity, the present study analyzed the effects of RSG on PA uptake, and the expression of genes associated with both fatty oxidation and triglyceride synthesis. The results demonstrated that although RSG did not affect the endocytosis of PA, it significantly elevated the expression of carnitine palmitoyltransferase (CPT)-1A, a key enzyme involved in fatty oxidation, which indicated that the protective effect of RSG may have an important role in fatty oxidation. On the other hand, the expression of CPT1B was not affected by RSG. Moreover, the expression levels of diacylglycerol O-acyltransferase (DGAT)-1 and DGAT2, both of which encode enzymes catalyzing the synthesis of triglycerides, were not suppressed by RSG. The results indicated that RSG reduced PA-induced accumulation by promoting fatty oxidation mediated by CPT1A. The effect of RSG in protecting cells from lipotoxicity was also found to be specific to Sertoli cells and hepatocytes, and not to other cell types that do not store excess in large quantities, such as human umbilical vein endothelial cells. These findings provide insights into the cytoprotective effects of RSG on Sertoli cells and suggest that PPARγ activation may be a useful therapeutic method for the treatment of Sertoli cell dysfunction caused by dyslipidemia.

Keyword: fat metabolism

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral metabolic process" and "Regulation of storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body energy .

Keyword: fat metabolism

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high- diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and -uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA into herbivorous dry land-living species with oleic type of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: fat metabolism

Evidence for perturbed metabolic patterns in bipolar disorder subjects associated with lithium responsiveness.

Bipolar disorder (BD) is multifactorial mood disorder characterized by alternating episodes of hyperactive mania and severe depression. Lithium is one of the most preferred drug used as mood stabilizer in treating BD. In this study, we examined the changes in plasma metabolome in BD subjects in the context of lithium responsiveness. Plasma samples from clinically defined, age and gender matched unrelated healthy controls and BD subjects (lithium responders and non-responders) were obtained and processed in positive and negative mode using untargeted liquid chromatography/mass spectrometry analysis. We identified significant alterations in plasma levels of dopamine along with its precursors (tyrosine and phenylalanine), branched chain amino such as valine and excitatory neurotransmitter glutamate between healthy control and BD subjects. molecules such as, eicosenoic and retinyl ester also showed distinguished patterns between control and BD individuals. Lithium responsiveness was markedly associated with significant differences in proline, L-gamma-glutamyl-isoleucine, dopamine, methyl ester, cholesterol sulfate, androsterone sulfate and 9S,12S,13S-triHOME levels. Altered metabolites enriched with key biochemical pathways associated with neuropsychiatry disorders. We hypothesize that BD pathogenesis and lithium responsiveness is associated with impaired homeostasis of amino and .Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

Ellagic Suppresses the Oxidative Stress Induced by Dietary-Oxidized Tallow.

Dietary tallow was thermally oxidized at 180°C in an open fryer. The oxidized tallow (OT) and unoxidized tallow were characterized for oxidation parameters and fatty composition using GC-MS. Tallow samples were fed to rabbits along with 50, 100, and 150\u2009mg/kg/day of ellagic (EA) for three weeks. Results revealed that the peroxide value (PV) and thiobarbituric reactive substances (TBARS) significantly increased, while radical scavenging activity (RSA) of the tallow decreased significantly with oxidation. GC-MS analysis showed eight fatty acids in the tallow samples, where (48.5-49.7\u2009g/100\u2009g), linoleic (18.7-23.7\u2009g/100\u2009g), stearic (13.5-15.6\u2009g/100\u2009g), and margaric (6.32-6.42\u2009g/100\u2009g) were the major fatty acids. Animal studies showed that oxidized tallow (OT) alone or in combination with EA significantly altered the body weight of the rabbits. Serum biochemical parameters and renal function tests were affected by OT and ameliorated by EA. The toxic effects of OT on haematological indices were minimized by EA. The supplementation of OT alone had significant effects on the liver structure and functions. The coadministration of EA reduced the toxic properties of OT on the liver, by increasing the antioxidant (GSH) system. The rabbit heart was also affected by the OT, which was ameliorated by EA supplementation. These results suggested that the supplementation of EA was beneficial against the OT-induced oxidative stress and may be considered for foods containing oxidized lipids.

Keyword: fat metabolism

The effect of enterolactone on liver precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on fractions fatty acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free fatty acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their fatty acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of fatty acids were evaluated by Western Blot.Enterolactone modified fatty acids composition in FFA, DAG and TAG fractions. In conjunction with overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated fatty acids in each of the investigated fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Molecular mechanism of activation of the immunoregulatory amidase NAAA.

Palmitoylethanolamide is a bioactive that strongly alleviates pain and inflammation in animal models and in humans. Its signaling activity is terminated through degradation by -acylethanolamine amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand\'s acyl chain.

Keyword: fat metabolism

Rare ginsenosides ameliorate overload-induced myocardial insulin resistance via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 diabetes. However, their pharmacological studies regarding myocardial fuel and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fat metabolism

Influence of Resveratrol on Sphingolipid in Hepatocellular Carcinoma Cells in Overload State.

Obesity is characterized by increased long chain fatty acids (LCFA) uptake and impaired in hepatocytes. Consequently, an enhanced intracellular content, including sphingolipids, may lead to lipotoxicity. It is believed that resveratrol (RSV), one of the most extensively studied plant-derived polyphenols, and its interaction with sphingolipid may constitute one of the major therapeutic targets for cancer and metabolic diseases treatment.The aim of this study was to ascertain, whether resveratrol may affect sphingolipid metabolic pathways, enzymes and transporters in a overload state.The experiments were conducted on hepatocellular carcinoma cells (HepG2) incubated with RSV and/or (PA) at the concentration of 0.5 mM and 50 µM, respectively for 16h. Intra- and extracellular sphingolipid concentrations were assessed by high-performance liquid chromatography and gas liquid chromatography. Moreover, the expression of caspase 3, selected fatty transporters and sphingolipid pathway proteins were estimated by Western Blot.RSV alone and together with PA significantly increased the intracellular concentration of ceramide, sphinganine and sphingosine as well as the expression of enzymes related to de novo ceramide synthesis pathway. Moreover, in our study, we observed augmented ceramide and sphingomyelin efflux into the incubation media in these groups. In addition, RSV substantially reduced intracellular triacylglycerols accumulation in overload conditions.The above-mentioned findings suggest that RSV, at least partially, demonstrates a potential protective effect on HepG2 cells in a overload state.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: fat metabolism

Obesity-induced activation of JunD promotes myocardial accumulation and metabolic cardiomyopathy.

Metabolic cardiomyopathy (MC)-characterized by intra-myocardial triglyceride (TG) accumulation and lipotoxic damage-is an emerging cause of heart failure in obese patients. Yet, its mechanisms remain poorly understood. The Activator Protein 1 (AP-1) member JunD was recently identified as a key modulator of hepatic in obese mice. The present study investigates the role of JunD in obesity-induced MC.JunD transcriptional activity was increased in hearts from diet-induced obese (DIO) mice and was associated with myocardial TG accumulation and left ventricular (LV) dysfunction. Obese mice lacking JunD were protected against MC. In DIO hearts, JunD directly binds PPARγ promoter thus enabling transcription of genes involved in TG synthesis, uptake, hydrolysis, and storage (i.e. Fas, Cd36, Lpl, Plin5). Cardiac-specific overexpression of JunD in lean mice led to PPARγ activation, cardiac steatosis, and dysfunction, thereby mimicking the MC phenotype. In DIO hearts as well as in neonatal rat ventricular myocytes exposed to , Ago2 immunoprecipitation, and luciferase assays revealed JunD as a direct target of miR-494-3p. Indeed, miR-494-3p was down-regulated in hearts from obese mice, while its overexpression prevented lipotoxic damage by suppressing JunD/PPARγ signalling. JunD and miR-494-3p were also dysregulated in myocardial specimens from obese patients as compared with non-obese controls, and correlated with myocardial TG content, expression of PPARγ-dependent genes, and echocardiographic indices of LV dysfunction.miR-494-3p/JunD is a novel molecular axis involved in obesity-related MC. These results pave the way for approaches to prevent or treat LV dysfunction in obese patients.Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.

Keyword: fat metabolism

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: fat metabolism

Chitosan Oligosaccharides Improve Glucolipid Disorder in Liver by Suppression of Obesity-Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid disorder using (PA)-induced HepG2 cells and high- diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid -related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid -related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid -related diseases.

Keyword: fat metabolism

Permethrin and ivermectin modulate in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic . The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Effects of dietary perilla seed oil supplementation on , meat quality, and fatty profiles in Yellow-feathered chickens.

This study evaluated the effect of the dietary replacement of 1% lard (CT) with 1% perilla oil (PO), 0.9% perilla oil + 0.1% anise oil (PA), or 0.9% perilla oil + 0.1% ginger oil (PG) on indices of , antioxidant capacity, meat quality, and fatty profiles from Yellow-feathered chickens at day 63. Compared with the CT chickens, those given perilla oil had decreased (P < 0.05) plasma levels including triglycerides (TG), total cholesterol (TCH), and low-density lipoprotein cholesterol (LDL-C). Hepatic TG, TCH levels, and fatty synthase activity were also decreased (P < 0.05) in chickens fed diets containing perilla oil. Abdominal percentage was significantly decreased in birds fed the PG compared to CT diets. Birds fed the PA or PG diets had increased (P < 0.05) hepatic total SOD, glutathione peroxidase, and glutathione-S-transferase than in chickens given PO alone. In addition, the content of reduced glutathione (GSH) in breast muscle was lower (P < 0.05) in birds fed PO compared with those given PG, and the reverse was true for content of malondialdehyde. Compared with the CT diet, the PO diet decreased breast muscle shear values and increased yellowness (b*) of breast muscle (P < 0.05). Birds fed the PA or PG diets had meat with better overall acceptability than those fed the CT diet. Chickens fed perilla oil diets exhibited higher contents of α-linolenic (C18:3n-3), DHA (22:6n-3), polyunsaturated fatty acids, and n-3 fatty acids, together with a lower content of myristic (C14:0), (C16:0), stearic (C18:0), total saturated fatty acids, and n-6/n-3 ratio compared to controls (P < 0.05). These findings indicate that perilla oil has the potential to decrease -related indices and improve fatty profiles of breast meat in chickens without adverse effect on antioxidant status or meat quality; this was even better when perilla oil was given together with anise oil or ginger oil.© 2019 Poultry Science Association Inc.

Keyword: fat metabolism

Replication of Marek\'s Disease Virus Is Dependent on Synthesis of Fatty and Prostaglandin E.

Marek\'s disease virus (MDV) causes deadly lymphoma and induces an imbalance of the in infected chickens. Here, we discovered that MDV activates the fatty synthesis (FAS) pathway in primary chicken embryo fibroblasts (CEFs). In addition, MDV-infected cells contained high levels of fatty acids and showed increased numbers of droplets (LDs). Chemical inhibitors of the FAS pathway (TOFA and C75) reduced MDV titers by approximately 30-fold. Addition of the downstream metabolites, including malonyl-coenzyme A and , completely restored the inhibitory effects of the FAS inhibitors. Furthermore, we could demonstrate that MDV infection activates the COX-2/prostaglandin E (PGE) pathway, as evident by increased levels of arachidonic , COX-2 expression, and PGE synthesis. Inhibition of the COX-2/PGE pathway by chemical inhibitors or knockdown of COX2 using short hairpin RNA reduced MDV titers, suggesting that COX-2 promotes virus replication. Exogenous PGE completely restored the inhibition of the COX-2/PGE pathway in MDV replication. Unexpectedly, exogenous PGE also partially rescued the inhibitory effects of FAS inhibitors on MDV replication, suggesting that there is a link between these two pathways in MDV infection. Taken together, our data demonstrate that the FAS and COX-2/PGE pathways play an important role in the replication of this deadly pathogen. Disturbances of the in chickens infected with MDV contribute to the pathogenesis of disease. However, the role of in MDV replication remained unknown. Here, we demonstrate that MDV infection activates FAS and induces LD formation. Moreover, our results demonstrate that MDV replication is highly dependent on the FAS pathway and the downstream metabolites. Finally, our results reveal that MDV also activates the COX-2/PGE pathway, which supports MDV replication by activating PGE/EP2 and PGE/EP4 signaling pathways.Copyright © 2019 Boodhoo et al.

Keyword: fat metabolism

miR-21-5p regulates mitochondrial respiration and content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate content, peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the -induced increase in cellular content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of peroxidation, was significantly increased with treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both content and peroxidation in H9C2 cells. This likely occurs by reducing cellular uptake and utilization, shifting cellular toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: fat metabolism

Novel hypolipidemic conjugates of fatty and bile with lysine for linkage.

Novel fatty -bile conjugates (1a-1k) were designed and synthesized by coupling of the fatty acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve liver-specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high- diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced liver damage.

Keyword: fat metabolism

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced Lipogenesis by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells.

4EBP1 is a chief downstream factor of mTORC1, and PPARγ is a key lipogenesis-related transcription factor. mTORC1 and PPARγ are associated with . However, it is unknown which effector protein connects mTORC1 and PPARγ. This study investigated the interaction between 4EBP1 with PPARγ as part of the underlying mechanism by which insulin-induced synthesis and secretion are regulated by mTORC1 in primary bovine mammary epithelial cells (pBMECs). Rapamycin, a specific inhibitor of mTORC1, downregulated 4EBP1 phosphorylation and the expression of PPARγ and the following lipogenic genes: lipin 1, DGAT1, ACC, and FAS. Rapamycin also decreased the levels of intracellular triacylglycerol (TAG); 10 types of fatty ; and the accumulation of TAG, (PA), and stearic (SA) in the cell culture medium. Inactivation of mTORC1 by shRaptor or shRheb attenuated the synthesis and secretion of TAG and PA. In contrast, activation of mTORC1 by Rheb overexpression promoted 4EBP1 phosphorylation and PPARγ expression and upregulated the mRNA and protein levels of lipin 1, DGAT1, ACC, and FAS, whereas the levels of intracellular and extracellular TAG, PA, and SA also rose. Further, 4EBP1 interacted directly with PPARγ. Inactivation of mTORC1 by shRaptor prevented the nuclear location of PPARγ. These results demonstrate that mTORC1 regulates synthesis and secretion by inducing the expression of lipin 1, DGAT1, ACC, and FAS, which is likely mediated by the 4EBP1/PPARγ axis. This finding constitutes a novel mechanism by which synthesis and secretion are regulated in pBMECs.

Keyword: fat metabolism

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial .A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

miR-125a-5p ameliorates hepatic glycolipid disorder in type 2 diabetes mellitus through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 diabetes mellitus (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid in T2DM, which can inhibit hepatic lipogenesis and gluconeogenesis and elevate glycogen synthesis by targeting STAT3.

Keyword: fat metabolism

Dual Specificity Phosphatase 12 Regulates Hepatic Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high- diet (HFD)-induced and high- high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic . DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high- conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: fat metabolism

Alterations to the microbiota-colon-brain axis in high--diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated- diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated- diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: fat metabolism

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: fat metabolism

Cyclophilin D participates in the inhibitory effect of high- diet on the expression of steroidogenic acute regulatory protein.

The high- diet (HFD)-induced obesity is responsible for the testosterone deficiency (TD). However, the mechanism remains unknown. Mitochondrial homeostasis is proved to be important for maintaining the function of steroidogenic acute regulatory protein (StAR), the first rate-limiting enzyme in testosterone synthesis. As the key regulator of mitochondrial membrane permeability, cyclophilin D (CypD) plays a crucial role in maintaining mitochondrial function. In this study, we sought to elucidate the role of CypD in the expression of StAR affected by HFD.To analyse the influence of CypD on StAR in vivo and in vitro, mouse models of HFD, CypD overexpression and CypD knockout (Ppif ) as well as Leydig cells treated with (PA) and CypD overexpression plasmids were examined with an array of metabolic, mitochondrial function and molecular assays.Compared with the normal diet mice, consistent with reduced testosterone in testes, the expressions of StAR in both mRNA and protein levels in HFD mice were down-regulated, while expressions of CypD were up-regulated. High- intake impaired mitochondrial function with the decrease in StAR in Leydig cells. Overexpression of CypD inhibited StAR expressions in vivo and in vitro. Compared with C57BL/6 mice with HFD, expressions of StAR were improved in Ppif mice with HFD.Mitochondrial CypD involved in the inhibitory effect of HFD on StAR expression in testes.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fat metabolism

Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether obesity impacts placental transport and remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and insulin-sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal obesity. Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to insulin resistance and low-grade inflammation in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by obesity.-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

Keyword: fat metabolism

Deciphering the intervention mechanism of Taohong Siwu Decoction following the abnormal uterine bleeding rats based on serum metabolic profiles.

Abnormal uterine bleeding (AUB), one of the most significant characters of incomplete abortion, is a widespread phenomenon in gynecological that put a woman into a terrible physiological and psychological state. Taohong Siwu Decoction (TSD) is a traditional Chinese medicine (TCM) prescriptions which have treated AUB in China for decades. Our previous study elucidated that TSD reduced the volume of uterine bleedings as well as repaired the endometrium. The present study aims to investigate the mechanisms of TSD on AUB based on serum metabolomics. In this study, serum metabolic profile data was collected using ultra high-performance liquid chromatography with ion trap/time-of-flight mass spectrometry and gas chromatography-mass spectrometry. 23 potential biomarkers (urea, serine, L-proline, L-glutamic , , l-acetylcarnitine, LysoPC(16:0), LysoPC(20:4), l-proline, linoleic , stearic , l-isoleucine, phenylalanine, l-tyrosine, Oleic , et al) were eventually identified using multivariate statistical analysis (PCA and OPLS-DA) with VIP\u2009>\u20091, P\u2009<\u20090.05. Correlation analysis, fold-change (FC), area under receiver characteristic (ROC), false discovery rate (FDR) were used for data confirmation to ensure the authenticity of the data. The related-metabolic pathway mainly included amino (Phenylalanine, tyrosine, and tryptophan ; Valine, leucine and isoleucine biosynthesis; Arginine and proline ; Glycine, serine and threonine ) and (linoleic , glycerophospholipid ). The results show that TSD has a favorable therapeutic effect on AUB by adjusting the metabolic disorders, which could provide dietary guidance for the clinic.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic fatty liver disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of emulsion and sucrose for 9 weeks. The effects of QWTX on , liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

APOE and the Association of Fatty Acids With the Risk of Stroke, Coronary Heart Disease, and Mortality.

Background and Purpose- The role of dietary on cardiovascular health and mortality remains under debate. Because the APOE is central to the transport and of lipids, we examined associations between plasma fatty acids and the risk of stroke, coronary heart disease, and mortality by APOE-ε4 genotype. Methods- We included 943 FHS (Framingham Heart Study) and 1406 3C (Three-City) Bordeaux Study participants. Plasma docosahexaenoic, linoleic, arachidonic, and fatty acids were measured at baseline by gas chromatography. All-cause stroke, ischemic stroke, coronary heart disease, and all-cause mortality events were identified prospectively using standardized protocols. Each cohort used Cox models to separately relate fatty levels to the risk of developing each event during ≤10 years of follow-up adjusting for potential confounders and stratifying by APOE genotype (ε4 carriers versus noncarriers). We then meta-analyzed summary statistics using random-effects models. Results- On average, participants had a mean age of 74 years, 61% were women, and 21% (n=483) were APOE-ε4 carriers. Meta-analysis results showed that, only among APOE-ε4 carriers, every SD unit increase in linoleic was associated with a reduced risk of all-cause stroke (hazard ratio [HR], 0.54 [95% CI, 0.38-0.78]), ischemic stroke (HR, 0.48 [95% CI, 0.33-0.71]), and all-cause mortality (HR, 0.70 [95% CI, 0.57-0.85]). In contrast, every SD unit increase in was related to an increased risk of all-cause stroke (HR, 1.58 [95% CI, 1.16-2.17]), ischemic stroke (HR, 1.76 [95% CI, 1.26-2.45]), and coronary heart disease (HR, 1.48 [95% CI, 1.09-2.01]), also in APOE-ε4 carriers only. Results for docosahexaenoic and arachidonic were heterogeneous between cohorts. Conclusions- These exploratory results suggest that APOE-ε4 carriers may be more susceptible to the beneficial or adverse impact of fatty acids on cardiovascular disease and mortality. In this subgroup, higher linoleic was protective for stroke and mortality, whereas was a risk factor for stroke and coronary heart disease. The mechanisms underlying these novel findings warrant further investigation.

Keyword: fat metabolism

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host energy through the regulation of body storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

[Psoralen and isopsoralen improve disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: fat metabolism

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by -related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic accumulation in rats with high- diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of - and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of -related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of , suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Physicochemical characteristics of blend from hydrogenated coconut oil and acyl migrated palm mid-fraction.

Palm mid-fraction (PMF), which has a high content of symmetric POP, was converted to asymmetric PPO (APMF) via acyl migration. After solvent fractionation, the liquid phase of acyl migrated PMF (APMF-L) was obtained and blended with hydrogenated coconut oil (HCO, 50:50, w/w) to produce a blend (namely, an alternative blend) which had reduced saturated fatty content while having similar melting behavior to HCO. In an alternative blend, the major fatty acids were lauric (27.94), (26.93) and oleic (15.75\u202fmol%) . The solid index was quite similar to that of HCO, especially at 28-44\u202f°C. Nevertheless, an alternative blend had lower saturated fatty content, by 18%, compared to HCO. The content of highly atherogenic myristic was reduced by approximately 40%. The alternative blend in this study could be used as a raw material for non-dairy cream with low saturated content.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Analysis of Titin in Red and White Muscles: Crucial Role on Muscle Contractions Using a Fish Model.

Several studies have compared molecular components between red and white skeletal muscles in mammals. However, mammalian skeletal muscles are composed of mixed types of muscle fibers. In the current study, we analyzed and compared the distributions of titin, , phosphate ions, and fatty levels in red and white muscles using a fish model (), which is rich in red and white muscles, and these are well separated. Oil-red O staining showed that red muscle had more-abundant lipids than did white muscle. A time-of-flight secondary-ion mass spectrometric (TOF-SIMS) analysis revealed that red muscle possessed high levels of and oleic , but white muscle contained more phosphate ions. Moreover, elastica-van Gieson (EVG) and Mito-Tracker green FM staining showed that collagen and elastic fibers were highly, respectively, distributed in connective tissues and mitochondria in red muscle. An electron micrographic analysis indicated that red muscle had a relatively higher number of mitochondria and longer sarcomere lengths and Z-line widths, while myofibril diameters were thicker in white muscle. Myofibrillar proteins separated by SDS-PAGE showed that the major giant protein, titin, was highly expressed in white muscle than in red muscle. Furthermore, ratios of titin to myosin heavy chain (MHC) (titin/MHC) were about 1.3 times higher in white muscle than red muscle. We postulated that white muscle is fit for short and strong contractile performance due to high levels of titin and condensed sarcomeres, whereas red muscle is fit for low intensity and long-lasting activity due to high levels of lipids and mitochondria and long sarcomeres.

Keyword: fat metabolism

Thioacetamide potentiates high cholesterol and high diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH.

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and liver anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

A high- diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high- diet (HFD) commonly leads to obesity and insulin resistance. However, even 3\u2009days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus.Mice were fed either a low- diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: fat metabolism

Prepartum fatty supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb liver and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb liver and adipose tissue fatty (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and liver and adipose tissue collected for FA and gene expression analysis. Liver concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in liver tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), fatty synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in liver of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain fatty 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that lipogenesis decreased, and lipolysis increased in lamb liver with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that lipogenesis decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not liver, to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: fat metabolism

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased , protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and .

Keyword: fat metabolism

Adverse Effects in Skeletal Muscle Following the Medicinal Use of Nicotiana glauca.

Nicotiana glauca is a cosmopolitan shrub, used in medicine to treat swellings, wounds, sores and cancer. However, its users lack of knowledge of the adverse effects. We seek to evaluate the effects of extracts from N. glauca on myoblasts, identifying the compounds which cause undesirable effects. Myoblasts are important in muscle homeostasis, thus a high death rate of them cause myopathies. We performed an ethanolic extraction from leaves of N. glauca and the extract was successively partitioned with hexane, chloroform and ethyl acetate. The effects of extracts in C2C12 cells were analysed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Mitotracker and 4\',6-diamidino-2-phenylindole (DAPI) staining, Western blotting, real-time PCR and immunofluorescence assays. Caspase activity was studied. The fraction with the highest apoptotic effects was analysed by chromatography, NMR and GC-MS spectrometry were used to identify the apoptotic agent, after which its biological activity was evaluated. The extracts from N. glauca induced apoptosis in C2C12 cells involving caspase-3/7. We found that the extracts trigger a defence response in muscle through Akt and heat shock protein 27 (HSP27). We identified an apoptotic agent as . These data suggest that the use of N. glauca in hormone replacement therapy, or in other therapies affects skeletal muscle homeostasis, worsening the negative effects of the menopause. Thus, the relevance of this work lies in the fact that it is the first time that a report about the molecular mechanism responsible for the side effects of medicinal use of N. glauca, has been shown. Moreover the compound responsible for these effects has been identified.

Keyword: fat metabolism

Human cytochrome P450 enzymes bind drugs and other substrates mainly through conformational-selection modes.

Cytochrome P450 (P450) enzymes are major catalysts involved in the oxidations of most drugs, steroids, carcinogens, -soluble vitamins, and natural products. The binding of substrates to some of the 57 human P450s and other mammalian P450s is more complex than a two-state system and has been proposed to involve mechanisms such as multiple ligand occupancy, induced-fit, and conformational-selection. Here, we used kinetic analysis of binding with multiple concentrations of substrates and computational modeling of these data to discern possible binding modes of several human P450s. We observed that P450 2D6 binds its ligand rolapitant in a mechanism involving conformational-selection. P450 4A11 bound the substrate lauric via conformational-selection, as did P450 2C8 with . Binding of the steroid progesterone to P450 21A2 was also best described by a conformational-selection model. Hexyl isonicotinate binding to P450 2E1 could be described by either a conformational-selection or an induced-fit model. Simulation of the binding of the ligands midazolam, bromocriptine, testosterone, and ketoconazole to P450 3A4 was consistent with an induced-fit or a conformational-selection model, but the concentration dependence of binding rates for varying both P450 3A4 and midazolam concentrations revealed discordance in the parameters, indicative of conformational-selection. Binding of the P450s 2C8, 2D6, 3A4, 4A11, and 21A2 was best described by conformational-selection, and P450 2E1 appeared to fit either mode. These findings highlight the complexity of human P450-substrate interactions and that conformational-selection is a dominant feature of many of these interactions.© 2019 Guengerich et al.

Keyword: fat metabolism

Mechanisms of electrical remodeling in lipotoxic guinea pig heart.

To develop an adult guinea pig model of lipotoxicity and explore the underlying mechanisms associated with changes in the expression of the delayed rectifier potassium current (I).Lipotoxicity may represent a common link among metabolic disorders and a higher vulnerability to arrhythmias.Whole-cell patch clamp, and (PA, a potent inducer of lipotoxicity), were used to assess mechanisms of short-term (∼50 days) high- diet (HFD) feeding on atrial electrophysiology in guinea pig hearts and myocytes.HFD fed guinea pigs were significantly heavier, displayed hypertriglyceridemia and hypercholesterolemia; but no signs of hyperglycemia or inflammation compared to low- diet fed controls. Increasing cardiac PA levels, resulted in shortened atrial action potential duration, and increased I density. Inhibition of phosphoinositide 3-kinase (PI3K) prevented increases in I due to PA. Acute (≥1hr) exposure of atrial myocytes to exogenous PA (1\u202fmM) increased the density of the rapid delayed rectifier potassium current I, while it was decreased with the unsaturated oleic (OA, 1\u202fmM). Serine-threonine protein phosphatase-2 (PP2A) inhibition with cantharidin reversed the effect of OA on I.Our data provide evidence of a novel lipotoxic guinea pig model with signs of vulnerability to arrhythmias. Inhibition of PA/PI3K/I and/or activation of the OA/PP2A/I pathways may be therapeutically beneficial for lipotoxic arrhythmias.Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The inhibition of Nrf2 accelerates renal deposition through suppressing the ACSL1 expression in obesity-related nephropathy.

Obesity has become a worldwide epidemic, and the incidence of obesity is increasing year by year. Obesity-related nephropathy (ORN) is a common kidney complication of obesity. Long-chain acyl-CoA synthetases-1, (ACSL1), is a key enzyme in the oxidative of fatty acids in mitochondria and ACSL1 may play a direct role in renal deposition and promote the progress of ORN. In this study, we focus on the renoprotective role of ACSL1 in ORN. Electron microscopy, immunohistochemical (IHC) staining, Western blot, and real-time PCR were used to detect the expression of ACSL1and Nrf2 in ORN patients, ob/ob mice and (PA)-treated HK-2 cells. Oil red staining and Elisa Kit were used to detect the intracellular FFA and TG contents in ob/ob mice and PA-treated HK-2 cells. Dihydroethidium (DHE) staining and the MDA/SOD measurement were used to detect the ROS production. In order to demonstrate the role of ACSL1 and the interaction between ACSL1 and Nrf2 in ORN, related siRNA and plasmid were transfected into HK-2 cells. More ROS production and renal deposition have been found in ORN patients, ob/ob mice and PA-treated HK-2 cells. Compared with control, all the expression of ACSL1and Nrf2 were down-regulated in ORN patients, ob/ob mice and PA-treated HK-2 cells. The Nrf2 could regulate the expression of ACSL1 and the ACSL1 played the direct role in renal deposition. The Nrf2 is inhibited in ORN, resulting more ROS production and oxidative stress. Increased oxidative stress will suppress the expression of ACSL1, which could increase the intracellular FFA and TG contents, ultimately leading to renal deposition in renal tubulars and accelerating the development of ORN.

Keyword: fat metabolism

Inhibition of soluble epoxide hydrolase attenuates a high- diet-mediated renal injury by activating PAX2 and AMPK.

A high- diet (HFD) causes obesity-associated morbidities involved in macroautophagy and chaperone-mediated autophagy (CMA). AMPK, the mediator of macroautophage, has been reported to be inactivated in HFD-caused renal injury. However, PAX2, the mediator for CMA, has not been reported in HFD-caused renal injury. Here we report that HFD-caused renal injury involved the inactivation of Pax2 and Ampk, and the activation of soluble epoxide hydrolase (sEH), in a murine model. Specifically, mice fed on an HFD for 2, 4, and 8 wk showed time-dependent renal injury, the significant decrease in renal Pax2 and Ampk at both mRNA and protein levels, and a significant increase in renal sEH at mRNA, protein, and molecular levels. Also, administration of an sEH inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea, significantly attenuated the HFD-caused renal injury, decreased renal sEH consistently at mRNA and protein levels, modified the renal levels of sEH-mediated epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) as expected, and increased renal Pax2 and Ampk at mRNA and/or protein levels. Furthermore, (PA) treatment caused significant increase in , and decrease in both Pax2 and Ampk in murine renal mesangial cells (mRMCs) time- and dose-dependently. Also, 14(15)-EET (a major substrate of sEH), but not its sEH-mediated metabolite 14,15-DHET, significantly reversed PA-induced increase in , and PA-induced decrease in Pax2 and Ampk. In addition, plasmid construction revealed that Pax2 may positively regulate Ampk transcriptionally in mRMCs. This study provides insights into and therapeutic target for the HFD-mediated renal injury.

Keyword: fat metabolism

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: fat metabolism

Outdoor cultivation of the green microalga Chlorella vulgaris under stress conditions as a feedstock for biofuel.

The present work investigated the potential of the green alga Chlorella vulgaris to produce high-quality biofuel under culture stress conditions. The cultivation was carried out in a 1000 l\xa0open plate tank system, which provides biomass yields comparable to open pond systems, but with less area needed. Algal biomass and content were measured repeatedly. We compared the two solvent systems n-hexane and hexane/isopropanol (HIP) for extraction efficiency of lipids and applied three different extraction methods Soxhlet, soaking, and soaking followed by Soxhlet (soak-Sox). The combination of the HIP solvent and the soak-Sox provided the highest yield (15.8\u2009±\u20090.174). Volumetric biomass and productivity were 0.201\xa0g\xa0l\xa0day and 31.71\xa0mg\xa0l\xa0day, respectively, whereas areal biomass and productivity were 25.73\xa0g\xa0m\xa0day and 4.066\xa0g\xa0m\xa0day, respectively. The fatty profile by means of gas chromatography resulted in seven fatty acids from C to C. The most abundant fatty methyl esters (FAMES) were (C16:0), oleic (C18:1), and stearic (C18:0) acids. synthesis enhanced by optimizing the Kuhl growth medium with replacing nitrate by urea (50% N compared to the original recipe) increased salt content (10\xa0g/l NaCl), ferrous sulfate (0.5\xa0g/l), and sodium acetate addition (1\xa0g/l). With regard to density, kinematic viscosity, gravity, pour point, flash point, and cetane number, the Chlorella-biodiesel comply with ASTM and EN standards thus pointing at the high potential of lipids synthesized by Chlorella as a feedstock for biodiesel production.

Keyword: fat metabolism

[Exploration about the protection mechanism of 5-hydroxy-1-methylhydantoin on paraquat poisoning model].

To investigate the effects of 5-hydroxy-1-methylhydantoin (HMH) on kidney injury induced by paraquat (PQ).Fifteen SPF healthy Kunming mice were randomly divided into normal saline (NS) control group, PQ poisoning model group and HMH intervention group, with 5 mice in each group. PQ poisoning model was challenged by one-time gavage of 30 mg/kg PQ solution. The NS group received the same amount of NS by gavage. The HMH group was given 100 mg/kg of HMH immediately after the model was made and continued to be gavaged. Mice in each group were sacrificed 1 day after HMH gavage and heart blood and renal tissue were harvested for examination. The morphological changes of renal tissue were observed under light microscope by hematoxylin-eosin (HE) staining. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in renal tissue were detected according to the instructions of the kit. The expression of heme oxygenase-1 (HO-1) and interleukin-1β (IL-1β) in renal tissues were detected by Western Blot. The serum metabolites were detected by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), the overall distribution of each sample was observed by principal component analysis (PCA), the accuracy of the model was evaluated by multidimensional analysis orthogonal partial least squares-discriminant analysis (OPLS-DA), and the difference metabolites were screened by variable importance in the projection (VIP) value > 1.Light microscopic observation showed that: glomerular structure in NS group was clear, there was no hyperemia and inflammatory cell infiltration in renal interstitium and blood vessels. In PQ group, some glomeruli atrophy and necrosis, capillary congestion in glomeruli, infiltration of inflammatory cells around glomeruli, swelling of renal tubular epithelial cells, slight stenosis of lumen, and occasional necrosis and exfoliation of epithelial cells occurred. The degree of kidney injury in HMH group was significantly less than that in PQ group. Compared with the NS group, the content of MDA in the PQ group was significantly increased (nmol/g: 6.70±0.84 vs. 2.70±0.43, P < 0.01) and the activity of SOD was significantly decreased (kU/L: 33.30±4.66 vs. 50.20±3.23, P < 0.05), the protein expression of HO-1 and IL-1β were significantly increased (HO-1/β-actin: 1.11±0.12 vs. 0.61±0.13, IL-1β/β-actin: 0.93±0.13 vs. 0.32±0.06, both P < 0.05). Compared with the PQ group, the content of MDA in the HMH group was significantly decreased (nmol/g: 5.10±0.93 vs. 6.70±0.84, P < 0.05) and the activity of SOD was significantly increased (kU/L: 61.00±9.02 vs. 33.30±4.66, P < 0.05), the protein expression of HO-1 was significantly decreased (HO-1/β-actin: 0.77±0.07 vs. 1.11±0.12, P < 0.05), however, there was no significant difference in the protein expression of IL-1β (IL-1β/β-actin: 0.87±0.13 vs. 0.93±0.13, P > 0.05). Metabolite detection results showed that: compared with NS group, the levels of creatinine, glycine, succinic , fumaric and citric were significantly increased in the PQ group (VIP value was 1.50, 1.58, 1.64, 1.74 and 1.95 respectively, all P < 0.05), while the levels of ,α-tocopherol and 6-phosphogluconic were significantly decreased (VIP value was 1.10, 1.55 and 1.56 respectively, all P < 0.05). Compared with the PQ group, the levels of creatinine and citric were significantly decreased in the HMH group (VIP value was 1.50 and 1.86, both P < 0.05), while trans-4-hydroxy-proline, D-glyceric , 2, 6-fructose phosphate, 6-phosphate gluconic and aminomalonic were significantly increased (VIP value was 1.36, 1.55, 1.63, 1.68 and 1.76 respectively, all P < 0.05).HMH protects kidney injury caused by PQ poisoning by correcting tricarboxylic acids cycle disturbance, peroxidation and energy disturbance, and its mechanism is related to the regulation of HO-1 protein expression through Nrf2 pathway.

Keyword: fat metabolism

Berberine attenuates nonalcoholic hepatic steatosis through the AMPK-SREBP-1c-SCD1 pathway.

Berberine (BBR), a natural compound extracted from Chinese herb, has been shown to effectively attenuate nonalcoholic fatty liver disease (NAFLD) in clinic. However, the mechanism underlying the effect of BBR is not fully understood. Stearyl-coenzyme A desaturase 1 (SCD1) mediates in liver. Therefore, we hypothesized that SCD1 mediated the beneficial effect of BBR on NAFLD.The expression of SCD1 was measured in the liver of NAFLD patients and ob/ob mice. The effect of BBR on NAFLD was evaluated in C57BL/6\u202fJ mice on high diet (HFD). The effect of BBR was also investigated in HepG2 and AML12\u202fcells exposed to high glucose and . Oil red O staining was performed to detect triglyceride (TG) level. Quantitative real-time polymerase chain reaction and Western blot were used to detect the messenger ribonucleic (mRNA) and protein expression of target genes. The activity of SCD1 promoter was measured by dual-luciferase reporter assay.The expression of SCD1 was increased in the liver of NAFLD patients and ob/ob mice. BBR reduced hepatic TG accumulation and decreased the expressions of hepatic SCD1 and other TG synthesis related genes both in vivo and in vitro. Knockdown of SCD1 expression mimicked the effect of BBR decreasing TG level in steatotic hepatocytes, whereas overexpression of SCD1 attenuated the effect of BBR. Mechanistically, BBR promoted the phosphorylation of AMP-activated protein kinase (AMPK) and sterol regulatory element-binding protein-1c (SREBP-1c) in HepG2 cells and the liver of HFD-fed mice. Activation of the AMPK-SREBP-1c pathway and sterol regulatory element (SRE) motif in SCD1 promoter (-920/-550) was responsible for the BBR-induced suppression of SCD1.BBR reduces liver TG synthesis and attenuates hepatic steatosis through the activation of AMPK-SREBP-1c-SCD1 pathway.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high energy value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on are still poorly understood. Therefore, this study evaluated the fatty (FA) profile and the effects of baru oil on liver and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and liver. The results demonstrated that animals developed marked hypercholesterolemia, liver steatosis, and increased peroxidation in the aorta. Treatment with baru oil attenuated peroxidation and drastically reduced liver damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: fat metabolism

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related metabolic diseases such as type II diabetes mellitus (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: fat metabolism

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/ in type 2 diabetes mellitus. Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high- diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

Keyword: fat metabolism

Gut Microbiota and Metabolome Response of Seed Oil on Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut microbiota dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan .

Keyword: fat metabolism

mTORC2 Regulates Lipogenic Gene Expression through PPAR to Control Synthesis in Bovine Mammary Epithelial Cells.

The mechanistic target of rapamycin complex 2 (mTORC2) primarily functions as an effector of insulin/PI3K signaling to regulate cell proliferation and is associated with cell . However, the function of mTORC2 in is not well understood. In the present study, mTORC2 was inactivated by the ATP-competitive mTOR inhibitor AZD8055 or shRNA targeting in primary bovine mammary epithelial cells (pBMECs). MTT assay was performed to examine the effect of AZD8055 on cell proliferation. ELISA assay and GC-MS analysis were used to determine the content of . The mRNA and protein expression levels were investigated by RT/real-time PCR and western blot analysis, respectively. We found that cell proliferation, mTORC2 activation, and secretion were inhibited by AZD8055. was knocked down and mTORC2 activation was specifically attenuated by the shRNA. Compared to control cells, the expression of the transcription factor gene and the lipogenic genes , , , and was downregulated in silencing cells. As a result, the content of intracellular triacylglycerol (TAG), (PA), docosahexaenoic (DHA), and other 16 types of fatty was decreased in the treated cells; the accumulation of TAG, PA, and DHA in cell culture medium was also reduced. Overall, mTORC2 plays a critical role in regulating lipogenic gene expression, synthesis, and secretion in pBMECs, and this process probably is through PPAR. This finding provides a model by which lipogenesis is regulated in pBMECs.

Keyword: fat metabolism

Dietary modulation of energy homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: fat metabolism

Influence of nutrient formulations on growth, yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.

The study was conducted to analyse the influence of three nutrient formulations, namely BG-11 medium, BBM and TAP medium, on growth potential and yield of two microalgal genera (Botryococcus sp. and Chlorella sp.) and to study the roles of N, P and other major nutrients. The study focussed on the general patterns of starch and synthesis and storage and to further assess how photosynthetic carbon partitioning into starch and is altered by conditions in growth media such as N and C presence as seen in BG11 medium which are known to induce neutral production and the lack of it in BBM and TAP medium. BG-11 medium performed better as compared to BBM and TAP medium in terms of biomass productivity and yield. The yield was highest in Botryococcus sp. (63.03% dry wt.) and Chlorella sp. (50.27% dry wt.) at 30th day of incubation. Mean biomass productivity was highest for Botryococcus in BBM medium (6.14\xa0mg/L/day) and for Chlorella in BG-11 medium (4.97\xa0mg/L/day). Mean productivity (50.78% and 39.36%) was highest in BG11 medium for both Botryococcus and Chlorella species, respectively. A sharp decline in sugar content was observed in the late stationary phase of growth from 30th day to 45th day. Fatty methyl ester (FAME) profile of the extracted lipids showed predominantly oleic , followed by and stearic in both the strains when grown in BG-11 medium. The other biodiesel quality parameters were in accordance with the international standards. A complex relationship was found between chemical composition and biodiesel properties. Proximity analysis indicated that the fuel properties of biodiesels are determined by a number of parameters and by the combination of different chemical compositions. The results provide an insight into organic carbon partitioning into compounds and how the organism\'s changes due to N-deplete culturing in TAP medium and inorganic carbon source availability as seen in BG-11 and BBM medium.

Keyword: fat metabolism

Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration.

Obesity may promote intervertebral disc degeneration (IDD) by non-mechanical means, by influencing levels of free fatty acids which could impair cell . This study aims to establish metabolic factors in obesity-related IDD independent of mechanical loading. In clinical study, we retrospectively reviewed 128 volunteers (73 males, 55 females, aged 29-88 years) and compared their grades of disk degeneration with obesity-related factors such as body weight, BMI, and serum levels. Clinically, the IDD group showed increased age, BMI and serum triglyceride. Triglyceride was a significant risk factor for IDD even after correction for BMI and age (P = 0.007). In obesity animal model, rats were fed a high- diet (HFD) in order to study its effects on disk and apoptosis. HFD rats had significantly higher serum levels of lipids, including triglyceride and non-esterified fatty , and showed significantly decreased markers of anabolism, increased catabolism and apoptosis in disk. Finally, rat nucleus pulposus (NP) cells were stimulated with a fatty (, PA) to gauge its effects on cell and apoptosis. Cell culture studies showed that NP cells exposed to PA showed increased apoptosis for activation of caspase 3, 7, 9, and PARP, which was primarily via the MAPK signal pathway, especially ERK pathway. In conclusion, hypertriglyceridemia can lead to IDD, independently of age and BMI. Hypertriglyceridemia appears to mediate disk cell apoptosis and matrix catabolism primarily via the ERK pathway.Copyright © 2019 Zhang, Chen, Huang, Wang, Shan, Liu, Chen, Li, Fan and Zhao.

Keyword: fat metabolism

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in . We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high- diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fat metabolism

Amelioration of Endoplasmic Reticulum Stress by Mesenchymal Stem Cells via Hepatocyte Growth Factor/c-Met Signaling in Obesity-Associated Kidney Injury.

Recent advances in the understanding of suggest a critical role of endoplasmic reticulum (ER) stress in obesity-induced kidney injury. Hepatocyte growth factor (HGF) is a pleiotropic cytokine frequently featured in stem cell therapy with distinct renotropic benefits. This study aims to define the potential link between human induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs)/bone marrow-derived MSCs (BM-MSCs) and ER stress in lipotoxic kidney injury induced by (PA) in renal tubular cells and by high- diet (HFD) in mice. iPS-MSCs or BM-MSCs alleviated ER stress (by preventing induction of Bip, chop, and unfolded protein response), inflammation (Il6, Cxcl1, and Cxcl2), and apoptosis (Bax/Bcl2 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells) in renal cortex of animals exposed to HFD thus mitigating histologic damage and albuminuria, via activating HGF/c-Met paracrine signaling that resulted in enhanced HGF secretion in the glomerular compartment and c-Met expression in the tubules. Coculture experiments identified glomerular endothelial cells (GECs) to be the exclusive source of glomerular HGF when incubated with either iPS-MSCs or BM-MSCs in the presence of PA. Furthermore, both GEC-derived HGF and exogenous recombinant HGF attenuated PA-induced ER stress in cultured tubular cells, and this effect was abrogated by a neutralizing anti-HGF antibody. Taken together, this study is the first to demonstrate that MSCs ameliorate lipotoxic kidney injury via a novel microenvironment-dependent paracrine HGF/c-Met signaling mechanism to suppress ER stress and its downstream pro-inflammatory and pro-apoptotic consequences. Stem Cells Translational Medicine 2019;8:898&910.© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

Keyword: fat metabolism

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high--diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: fat metabolism

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic insulin resistance and Type 2 diabetes (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of insulin resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo lipogenesis and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve adipogenesis through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with insulin resistance and risk of developing T2D.

Keyword: fat metabolism

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis (NASH) is characterized by accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on NASH and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce NASH and liver fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in inflammation and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated accumulation, inflammation and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents NASH and fibrosis via inhibition of oxidative stress and inflammation, highlighting polydatin as a potential therapeutic agent for prevention and treatment of NASH.

Keyword: fat metabolism

Free fatty -induced histone acetyltransferase activity accelerates accumulation in HepG2 cells.

Non-alcoholic fatty liver disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and in the liver have not been thoroughly investigated.Following oleic- (OPA)-induced accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: fat metabolism

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines , and gut microbiome-derived . Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: fat metabolism

Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Droplets in Cultured Hepatocytes.

A fluorescence microscopic method for characterizing size, quantity, and oxidation of droplets (LDs) in HepG2 cells was developed. LDs were induced by (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and peroxides. Each fatty increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2\'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.

Keyword: fat metabolism

of 13C-Labeled Fatty Acids in Term Human Placental Explants by Liquid Chromatography-Mass Spectrometry.

Placental transport and are poorly understood despite the importance for fetal development and lifelong health. We aimed to explore fatty (FA) processing in human villous placental explants from seven uncomplicated term singleton pregnancies delivered by elective cesarean section. Explants were treated with stable isotope-labeled (13C-PA), oleic (13C-OA), or docosahexaenoic (13C-DHA) for 3, 24, or 48 hours. Stable isotope-labeled lipids synthesized by placental explants from labeled FA were quantified, alongside endogenous unlabeled placental lipids, by liquid chromatography-mass spectrometry. Labeled phosphatidylcholines (PCs), triacylglycerols (TAGs), and phosphatidylethanolamines were detected in explants, whereas labeled lysophosphatidylcholines were found in both explants and conditioned media. 13C-PA was primarily directed into PC synthesis (74% of 13C-PA-labeled lipids), whereas 13C-OA was directed almost equally into PC and TAG synthesis (45% and 53%, respectively, of 13C-OA-labeled lipids). 13C-DHA was only detectable in TAGs. TAGs demonstrated the highest isotopic enrichment for all 13C-FAs with 13C-OA-TAGs comprising >50% of total OA-TAGs (unlabeled and labeled), consistent with TAGs being a labile and accessible reservoir for FA storage. Variations in incorporation were correlated to maternal glycemia and body mass index, suggesting that this experimental model could be used to investigate the effect of maternal factors on placental . We conclude that metabolic partitioning of freshly imported FAs into labile and less labile reservoirs in placenta is FA dependent. This process may partly mediate the physiological preferential transplacental transfer of particular FAs to the fetus, but may also be implicated in the fetoplacental pathophysiology of maternal metabolic dysfunction.Copyright © 2019 Endocrine Society.

Keyword: fat metabolism

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti-obesity remedy. Several pharmacological studies have shown its beneficial effects in . Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate and glucose and are therapeutic targets for type 2 diabetes. This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fat metabolism

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid and decrease hepatic insulin resistance development in a overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Bioconversion of barley straw lignin into biodiesel using Rhodococcus sp. YHY01.

Rhodococcus sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize p-coumaric , cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38\u202fg dcw/L, 0.25\u202fg dcw/L and 0.1\u202fg dcw/L, and accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC) value for p-coumaric (13.4\u202fmM), cresol (7.9\u202fmM), and 2,6 dimethoxyphenol (3.4\u202fmM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for Rhodococcus sp. YHY01 and resulted in 0.130\u202fg dcw/L with 39% w/w accumulation. Major fatty acids were (C16:0) 51.87%, palmitoleic (C16:l) 14.90%, and oleic (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Silibinin ameliorates hepatic accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: fat metabolism

Furanoid F- F6 Uniquely Induces NETosis Compared to C16 and C18 Fatty Acids in Human Neutrophils.

Various biomolecules induce neutrophil extracellular trap (NET) formation or NETosis. However, the effect of fatty acids on NETosis has not been clearly established. In this study, we focused on the NETosis-inducing ability of several molecules. We extracted the molecules present in Arabian Gulf catfish (, Val) skin gel, which has multiple therapeutic activities. Gas chromatography⁻mass spectrometry (GC-MS) analysis of the fraction-3 from the gel with NETosis-inducing activity contained fatty acids including a furanoid F- (F6; 12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic ) and common long-chain fatty acids such as (PA; C16:0), palmitoleic (PO; C16:1), stearic (SA; C18:0), and oleic (OA; C18:1). Using pure molecules, we show that all of these fatty acids induce NETosis to different degrees in a dose-dependent fashion. Notably, F6 induces a unique form of NETosis that is rapid and induces reactive oxygen species (ROS) production by both NADPH oxidase (NOX) and mitochondria. F6 also induces citrullination of histone. By contrast, the common fatty acids (PA, PO, SA, and OA) only induce NOX-dependent NETosis. The activation of the kinases such as ERK (extracellular signal-regulated kinase) and JNK (c-Jun N-terminal kinase) is important for long-chain fatty -induced NETosis, whereas, in F--induced NETosis, Akt is additionally needed. Nevertheless, NETosis induced by all of these compounds requires the final chromatin decondensation step of transcriptional firing. These findings are useful for understanding F-- and other fatty -induced NETosis and to establish the active ingredients with therapeutic potential for regulating diseases involving NET formation.

Keyword: fat metabolism

Genomics of -laden human hepatocyte cultures enables drug target screening for the treatment of non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a major health burden in need for new medication. To identify potential drug targets a genomic study was performed in -laden primary human hepatocyte (PHH) and human hepatoma cell cultures.PHH, HuH7 and HepG2 hepatoma cell cultures were treated with lipids and/or TNFα. Intracellular load was quantified with the ORO assay. The Affymetrix HG-U133+ array system was employed to perform transcriptome analysis. The droplet (LD) growth and fusion was determined by fluorescence microscopy. LD associated proteins were imaged by confocal immunofluorescence microscopy and confirmed by Western immunoblotting. Bioinformatics defined perturbed metabolic pathways.Whole genome expression profiling identified 227, 1031 and 571 significant regulated genes. Likewise, the combined and TNFα treatment of PHH, HuH7 and HepG2 cell cultures revealed 154, 1238 and 278 differentially expressed genes. Although genomic responses differed among in-vitro systems, commonalities were ascertained by filtering the data for LD associated gene regulations. Among others the LD-growth and fusion associated cell death inducing DFFA like effector C (CIDEC), perilipins (PLIN2, PLIN3), the synaptosome-associated-protein 23 and the vesicle associated membrane protein 3 were strongly up-regulated. Likewise, the PPAR targets pyruvate-dehydrogenase-kinase-4 and angiopoietin-like-4 were up-regulated as was hypoxia-inducible droplet-associated (HILPDA), flotilin and FGF21. Their inhibition ameliorates triglyceride and cholesterol accumulation. TNFα treatment elicited strong induction of the chemokine CXCL8, the kinases MAP3K8, MAP4K4 and negative regulators of cytokine signaling, i.e. SOCS2&SOCS3. Live cell imaging of DsRED calreticulin plasmid transfected HuH7 cells permitted an assessment of LD growth and fusion and confocal immunofluorescence microscopy evidenced induced LD-associated PLIN2, CIDEC, HIF1α, HILPDA, JAK1, PDK4 and ROCK2 expression. Notwithstanding, CPT1A protein was repressed to protect mitochondria from overload. Pharmacological inhibition of the GTPase-dynamin and the fatty transporter-2 reduced uptake by 28.5 and 35%, respectively. Finally, a comparisons of in-vitro/NAFLD patient biopsy findings confirmed common gene regulations thus demonstrating clinical relevance.The genomics of -laden hepatocytes revealed LD-associated gene regulations and perturbed metabolic pathways. Immunofluorescence microscopy confirmed expression of coded proteins to provide a rationale for therapeutic intervention strategies. Collectively, the in-vitro system permits testing of drug candidates.

Keyword: fat metabolism

Transcriptome analysis of elaiosomes and seeds provide insights into fatty biosynthesis.

Elaiosomes are specialized fleshy and edible seed appendages dispersed by ants. Lipids are the primary components of elaiosomes. is a well-known plant, the seeds of which are dispersed by ants. Previous studies have identified the presence of primary fatty acids in its elaiosomes and seeds. However, the molecular mechanisms underlying fatty biosynthesis in elaiosomes remain unknown.In order to gain a comprehensive transcriptional profile of the elaiosomes and seeds of , and understand the expression patterns of genes associated with fatty biosynthesis, four different developmental stages, including the flower-bud (Ch01), flowering (Ch02), young seed (Ch03), and mature seed (Ch04) stages, were chosen to perform whole-transcriptome profiling through the RNA-seq technology (Illumina NGS sequencing).A total of 63,064 unigenes were generated from 12 libraries. Of these, 7,323, 258, and 11,540 unigenes were annotated with 25 Cluster of Orthologous Groups, 43 Gene Ontology terms, and 373 Kyoto Encyclopedia of Genes and Genomes pathways, respectively. In addition, 322 genes were involved in transport and , and 508 genes were involved in the pathways. A total of 41 significantly differentially expressed genes (DEGs) involved in the pathways were identified, most of which were upregulated in Ch03 compared to Ch02, indicating that fatty biosynthesis primarily occurs during the flowering to the young seed stages. Of the DEGs, acyl-ACP thioesterases, acyl carrier protein desaturase (), and malonyl CoA-ACP transacylase were involved in synthesis; stearoyl-CoA desaturase and were involved in oleic synthesis, and acyl- omega-6 desaturase was involved in linoleic synthesis.

Keyword: fat metabolism

Downregulates Thyroglobulin (Tg), Sodium Iodide Symporter (NIS), and Thyroperoxidase (TPO) in Human Primary Thyrocytes: A Potential Mechanism by Which Lipotoxicity Affects Thyroid?

Our previous studies suggested that the thyroid might be a target organ affected by lipotoxicity, which might be partially related to the increasing prevalence of subclinical hypothyroidism. However, the underlying molecular mechanism is not yet clearly established. This study aimed to assess the effect of stimulation on thyrocyte function. Upon stimulation, intracellular contents of lipids, as well as the expression and activity of three key molecules in thyroid hormone synthesis (i.e., thyroglobulin, sodium iodide symporter, and thyroperoxidase), were determined in human primary thyrocytes. The contents of BODIPY® FL C16 (the fluorescently labeled analogue) entering into the thyrocytes were gradually increased with time extending. Accordingly, the intracellular accumulation of both triglyceride and free fatty acids increased in dose- and time-dependent manners. The effect of stimulation on thyroid hormone synthesis was then determined. Both the mRNA and protein levels of thyroglobulin, sodium iodide symporter, and thyroperoxidase were decreased following stimulation. Consistently, upon stimulation, the secreted thyroglobulin levels in supernatants, I uptake, and extracellular thyroperoxidase activity were all decreased in a dose-dependent manner. Our results demonstrated that upon stimulation, the expressions of the key molecules (thyroglobulin, sodium iodide symporter, and thyroperoxidase) were reduced and their activities were suppressed, which might lead to impaired thyroid hormone synthesis.

Keyword: fat metabolism

PAQR3 regulates phosphorylation of FoxO1 in insulin-resistant HepG2 cells via NF-κB signaling pathway.

Insulin resistance is a significant feature of type 2 diabetes mellitus and glucose and disorders. Activation of NF-κB signaling pathway plays an important role in the formation of insulin resistance. FoxO1 plays a major role in regulating glucose and , as well as insulin signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under insulin resistance. In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced insulin-resistant HepG2 cells, thereby causing glucose and disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced insulin-resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and disorders induced by insulin resistance.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial energy supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin . We further demonstrated the impact of cellular , and particularly cardiolipin , during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: fat metabolism

Uncommon properties of biosynthesis of isolated plastids in the unicellular red alga .

Red algae are a large group of photosynthetic eukaryotes that diverged from green algae over one billion years ago, and have various traits distinct from those of both green algae and land plants. Although most red algae are marine species (both unicellular and macrophytic), the Cyanidiales class of red algae includes unicellular species which live in hot springs, such as , which is a model species for biochemical and molecular biological studies. in red algae has previously been studied in intact cells. Here, we present the results of radiolabeling and stable isotope labeling experiments in intact plastids isolated from the unicellular red alga . We focused on two uncommon features: First, the galactose moiety of monogalactosyldiacylglycerol was efficiently labeled with bicarbonate, indicating that an unknown pathway for providing UDP-galactose exists within the plastid. Second, saturated fatty acids, namely, and stearic acids, were the sole products of fatty synthesis in the plastid, and they were efficiently exported. This finding suggests that the endoplasmic reticulum is the sole site of desaturation. We present a general principle of red algal biosynthesis, namely, \'indigenous C18 fatty acids are neither desaturated nor directly utilized within the plastid\'. We believe that this is valid in both lacking polyunsaturated fatty acids and marine red algae with a high content of arachidonic and eicosapentaenoic acids.

Keyword: fat metabolism

Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk content (3.60 vs. 3.41%), milk yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% -corrected milk (47.6 vs. 46.5 kg/d), and energy-corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk yield (1.70 vs. 1.66 kg/d), and 3.5% -corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk yield, energy-corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose in the obese mice.

Keyword: fat metabolism

Mp1 Protein, a Novel Virulence Factor, Carries Two Arachidonic -Binding Domains To Suppress Inflammatory Responses in Hosts.

infection causes talaromycosis (previously known as penicilliosis), a very important opportunistic systematic mycosis in immunocompromised patients. Different virulence mechanisms in have been proposed and investigated. In the sera of patients with talaromycosis, Mp1 protein (Mp1p), a secretory galactomannoprotein antigen with two tandem ligand-binding domains (Mp1p-LBD1 and Mp1p-LBD2), was found to be abundant. Mp1p-LBD2 was reported to possess a hydrophobic cavity to bind copurified (PLM). It was hypothesized that capturing of lipids from human hosts by expressing a large quantity of Mp1p is a virulence mechanism of It was shown that expression of Mp1p enhanced the intracellular survival of by suppressing proinflammatory responses. Mechanistic study of Mp1p-LBD2 suggested that arachidonic (AA), a precursor of paracrine signaling molecules for regulation of inflammatory responses, is the major physiological target of Mp1p-LBD2. In this study, we use crystallographic and biochemical techniques to further demonstrate that Mp1p-LBD1, the previously unsolved first binding domain of Mp1p, is also a strong AA-binding domain in Mp1p. These studies on Mp1p-LBD1 support the idea that the highly expressed Mp1p is an effective AA-capturing protein. Each Mp1p can bind up to 4 AA molecules. The crystal structure of Mp1p-LBD1-LBD2 has also been solved, showing that both LBDs are likely to function independently with a flexible linker between them. and potentially other pathogens highly expressing and secreting proteins similar to Mp1p can severely disturb host signaling cascades during proinflammatory responses by reducing the availabilities of important paracrine signaling molecules.Copyright © 2019 American Society for Microbiology.

Keyword: fat metabolism

Gibberellin Promotes Cell Growth and Induces Changes in Fatty Biosynthesis and Upregulates Fatty Biosynthetic Genes in Chlorella vulgaris UMT-M1.

Microalgae lipids and oils are potential candidates for renewable biofuels and nutritional inventions. Recent studies from our lab have shown that two plant hormones, auxin and jasmonic , influence microalgae growth and fatty accumulation. Therefore, in this study, a high oil-producing strain Chlorella vulgaris UMT-M1 was selected for hormonal study using gibberellin (GA). Exogenous GA was applied to early stationary culture of C. vulgaris UMT-M1. Results showed that GA gradually increases the cell density of C. vulgaris to up to 42% on days after treatment (DAT)-8 and also capable of delaying the algal senescence. However, the increment in cell density did not enhance the total oil production albeit transient modification of fatty compositions was observed for saturated (SFA) and polyunsaturated (PUFA) fatty acids. This illustrates that GA only promotes cell division and growth but not the oil accumulation. In addition, application of GA in culture medium was shown to promote transient increment of (C16:0) and stearic (C18:0) acids from DAT-4 to DAT-6 and these changes are correlated with the expression of β-ketoacyl ACP synthase I (KAS I) gene.

Keyword: fat metabolism

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the liver of high- and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in liver sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Keyword: fat metabolism

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and diabetes, and (3) dietary SFA type impacts LCPUFA .Published by Elsevier Inc.

Keyword: fat metabolism

The Spatiotemporal Deposition of Lysophosphatidylcholine Within Starch Granules of Maize Endosperm and its Relationships to the Expression of Genes Involved in Endoplasmic Reticulum-Amyloplast Trafficking and Galactolipid Synthesis.

The presence of lipids within starch granules is specific to cereal endosperm starches. These starch lipids are composed of lysophospholipids, especially lysophosphatidylcholine (LysoPC) and free fatty acids that strongly impact the assembly and properties of cereal starches. However, the molecular mechanisms associated with this specific routing have never been investigated. In this study, matrix-assisted laser desorption ionization mass spectrometry imaging revealed decreasing gradients in starch LysoPC concentrations from the periphery to the center of developing maize endosperms. This spatiotemporal deposition of starch LysoPC was similar to that previously observed for endoplasmic reticulum (ER)-synthesized storage proteins, i.e. zeins, suggesting that LysoPC might originate in the ER, as already reported for chloroplasts. Furthermore, a decrease of the palmitate concentration of amyloplast galactolipids was observed during endosperm development, correlated with the preferential trapping of palmitoyl-LysoPC by starch carbohydrates, suggesting a link between LysoPC and galactolipid synthesis. Using microarray, the homologous genes of the Arabidopsis ER-chloroplast trafficking and galactolipid synthesis pathways were also expressed in maize endosperm. These strong similarities suggest that the encoded enzymes and transporters are adapted to managing the differences between chloroplast and amyloplast homeostasis. Altogether, our results led us to propose a model where ER-amyloplast trafficking directs the LysoPC towards one of two routes, the first towards the stroma and starch granules and the other towards galactolipid synthesis.

Keyword: fat metabolism

Hepatocyte-Derived Lipotoxic Extracellular Vesicle Sphingosine 1-Phosphate Induces Macrophage Chemotaxis.

The pathophysiology of non-alcoholic steatohepatitis involves hepatocyte lipotoxicity due to excess saturated free fatty acids and concomitant proinflammatory macrophage effector responses. These include the infiltration of macrophages into hepatic cords in response to incompletely understood stimuli. Stressed hepatocytes release an increased number of extracellular vesicles (EVs), which are known to participate in intercellular signaling and coordination of the behavior of immune cell populations via their cargo. We hypothesized that hepatocyte-derived lipotoxic EVs that are enriched in sphingosine 1-phosphate (S1P) are effectors of macrophage infiltration in the hepatic microenvironment. Lipotoxic EVs were isolated from palmitate treated immortalized mouse hepatocytes and characterized by nanoparticle tracking analysis. Lipotoxic EV sphingolipids were quantified using tandem mass spectrometry. Wildtype and S1P receptor knockout bone marrow-derived macrophages were exposed to lipotoxic EV gradients in a microfluidic gradient generator. Macrophage migration toward EV gradients was captured by time-lapse microscopy and analyzed to determine directional migration. Fluorescence-activated cell sorting along with quantitative PCR and immunohistochemistry were utilized to characterize the cell surface expression of S1P receptor on intrahepatic leukocytes and hepatic expression of S1P receptor, respectively. Palmitate treatment induced the release of EVs. These EVs were enriched in S1P. Palmitate-induced S1P enriched EVs were chemoattractive to macrophages. EV S1P enrichment depended on the activity of sphingosine kinases 1 and 2, such that, pharmacological inhibition of sphingosine kinases 1 and 2 resulted in a significant reduction in EV S1P cargo without affecting the number of EVs released. When exposed to EVs derived from cells treated with palmitate in the presence of a pharmacologic inhibitor of sphingosine kinases 1 and 2, macrophages displayed diminished chemotactic behavior. To determine receptor-ligand specificity, we tested the migration responses of macrophages genetically deleted in the S1P receptor toward lipotoxic EVs. S1P receptor knockout macrophages displayed a marked reduction in their chemotactic responses toward lipotoxic palmitate-induced EVs. Palmitate-induced lipotoxic EVs are enriched in S1P through sphingosine kinases 1 and 2. S1P-enriched EVs activate persistent and directional macrophage chemotaxis mediated by the S1P receptor, a potential signaling axis for macrophage infiltration during hepatic lipotoxicity, and a potential therapeutic target for non-alcoholic steatohepatitis.

Keyword: fat metabolism

is a novel hypothalamic gene upregulated by a high- diet and leptin in mice.

Energy homeostasis is regulated by the hypothalamus but fails when animals are fed a high- diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated , oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic inflammation.

Keyword: fat metabolism

Berberine (BBR) Attenuated (PA)-Induced Lipotoxicity in Human HK-2 Cells by Promoting Peroxisome Proliferator-Activated Receptor α (PPAR-α).

BACKGROUND Berberine (BBR), a natural alkaloid isolated from Coptis chinensis, has frequently been reported as an antidiabetic reagent, partly due to its -lowering activity. Evidence suggests that BBR ameliorates palmitate-induced deposition and apoptosis in renal tubular epithelial cells (TECs), which tracks in tandem with the enhancement of peroxisome proliferator-activated receptor alpha (PPAR-alpha). The study aim was to investigate the roles of BBR in renal lipotoxicity in vitro, and investigate whether PPAR-alpha was the underlying mechanism. MATERIAL AND METHODS Human TECs (HK-2 cells) were injured with (PA), and then treated with BBR, BBR+PPAR-alpha inhibitor (GW6471), and PA+PPAR-alpha agonist (fenofibrate). Endoplasmic reticulum (ER) stress was assessed by measuring the expression of prospective evaluation of radial keratotomy (PERK), C/EBP-homologous protein (CHOP), and 78 kDa glucose-regulated protein (GRP78). was assessed by determining anabolism-associated genes, including fatty synthase (FAS), acetyl-CoA carboxylase (ACC), and lipoprotein lipase (LPL), as well as catabolism-associated gene, including carnitine palmitoyl transferase 1 (CPT1). Inflammatory response of HK-2 cells was evaluated by measuring interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha. Cell apoptosis and protein levels of cleaved-caspase-3 were evaluated. RESULTS PA downregulated PPAR-alpha and induced server lipotoxicity in HK-2 cells by ER stress, increasing deposition, and elevating inflammatory response of HK-2 cells accompanied with inducting cell apoptosis and cleaved-caspase-3, which were obviously reversed by additional treatment of BBR or PPAR-alpha agonist. However, the protective effect of BBR in PA-induced lipotoxicity in HK-2 cells was significantly ameliorated by PPAR-alpha inhibitor. CONCLUSIONS BBR attenuated PA-induced lipotoxicity via the PPAR-alpha pathway.

Keyword: fat metabolism

Acanthoic modulates lipogenesis in nonalcoholic fatty liver disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic fatty liver disease (NAFLD) associated with accumulation by activating Farnesoid X receptor (FXR) and liver X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high- (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, liver index, and liver droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating accumulation and fatty synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Advanced Liver Fibrosis Is Independently Associated with and Insulin Levels in Patients with Non-Alcoholic Fatty Liver Disease.

Changes in occur during the development and progression non-alcoholic fatty liver disease (NAFLD). However, the fatty (FA) profile in red blood cells (RBC) from patients with liver fibrosis remains unexplored. Thus, the goal of this study was to evaluate the fatty profile in RBC, dietary intake and insulin resistance indicators in patients with NAFLD, according to the degree of hepatic fibrosis. Using elastography, patients were classified with ( = 52) and without ( = 37) advanced liver fibrosis. The fatty profile in RBC was analyzed using gas chromatography and the intake was evaluated through a 24-h dietary recall. Subjects with advanced liver fibrosis had higher levels of , stearic and oleic and total monounsaturated fatty (MUFA) and insulin ( < 0.05), and lower levels of elongase very long chain fatty acids protein-6 and the delta-5-desaturase enzymatic activity ( < 0.05). These results suggest a lack of regulation of enzymes related to FA in patients with advanced fibrosis.

Keyword: fat metabolism

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free fatty (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fat metabolism

pH effects on the and fatty acids accumulation in Chlamydomonas reinhardtii.

pH variations influence the delivery of essential nutrients and CO solubility, which impact algae . In this study the microalgal growth and chlorophyll, , and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. content increased 3.0 (14.81% of dry cell weight, dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for ; 17.3, 14.7, and 25.7% for oleic ; and 9.8, 12.1, and 4.6% for linoleic ; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and accumulation.© 2019 American Institute of Chemical Engineers.

Keyword: fat metabolism

d- chiro-Inositol Ameliorates High Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high diet and saturated -treated hepatocytes. DCI attenuated free fatty uptake by the liver via trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: fat metabolism

Fatty acids uptake and oxidation are increased in the liver of rats with adjuvant-induced arthritis.

Severe rheumatoid cachexia is associated with pronounced loss of muscle and mass in patients with advanced rheumatoid arthritis. This condition is associated with dyslipidemia and predisposition to cardiovascular diseases. Circulating levels of triglycerides (TG) and free fatty acids (FFA) have not yet been consistently defined in severe arthritis. Similarly, the of these lipids in the arthritic liver has not yet been clarified. Aiming at filling these gaps this study presents a characterization of the circulating profile and of the fatty acids uptake and in perfused livers of rats with adjuvant-induced arthritis. The levels of TG and total cholesterol were reduced in both serum (10-20%) and liver (20-35%) of arthritic rats. The levels of circulating FFA were 40% higher in arthritic rats, possibly in consequence of cytokine-induced adipose tissue lipolysis. Hepatic uptake and oxidation of and oleic acids was higher in arthritic livers. The phenomenon results possibly from a more oxidized state of the arthritic liver. Indeed, NADPH/NADP and NADH/NAD ratios were 30% lower in arthritic livers, which additionally presented higher activities of the citric cycle driven by both endogenous and exogenous FFA. The lower levels of circulating and hepatic TG possibly are caused by an increased oxidation associated to a reduced synthesis of fatty acids in arthritic livers. These results reveal that the hepatic in arthritic rats presents a strong catabolic tendency, a condition that should contribute to the marked cachexia described for arthritic rats and possibly for the severe rheumatoid arthritis.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

Metabolic changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high- diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: fat metabolism

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic . Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 diabetes animal models. After streptozotocin (STZ) or high- diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

In vivo kinetic study of materno-fetal fatty transfer in obese and normal weight pregnant women.

Placental structure and function can be modified as a result of maternal obesity affecting materno-fetal fatty acids (FA) transport. We report for the first time, in humans and in vivo, the kinetics of placental FA transfer in normo-weight and in normolipemic obese pregnant women using stable isotopes. The administration of different tracer FA with similar behaviour to the mother at different time points allows the collection of kinetic information on materno-fetal transfer of FA despite only one sample of placenta and cord can be collected per subject. Computational modelling showed a good fit to the data when considering all maternal plasma classes but not when based only on non-esterified FA. The novel approach using multiple tracer FA administration combined with computational modelling shows a consistent time course of placental tracer FA and predicted total FA accumulation.We analyse for the first time the in vivo materno-fetal kinetic transfer of fatty acids (FA) labelled with stable isotopes in control and obese (OB) pregnant women. Labelled FA with a similar (stearic : C-SA; : C-PA; oleic : C-OA) were orally administered at -4\xa0h, -8\xa0h and -12\xa0h, respectively prior to elective caesarean section to 10 pregnant women with a body mass index >30 (OB) and 10 with a body mass index in the range 20-25 (NW). Placenta, venous and arterial cord blood were collected obtaining a wide range of FA enrichments. A combined experimental and computational modelling analysis was applied. FA fractional synthesis rate (FSR) in placenta was 11-12%\xa0h . No differences were observed between NW and normo-lipidemic OB. It was not possible to estimate FA FSR in cord blood with this oral bolus dose approach. Computational modelling demonstrated a good fit to the data when all maternal plasma classes were included but not with modelling based only on the non-esterified FA fraction. The estimated materno-fetal C-FA transfer was ∼1%. In conclusion, our approach using multiple C-FA tracers allowed us to estimated FSR in placental/maternal plasma but not in fetal/maternal compartments. Computational modelling showed a consistent time course of placental C-FA transfer and predicted total fetal FA accumulation during the experiment. We conclude that, in addition to non-esterified FA fraction in the maternal circulation, maternal plasma very low-density lipoprotein and other lipoproteins are important contributors to placental FA transfer to the fetus.© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.

Keyword: fat metabolism

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of , Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free fatty (FFA)- and oleic -and high- diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice liver were measured, and the expression levels of genes associated with lipogenesis, fatty oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with fatty oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of , inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: fat metabolism

Unsaturated Fatty Acids Increase the Expression of Hepassocin through a Signal Transducer and Activator of Transcription 3-Dependent Pathway in HepG2 Cells.

Hepassocin (HPS) is a hepatokine that regulates hepatocyte proliferation. It is known that HPS plays an important role in the development of nonalcoholic fatty liver diseases (NAFLD). Fatty acids, such as oleic (OLA), exhibit the ability to activate the signal transducer and activator of transcription-3 (STAT3), and the binding site of STAT3 is found in the promoter region of HPS. However, the regulation of HPS by fatty acids is still obscure. To clarify the regulation of HPS, we detected the expression of HPS by western blots. In addition, a hepatic steatosis cell culture model was established by treatment of different fatty acids, including linoleic (LNA), oleic , , and stearic . The intracellular accumulation was confirmed by oil red O staining. Blocking of STAT3 activity was achieved by the pretreatment of the STAT3 inhibitor, stattic. We found that activation of STAT3 by interleukin-6 (IL-6) was mediated in the regulation of HPS expression. Treatment of unsaturated fatty acids significantly induced intracellular accumulation in HepG2 cells. Moreover, the expressions of HPS were increased in unsaturated fatty -treated HepG2 cells, as compared with saturated fatty -treated groups. Also, the expression of HPS induced by OLA was blocked by the inhibition of STAT3 activity. Furthermore, we found that deletion of HPS by small interfering ribonucleic transfection decreased the protective effect of OLA on cell viability. Taken together, we provided evidence that STAT3 plays an important role in the regulation of OLA-induced HPS expression and the increased HPS may further participate in the development of NAFLD. In addition, the increase of HPS might be involved in the protective effect of OLA on cell viability.© 2018 AOCS.

Keyword: fat metabolism

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty . Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: fat metabolism

Role of the mTOR‑FOXO1 pathway in obesity‑associated renal tubulointerstitial inflammation.

Since obesity is largely responsible for the growing incidence of renal tubulointerstitial inflammation, exploration into the mechanisms of obesity‑associated tubulointerstitial inflammation is essential. Studies have demonstrated that mammalian target of rapamycin\xa0(mTOR) is a crucial molecule in the pathogenesis of renal inflammation, including regulating the expression of inflammatory factors. The purpose of the present study was to further elucidate the role of mTOR in obesity‑associated tubulointerstitial inflammation. In the clinical study, obese and healthy subjects were recruited for physical examination, as well as the collection of blood and urine samples. Further study was performed on a high diet\xa0(HFD)‑induced obese rat model and a cultured human renal tubular epithelial cell line\xa0(HK‑2). The clinical study demonstrated that the participants with obesity had increased serum lipids, creatinine\xa0(Cr), urinary albumin to creatinine ratio\xa0(UACR) and urinary neutrophil gelatinase‑associated lipocalin\xa0(u‑NGAL). Moreover, the level of urinary monocyte chemoattractant protein‑1\xa0(u‑MCP‑1) was increased in the participants with obesity, and it was positively correlated with free fatty \xa0(FFA), UACR and u‑NGAL. In the in\xa0vivo study, the results indicated that the levels of serum lipids, Cr and blood urea nitrogen\xa0(BUN), as well as 24\xa0h urine protein and u‑NGAL, were significantly increased in the HFD‑fed obese rats. In addition, the infiltration of CD68+ cells into the renal interstitial area and the release of interleukin‑1β\xa0(IL‑1β) was observed in the kidneys of obese rats. Meanwhile, the supernatant from HK‑2 cells treated with stimulated THP‑1 monocyte migration. The upregulation of MCP‑1, phosphorylated forkhead boxO1 (p‑FOXO1), and phosphorylated mTOR (p‑mTOR) was observed in\xa0vivo and in\xa0vitro. However, inhibition of mTOR was able to alleviate the above effects. Overall, these results demonstrated that activated mTOR induced FOXO1 phosphorylation, which mediates renal MCP‑1 release, causes tubulointerstitial inflammation and ultimately leads to pathological renal changes and dysfunction. However, inhibition of mTOR may play a renoprotective role during the progression of obesity‑associated tubulointerstitial inflammation.

Keyword: fat metabolism

Cellular toxicity of dietary trans fatty acids and its correlation with ceramide and diglyceride accumulation.

High fatty (FA) levels are deleterious to pancreatic β-cells, largely due to the accumulation of biosynthetic intermediates, such as ceramides and diglycerides, which induce ER stress and apoptosis. Toxicity of palmitate (16:0) and oleate (18:1 cis-Δ) has been widely investigated, while very little data is available on the cell damages caused by elaidate (18:1 trans-Δ) and vaccenate (18:1 trans-Δ), although the potential health effects of these dietary trans fatty acids (TFAs) received great publicity. We compared the effects of these four FAs on cell viability, apoptosis, ER stress, JNK phosphorylation and autophagy as well as on ceramide and diglyceride contents in RINm5F insulinoma cells. Similarly to oleate and unlike palmitate, TFAs reduced cell viability only at higher concentration, and they had mild effects on ER stress, apoptosis and autophagy. Palmitate increased ceramide and diglyceride levels far more than any of the unsaturated fatty acids; however, incorporation of TFAs in ceramides and diglycerides was strikingly more pronounced than that of oleate. This indicates a correlation between the accumulation of intermediates and the severity of cell damage. Our findings reveal important metabolic characteristics of TFAs that might underlie a long term toxicity and hence deserve further investigation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high- diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high- diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the liver, liver/body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high- diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high- diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Dynamics of Individual Fatty Acids in Muscle Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, metabolic rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C to C diet shift experiment and compound-specific δC analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic (LA; 18:2n6) and (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic (20:4n6) and docosahexaenoic (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in energy . The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects energy of birds during migration.

Keyword: fat metabolism

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) . However, whether TMAO aggravates liver steatosis by modulating BA and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA . Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high- diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fat metabolism

Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering hyperlipidemia via regulating the AMPK/SREBP1/PPARα signaling cascade of .

The present study for the first time aims to examine the hypothesis that circulating gamma-glutamyl carboxylated growth arrest specific protein 6 (Gla-Gas6) deficiency may be associated with hyperlipidemia and vitamin K (VK) supplementation may ameliorate the impaired homeostasis via activating Gas6 protein. Subjects with hyperlipidemia (n=22) and age-matched healthy controls (n=19) were included in this study. Results showed that plasma levels of Gla-Gas6 protein and VK were significantly lower in hyperlipidemic subjects compared to control. Moreover, Gla-Gas6 levels were significantly and positively correlated with VK (P=.034, r=0.452) and negatively with triglyceride (P=.022, r=-0.485) and total cholesterol (P=.043, r=-0.435) in hyperlipidemic subjects, which suggest that VK supplementation may have a positive effect in activating Gas6 protein and thereby reducing the aberrant plasma levels. Further studies with high- diet (HFD)-fed animal model of hyperlipidemia demonstrated that VK supplementation (5 μg/kg body weight, 8 weeks) reduced the plasma levels, stimulated both the plasma levels and the hepatic protein expression of Gla-Gas6 protein, and regulated the AMPK/SREBP1/PPARα signaling pathways of hepatic in HFD-fed mice. Moreover, by using (PA, 0.75 mM)-treated both control and GGCX knockdown hepatocytes, this study dissected the direct role of Gla-Gas6 in mediating the positive effect of VK on preventing the PA-induced impaired hepatic via regulating AMPK/SREBP1/PPARα pathways. Combining all, the present study demonstrated the beneficial effect of VK supplementation in preventing the impaired homeostasis via activating VK-dependent Gas6 protein.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 diabetes mellitus (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, fatty acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and liver, gastrocnemius and brown of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: fat metabolism

[Glucagon-like peptide-1 regulates in hepatocytes through Foxo1/3].

Glucagon-like peptide-1 (GLP-1) has been reported to be effective in the treatment of nonalcoholic fatty liver disease (NAFLD). However, the molecular mechanism of GLP-1 on NAFLD is remained unclear. The present study was to detect whether the effect of GLP-1 on triglyceride (TG) content in hepatocytes is dependent on Foxos. HepG2 cells were treated with /oleic for 24 h. The knockdown of Foxo1, Foxo3 was conducted through small interfering RNA (siRNA). Real time PCT (RT-PCR) was used to detect the changes of the SREBP1c and Aco genes in HepG2 cells after Foxo1/3 knockdown. As expected, /oleic increased TG concentration in HepG2 cells [(12.65 ± 1.32) μg/mg vs. (4.32±0.54) μg/mg, 0.05]. Addition of GLP-1 dose (10, 50, 100nmol/L) dependently lowered the TG content and reached plateau at 100 nmol/L of GLP-1 [TG(8.38±1.47) μg/mg]. The GLP-1 effect on TG remained after knocking down either Foxo1 [(9.09±1.34)μg/mg] or Foxo3 [(8.90±1.60) μg/mg] alone, but not when knocking down Foxo1 and Foxo3 (Foxo1/3) together [(14.66±1.77)μg/mg]. Moreover, knocking down Foxo1/3 also abolished GLP-1 effect on SREBP1c and Aco expression. GLP-1 can inhibit the synthesis of TG in hepatocytes depending on Foxo1 and Foxo3. Further studies are needed to explore the specific mechanisms.

Keyword: fat metabolism

Encapsulation of soybean meal with fats enriched in and stearic acids: effects on rumen-undegraded protein and in vitro intestinal digestibility.

coating of soybean meal (SBM) can reduce its protein degradability in the rumen, but the encapsulation of SBM with (PA) and stearic acids (SA) has not yet been investigated, despite both fatty acids are common energy sources in dairy cow diets. This study aimed to evaluate the effects of applying a novel method, using either 400 or 500\xa0g /kg (treatments FL40 and FL50, respectively), which was enriched in PA and SA at different ratios (100:0, 75:25, 50:50, 25:75 and 0:100), on physical and chemical characteristics, ruminal degradability, solubility and in vitro intestinal protein digestibility (IVIPD) of the obtained products. Encapsulation of SBM in resulted in greater mean particle size and lower bulk density and protein solubility than unprotected SBM (USBM). Treatment FL50 resulted in increased (p <\xa00.01) rumen-undegraded protein (RUP) compared to USBM. There were no differences in RUP of SBM when different PA: SA ratios were used. The mean RUP content of treatments FL40 and FL50 (306 and 349\xa0g/kg, respectively) was greater compared to USBM (262\xa0g/kg, p <\xa00.05), but lower than that for a standard heat-treated SBM (431\xa0g/kg). Values of IVIPD did not differ among SBM, heat-treated SBM and FL40 and FL50 samples, all being greater than 97.8%. In conclusion, encapsulation of SBM with fats enriched in PA and SA proved to be effective in reducing protein solubility and increasing RUP without depressing protein digestibility in the intestine. For validation of the method, in vivo research to investigate the effects of these products on the production of dairy cows is warranted.

Keyword: fat metabolism

Myo-inositol alters 13C-labeled fatty in human placental explants.

We postulate that myo-inositol, a proposed intervention for gestational-diabetes, affects transplacental supply to the fetus. We investigated the effect of myo-inositol on fatty- processing in human placental-explants from uncomplicated pregnancies. Explants were incubated with 13C-labeled , 13C-oleic- and 13C-docosahexaenoic- across a range of myo-inositol concentrations for 24 h and 48 h. The incorporation of labeled-fatty-acids into individual lipids was quantified by liquid-chromatography-mass-spectrometry. At 24 h, myo-inositol increased the amount of 13C- and 13C-oleic- labeled lipids (median fold-change relative to control=1). Significant effects were seen with 30 µM myo-inositol (physiological) for 13C--lysophosphatidylcholines (1.26) and 13C--phosphatidylethanolamines (1.17). At 48 h, myo-inositol addition increased 13C-oleic--lipids but decreased 13C- and 13C-docosahexaenoic- lipids. Significant effects were seen with 30 µM myo-inositol for 13C-oleic--phosphatidylcholines (1.25), 13C-oleic--phosphatidylethanolamines (1.37) and 13C-oleic--triacylglycerols (1.32) and with 100 µM myo-inositol for 13C-docosahexaenoic--triacylglycerols (0.78). Lipids labeled with the same 13C-fatty- showed similar responses when tested at the same time-point, suggesting myo-inositol alters upstream processes such as fatty- uptake or activation. Myo-inositol supplementation may alter placental physiology with unknown clinical consequences.

Keyword: fat metabolism

Polysaccharides Prevent -Evoked Apoptosis and Autophagy in Intestinal Porcine Epithelial Cell Line via Restoration of Mitochondrial Function and Regulation of MAPK and AMPK/Akt/mTOR Signaling Pathway.

polysaccharide (GLP) extracted from (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high- constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.

Keyword: fat metabolism

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting lipogenesis and inflammation.

Non-alcoholic fatty liver disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), fatty synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fat metabolism

Enhanced Fatty Scavenging and Glycerophospholipid Accompany Melanocyte Neoplasia Progression in Zebrafish.

Alterations in in cancer cells impact cell structure, signaling, and energy , making a potential diagnostic marker and therapeutic target. In this study, we combined PET, desorption electrospray ionization-mass spectrometry (DESI-MS), nonimaging MS, and transcriptomic analyses to interrogate changes in in a transgenic zebrafish model of oncogenic RAS-driven melanocyte neoplasia progression. Exogenous fatty uptake was detected in melanoma tumor nodules by PET using the surrogate tracer 14(R,S)-18F-fluoro-6-thia-heptadecanoic ([18F]-FTHA), consistent with upregulation of genes associated with fatty uptake found through microarray analysis. DESI-MS imaging revealed that FTHA uptake in tumors was heterogeneous. Transcriptome and lipidome analyses further highlighted dysregulation of glycerophospholipid pathways in melanoma tumor nodules, including increased abundance of phosphatidyl ethanolamine and phosphatidyl choline species, corroborated by DESI-MS, which again revealed heterogeneous phospholipid composition in tumors. Overexpression of the gene encoding lipoprotein lipase (LPL), which was upregulated in zebrafish melanocyte tumor nodules and expressed in the majority of human melanomas, accelerated progression of oncogenic RAS-driven melanocyte neoplasia in zebrafish. Depletion or antagonism of LPL suppressed human melanoma cell growth; this required simultaneous fatty synthase (FASN) inhibition when FASN expression was also elevated. Collectively, our findings implicate fatty acquisition as a possible therapeutic target in melanoma, and the methods we developed for monitoring fatty uptake have potential for diagnosis, patient stratification, and monitoring pharmacologic response. SIGNIFICANCE: These findings demonstrate the translational potential of monitoring fatty uptake and identify lipoprotein lipase as a potential therapeutic target in melanoma.©2019 American Association for Cancer Research.

Keyword: fat metabolism

Resveratrol Maintains Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid metabolic disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive oxygen species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular metabolic imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of metabolic disorders relevant to the circadian clock.

Keyword: fat metabolism

Diosgenin ameliorates -induced accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic accumulation. Many studies have suggested that overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy , but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on , glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, consumption and synthesis in LO2 cells was investigated.The results indicated that induced obvious accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: fat metabolism

Vascular endothelial growth factor B inhibits accumulation in C2C12 myotubes incubated with fatty acids.

To investigate (1) the effect of vascular endothelial growth factor B (VEGFB) on accumulation and the alteration of fatty acids and fatty -related enzymes in C2C12 myotubes incubated with fatty acids and (2) the regulatory effect of VEGFB on skeletal muscle . Mouse C2C12 myotubes were incubated with oleic (OA) and (PA), and differentiated mature C2C12 myotubes were treated with VEGFB. Oil-red O staining, BODIPY staining and cell triglycerides (TG) content were examined. Total RNA was isolated, and real-time PCR analysis was performed. Treatment with 100\u2009μM OA and 50\u2009μM PA induced droplet accumulation and increased TG content (\u2009<\u2009.01), and 100\u2009ng/mL VEGFB reduced droplet accumulation and decreased TG content (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of fatty transport protein 1 (FATP1) (\u2009<\u2009.01) and FATP4 (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of adipose TG lipase and hormone-sensitive lipase (\u2009<\u2009.01) as well as carnitine palmitoyltransferase I (\u2009<\u2009.01), peroxisome proliferator-activated receptor-γ coactivator-1α (\u2009<\u2009.01), acyl-coa dehydrogenase very long chain (\u2009<\u2009.05), acyl-coa synthetase long-chain family member 1 (\u2009<\u2009.01), peroxisomal acyl-coenzyme A oxidase 1 (\u2009<\u2009.05), and mitochondrial uncoupling protein 3 (\u2009<\u2009.01). VEGFB enhanced FATP1and FATP4 expression, promoted C2C12 myotube fatty oxidation and TG decomposition, and inhibited C2C12 myotube fatty re-esterification, thus inhibiting accumulation in C2C12 myotubes incubated with fatty acids.

Keyword: fat metabolism

Mitochondrial Metabolic Reprogramming by CD36 Signaling Drives Macrophage Inflammatory Responses.

A hallmark of chronic inflammatory disorders is persistence of pro-inflammatory macrophages in diseased tissues. In atherosclerosis this is associated with dyslipidemia and oxidative stress, but mechanisms linking these phenomena to macrophage activation remain incompletely understood. To investigate mechanisms linking dyslipidemia, oxidative stress and macrophage activation through modulation of immunemetabolism, and to explore therapeutic potential targeting specific metabolic pathways. Using a combination of biochemical, immunological, and ex vivo cell metabolic studies, we report that CD36 mediates a mitochondrial metabolic switch from oxidative phosphorylation to superoxide production in response to its ligand, oxLDL. Mitochondrial-specific inhibition of superoxide inhibited oxLDL-induced NF-κB activation and inflammatory cytokine generation. RNAseq, flow cytometry, H-labeled uptake, lipidomic analysis, confocal and EM imaging, and functional energetics revealed that oxLDL upregulated effectors of long-chain fatty (FA) uptake and mitochondrial import, while downregulating FA oxidation and inhibiting ATP5A, an electron transport chain (ETC) component. The combined effect is long-chain FA accumulation, alteration of mitochondrial structure and function, repurposing of the ETC to superoxide production, and NF-κB activation. Apoe null mice challenged with high diet showed similar metabolic changes in circulating Ly6C monocytes and peritoneal macrophages, along with increased CD36 expression. Moreover, mitochondrial ROS was positively correlated with CD36 expression in aortic lesional macrophages. These findings reveal that oxLDL/CD36 signaling in macrophages links dys-regulated FA to oxidative stress from the mitochondria, which drives chronic inflammation. Thus, targeting to CD36 and its downstream effectors may serve as potential new strategies against chronic inflammatory diseases such as atherosclerosis.

Keyword: fat metabolism

Soluble epoxide hydrolase derived mediators are elevated in bronchoalveolar lavage fluid from patients with sarcoidosis: a cross-sectional study.

Sarcoidosis is a systemic inflammatory multi-organ disease almost always affecting the lungs. The etiology remains unknown, but the hallmark of sarcoidosis is formation of non-caseating epithelioid cells granulomas in involved organs. In Scandinavia, >\u200930% of sarcoidosis patients have Löfgren\'s syndrome (LS), an acute disease onset mostly indicating a favorable prognosis. The impact of dysregulation of mediators, which has been investigated in other inflammatory disorders, is still unknown.Using three different liquid chromatography coupled to tandem mass spectrometry targeted platforms (LC-MS/MS), we quantified a broad suite of mediators including eicosanoids, sphingolipids and endocannabinoids in bronchoalveolar lavage (BAL) fluid from pulmonary sarcoidosis patients (n\u2009=\u200941) and healthy controls (n\u2009=\u200916).A total of 47 mediators were consistently detected in BAL fluid of patients and controls. After false discovery rate adjustment, two products of the soluble epoxide hydrolase (sEH) enzyme, 11,12-dihydroxyeicosa-5,8,14-trienoic (11,12-DiHETrE, p\u2009=\u20094.4E-5, q\u2009=\u20091.2E-3, median fold change\u2009=\u20096.0) and its regioisomer 14,15-dihydroxyeicosa-5,8,11-trienoic (14,15-DiHETrE, p\u2009=\u20093.6E-3, q\u2009=\u20093.2E-2, median fold change\u2009=\u20091.8) increased in patients with sarcoidosis. Additional shifts were observed in sphingolipid , with a significant increase in -derived sphingomyelin (SM16:0, p\u2009=\u20091.3E-3, q\u2009=\u20091.7E-2, median fold change\u2009=\u20091.3). No associations were found between these 3 mediators and LS, whereas levels of SM 16:0 and 11,12-DiHETrE associated with radiological stage (p\u2009<\u20090.05), and levels of 14,15-DiHETrE were associated with the BAL fluid CD4/CD8 ratio.These observed shifts in mediators provide new insights into the pathobiology of sarcoidosis and in particular highlight the sEH pathway to be dysregulated in disease.

Keyword: fat metabolism

Energy and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of energy from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body energy and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net energy and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible energy basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty constituents are , oleic, and linoleic ), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine energy and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross energy intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated energy retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk yield and milk energy output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable energy intake, increased milk yield and milk energy output, increased plasma triacylglycerides and long-chain fatty concentrations, and had no effect on energy retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable energy intake, and total milk, , protein, and lactose yields. Milk energy output increased at the HMP level, and protein level did not affect total energy retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted energy retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra energy intake into milk and had no effect on milk N efficiency.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Apoptotic germ cells regulate Sertoli cell storage and fatty oxidation

The presence of droplets (LDs) and the utilization of fatty acids (FAs) as a source of energy are Sertoli cell (SC) putative characteristics. It is well known that SCs can phagocyte and degrade apoptotic germ cells (AGCs) resulting in increasing content and ATP levels. A relationship between the regulation of storage and of oxidation in SC might be envisaged. The aim of this study was to analyze whether AGCs and FAs are able to simultaneously regulate molecular mechanisms involved in storage and in FA oxidation in SC. The experimental model utilized in this study consisted in SC cultures obtained from 20-day-old rats that were co-cultured with AGC or treated with (PA, 500 μM) for 24 and 48 h. AGC and PA increase LD, triacylglycerol (TAG) content and mRNA levels of Plin1, Plin2, Plin3 (proteins involved in TAG storage). Simultaneously, AGC and PA rise the extent of FA oxidation and mRNA levels of Cpt1 and Lcad (proteins involved in FA degradation). Results also show that peroxisome proliferator-activated receptor (PPAR) transcriptional activity, transcription factor which participate in regulation, increases by AGC and PA treatment in SC. Additionally, the presence of a PPARg antagonist decreases the upregulation of LD content and Plin1 expression. Similarly, the presence of a PPARb/d antagonist reduces the increase in FA oxidation and Cpt1 mRNA levels. Altogether these results suggest that AGC and FA, which probably generate PPAR ligands, regulate storage and fatty utilization, contributing to the energy homeostasis in the seminiferous tubules.

Keyword: fat metabolism

Chronic intake of moderate -enriched diet induces fatty liver and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate -enriched diet on hepatic accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of -enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic fatty liver disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), cirrhosis, and cancer. The role of different types of fatty acids in driving the early progression of NAFL to NASH is not understood. overload causing lipotoxicity and inflammation has been considered as an essential pathogenic factor. To correlate the profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced inflammation and cell death, whereas C16:1 induced significant droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific fatty acids that affect and cause lipotoxicity or droplet formation. It indicates that not only saturation or monounsaturation of fatty acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of and the heterogeneous development of NAFLD.

Keyword: fat metabolism

production by Lipomyces starkeyi using sap squeezed from felled old oil palm trunks.

The ability of oleaginous yeast Lipomyces starkeyi to efficiently produce lipids when cultivated on sap extracted from felled oil palm trunk (OPT) as a novel inexpensive renewable carbon source was evaluated. OPT sap was found to contain approximately 98\xa0g/L glucose and 32\xa0g/L fructose. Batch fermentations were performed using three different OPT sap medium conditions: regular sap, enriched sap, and enriched sap at pH 5.0. Under all sap medium conditions, the cell biomass and production achieved were approximately 30\xa0g/L and 60% (w/w), respectively. L.\xa0starkeyi tolerated acidified medium (initial pH ≈ 3) and produced considerable amounts of ethanol as well as xylitol as by-products. The fatty profile of L.\xa0starkeyi was remarkably similar to that of palm oil, one of the most common vegetable oil feedstock used in biodiesel production with oleic as the major fatty followed by , stearic and linoleic acids.Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high- diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: fat metabolism

Fatty Profile and Cardiometabolic Markers in Relation with Diet Type and Omega-3 Supplementation in Spanish Vegetarians.

Plant-based diets are becoming increasingly popular, and scientific information concerning the nutritional status in this population is needed. This study determined the fatty profile of Spanish lacto-ovo vegetarians (LO-vegetarians) and vegans. Participants were 104 healthy adults, LO-vegetarians ( = 49) and vegans ( = 55). Lifestyle habits and consumption of food and omega-3 supplements were estimated by questionnaires. BMI, blood pressure, and abdominal and body were determined. Serum was collected to analyze fatty acids, glucose, lipids, homocysteine, insulin, and leptin. Volunteers were classified according to serum omega-6 to omega-3 (-6/-3) ratio into three groups: -6/-3 < 10, -6/-3 ≥ 10 to 20, and -6/-3 > 20. Results showed low cardiovascular risk and high insulin sensitivity with negligible differences between diet types. Linoleic (C18:2-6) was the major serum fatty , followed by oleic (C18:1-9) and (C16:0) acids. In contrast, serum eicosapentaenoic (EPA, C20:5-3) and docosahexaenoic (DHA, C22:6-3) were (median, interquartile range) 0.27, 0.18% and 1.59, and 0.93%, respectively. Users of -3 supplements (<10% of total vegetarians) had significantly higher EPA than non-users, while frequent consumption of flax-seeds was associated with increased α-linolenic (C18:3-3). However, neither -3 supplementation nor food consumption affected DHA levels in this vegetarian population.

Keyword: fat metabolism

Berberine attenuates sodium palmitate-induced accumulation, oxidative stress and apoptosis in grass carp(Ctenopharyngodon idella)hepatocyte in vitro.

The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in accumulation-hepatocytes induced by sodium palmitate in vitro. The accumulation-hepatocytes (induced by 0.5\u202fmM sodium palmitate for 24\u202fh) were treated with 5\u202fμM berberine for 12\u202fh. Then, the Cell viability, intracellular triglyceride (TG) content, peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from mitochondria into the cytoplasm. Beberine could improve intracellular droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.Copyright © 2019. Published by Elsevier Ltd.

Keyword: fat metabolism

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 diabetes. Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and . Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: fat metabolism

lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑ diet (HFD)‑fed mice to verify the role of MEG3 in hepatic insulin resistance in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 diabetes mellitus.

Keyword: fat metabolism

Effects of timing of supplementation during early lactation on nutrient digestibility, energy balance, and of dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation during early lactation on nutrient digestibility, energy intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible energy intake by 3.9 Mcal/d, metabolizable energy intake by 3.5 Mcal/d, and net energy for lactation intake by 2.5 Mcal/d. The PA diet also increased milk energy output, negative energy balance, and plasma nonesterified fatty concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for energy balance due to cows receiving the PA treatment being in a greater negative energy balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased energy intake and milk energy output and did not affect energy balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and energy intake compared with a control diet containing no supplemental . Feeding C16:0 markedly increased milk energy output in both the FR and PK periods but increased negative energy balance only in the FR period.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Glycerolipid Composition of the Red Macroalga and Comparison to the Closely Related Producing Different Types of Eicosanoids.

The red macroalga is a well-known producer of eicosanoids such as hydroxyeicosatetraenoic acids, but the alga produces almost no prostaglandins, unlike the closely related . This indicates that the related two algae would have different enzyme systems or substrate composition. To carry out more in-depth discussions on the metabolic pathway of eicosanoids between the two algae, we investigated the characteristics of glycerolipids, which are the substrates of eicosanoids production, of and compared them to the reported values of . In , monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylcholine (PC) were the major classes and accounted for 44.4% of the total extract. The predominant fatty acids were arachidonic (20:4n-6), an eicosanoids precursor, and (16:0). The 20:4n-6 content was extremely high in MGDG and PC (>70%), and the 16:0 content was extremely high in DGDG and SQDG (>40%). A chiral-phase HPLC analysis showed that fatty acids were esterified at the -1 and -2 positions of those lipids. The glycerolipid molecular species were determined by reversed-phase HPLC⁻ESI⁻MS analysis. The main glycerolipid molecular species were 20:4n-6/20:4n-6 (-1/-2) for MGDG (63.8%) and PC (48.2%), 20:4n-6/16:0 for DGDG (71.1%) and SQDG (29.4%). These characteristics of were almost the same as those of . Hence, the differences of the eicosanoids producing ability between the two algae would not be due to the difference of substrate composition but the difference of enzyme system.

Keyword: fat metabolism

Electric pulse stimulation inhibited accumulation on C2C12 myotubes incubated with oleic and .

To investigate the effect of electrical pulse stimulation (EPS) on accumulation and alteration of fatty -related enzymes in C2C12 myotubes incubated with fatty acids. Mouse C2C12 myotubes were incubated with oleic and , and differentiated C2C12 myotubes were treated with EPS, oil-red O (ORO), BODIPY staining and triglyceride (TG) content were examined. Total RNA was isolated, and real-time polymerase chain reaction analysis was performed. (1) EPS decreased TG content (\u2009<\u2009.01). (2) EPS significantly induced the mRNA expression of FAD/CD36 (\u2009<\u2009.05), FATP4 (\u2009<\u2009.001), FABP1 (\u2009<\u2009.01) and FABP5 (\u2009<\u2009.01). (3) EPS significantly inhibited the mRNA expression of fatty synthase (\u2009<\u2009.01). (4) Adipose triglyceride lipase and hormone-sensitive lipase expression were significantly elevated (\u2009<\u2009.001), and induced the mRNA expression of CPT1 (\u2009<\u2009.01), ACOX1 (\u2009<\u2009.05), UCP3 (\u2009<\u2009.05) and PPARα (\u2009<\u2009.001) after EPS. EPS reduced droplet accumulation; enhanced CD36, FATP4, FABP1 and FABP5 expression; inhibited C2C12 myotube fatty re-esterification; and promoted fatty oxidation in C2C12 myotubes.

Keyword: fat metabolism

Optimization of Synechococcus sp. VDW Cultivation with Artificially Prepared Shrimp Wastewater for Ammonium Removal and Its Potential for Use As a Biofuel Feedstock.

To investigate the potential of application of marine cyanobacterium for concurrent biomass production and ammonium removal, Synechococcus sp. VDW was cultured under different conditions in medium containing varying concentrations of NHCl. Response surface methodology (RSM) was then used to build a predictive model of the combined effects of independent variables (pH, inoculum size, ammonium concentration). At the optimum conditions of initial pH 7.4, inoculum size 0.17 (OD730) and ammonium concentration 10.5 mg L, the maximum ammonium removal and biomass productivity were about 95% and 34 mg Ld, respectively, after seven days of cultivation. The result of fatty methyl ester (FAME) analysis showed that the major fatty acids were (C16:0), linoleic (C18:2 n6 cis), palmitoleic (C16:1) and oleic (C18:1 n9 cis), which accounted for more than 80% weight of total fatty acids. Further, analysis of neutral accumulation using flow cytometry revealed that the mean of the fluorescence intensity increased under optimal conditions. These results indicate that Synechococcus sp. VDW has the potential for use for concurrent water treatment and production of biomass that can be applied as biofuel feedstock.

Keyword: fat metabolism

Effects of timing of supplementation on production responses of early-lactation dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental (CON) or a diet supplemented with C16:0 (, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% -corrected milk by 5.30 kg/d, yield of energy-corrected milk (ECM) by 4.70 kg/d, milk content by 0.41% units, milk yield by 280 g/d, and protein yield by 100 g/d. The increase in milk associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased body weight (BW) by 21 kg and body condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% -corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk content by 0.22% units, milk yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk yield due to feeding PA during the PK period increasing milk yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High--Diet-Induced Cardiac Dysfunction and Insulin Resistance by Elevating Tafazzin in Mouse Hearts.

High (HF)-diet-induced insulin resistance is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac insulin signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs insulin signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the mitochondria. Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores insulin sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced insulin resistance further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to insulin resistance via mediating Drp-1 translocation to the mitochondria, and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates insulin resistance.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fat metabolism

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high- diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high- diet (HFD; 60 kcal% ) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic .LLE significantly decreased the intracellular accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fat metabolism

Effects of Chlorpyrifos on Cholinesterase and Serine Lipase Activities and in Brains of Rainbow Trout (Oncorhynchus mykiss).

Chlorpyrifos is an organophosphorus insecticide that elicits acute toxicity through inhibition of acetylcholinesterase (AChE), leading to acetylcholine accumulation and prolonged stimulation of cholinergic receptors throughout the central and peripheral nervous systems. Previous studies have indicated that neurodevelopment may also be impaired through alternative pathways, including reduction of cAMP catalyzed downstream events. The upstream initiating events that underlie non-cholinergic neurological actions of chlorpyrifos and other organophosphorus compounds remain unclear. To investigate the potential role of disruption of fatty signaling as a mechanism of toxicity, and fatty profiles were examined to identify alterations that may play a critical role in upstream signaling in the CNS. Juvenile rainbow trout were treated for 7 days with nominal chlorpyrifos concentrations previously reported to diminish olfactory responses (10, 20, and 40\u2009μg/L). While lethality was noted higher doses, measured chlorpyrifos concentrations of 1.38\u2009μg/L (nominal concentration 10\u2009μg/L) significantly reduced the activity of AChE and two serine lipases, monoacylglycerol lipase and fatty amide hydrolase in the brain. Reductions in lysophosphatidylethanolamines (16:0; 18:0, 18:1, and 22:6) derived from the phosphatidylethanolamines and free fatty acids ( acid16:0; Linolenic acid18:3; Eicosadienoic 20:2; Arachidonic 20:4; and Docosahexaenoic 22:6) were also noted, suggesting that chlorpyrifos inhibited the of selected phospholipid signaling precursors at sublethal concentrations. These results indicate that in addition to AChE inhibition, environmentally relevant chlorpyrifos exposure alters serine lipase activity and metabolites in the trout brain, which may compromise neuronal signaling and impact neurobehavioral responses in aquatic animals.© The Author(s) 2019. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Keyword: fat metabolism

Hexadecenoic Fatty Positional Isomers and De Novo PUFA Synthesis in Colon Cancer Cells.

involves delta-9 and delta-6 desaturase enzymes forming palmitoleic (9-16:1; -7 series) and sapienic (6-16:1; -10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty (MUFA) isomers are under development. Furthermore, sapienic can bring to the de novo synthesis of the -10 polyunsaturated fatty (PUFA) sebaleic (5,8-18:2), but such transformations in cancer cells are not known. The model of Caco-2 cell line was used to monitor sapienic supplementation (150 and 300 μM) and provide evidence of the formation of -10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and acids evidenced that remodelling was influenced by the type of fatty and positional isomer, with an increase of 8-18:1, -10 PUFA and a decrease of saturated fats in case of sapienic . Cholesteryl esters were formed only in cases with sapienic . Sapienic was the less toxic among the tested fatty acids, showing the highest ECs and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic increases the distribution of fluid regions, probably connected with the formation of 8-18:1 and the -10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and cancer research.

Keyword: fat metabolism

Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

CHP1 Regulates Compartmentalized Glycerolipid Synthesis by Activating GPAT4.

Cells require a constant supply of fatty acids to survive and proliferate. Fatty acids incorporate into membrane and storage glycerolipids through a series of endoplasmic reticulum (ER) enzymes, but how these enzymes are regulated is not well understood. Here, using a combination of CRISPR-based genetic screens and unbiased lipidomics, we identified calcineurin B homologous protein 1 (CHP1) as a major regulator of ER glycerolipid synthesis. Loss of\xa0CHP1 severely reduces fatty incorporation and storage in mammalian cells and invertebrates. Mechanistically, CHP1 binds and activates GPAT4, which catalyzes the initial rate-limiting step in glycerolipid synthesis. GPAT4 activity requires CHP1 to be N-myristoylated, forming a key molecular interface between the two proteins. Interestingly, upon CHP1 loss, the peroxisomal enzyme, GNPAT, partially compensates for the loss of ER synthesis, enabling cell proliferation. Thus, our work identifies a conserved regulator of glycerolipid and reveals plasticity in synthesis of proliferating cells.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Berberine Protects Glomerular Podocytes via Inhibiting Drp1-Mediated Mitochondrial Fission and Dysfunction.

Elevated levels of plasma free fatty (FFA) and disturbed mitochondrial dynamics play crucial roles in the pathogenesis of diabetic kidney disease (DKD). However, the mechanisms by which FFA leads to mitochondrial damage in glomerular podocytes of DKD and the effects of Berberine (BBR) on podocytes are not fully understood. : Using the db/db diabetic mice model and cultured mouse podocytes, we investigated the molecular mechanism of FFA-induced disturbance of mitochondrial dynamics in podocytes and testified the effects of BBR on regulating mitochondrial dysfunction, podocyte apoptosis and glomerulopathy in the progression of DKD. : Intragastric administration of BBR for 8 weeks in db/db mice significantly reversed glucose and disorders, podocyte damage, basement membrane thickening, mesangial expansion and glomerulosclerosis. BBR strongly inhibited podocyte apoptosis, increased reactive oxygen species (ROS) generation, mitochondrial fragmentation and dysfunction both and . Mechanistically, BBR could stabilize mitochondrial morphology in podocytes via abolishing (PA)-induced activation of dynamin-related protein 1 (Drp1). : Our study demonstrated for the first time that BBR may have a previously unrecognized role in protecting glomerulus and podocytes via positively regulating Drp1-mediated mitochondrial dynamics. It might serve as a novel therapeutic drug for the treatment of DKD.

Keyword: fat metabolism

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

-Rich High Diet Exacerbates Experimental Pulmonary Fibrosis by Modulating Endoplasmic Reticulum Stress.

The impact of lipotoxicity on the development of lung fibrosis is unclear. Saturated fatty acids such as (PA) activate endoplasmic reticulum (ER) stress, a cellular stress response associated with the development of idiopathic pulmonary fibrosis (IPF). We tested the hypothesis that PA increases susceptibility to lung epithelial cell death and experimental fibrosis by modulating ER stress. Total liquid chromatography and mass spectrometry were used to measure fatty content in IPF lungs. Wild-type mice were fed a high diet (HFD) rich in PA or a standard diet (SD) and subjected to bleomycin-induced lung injury. Lung fibrosis was determined by hydroxyproline content. Mouse lung epithelial cells were treated with PA. ER stress and cell death were assessed by Western blotting, TUNEL staining and cell viability assays. IPF lungs had a higher level of PA compared to controls. Bleomycin-exposed mice fed a HFD had significantly increased pulmonary fibrosis associated with increased cell death and ER stress compared to those fed a SD. PA increased apoptosis and activation of the unfolded protein response in lung epithelial cells. This was attenuated by genetic deletion and chemical inhibition of CD36, a fatty transporter. In conclusion, consumption of a HFD rich in saturated increases susceptibility to lung fibrosis and ER stress and PA mediates lung epithelial cell death and ER stress via CD36. These findings demonstrate that lipotoxicity may have a significant impact on the development of lung injury and fibrosis by enhancing pro-death ER stress pathways.

Keyword: fat metabolism

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

Keyword: fat metabolism

Fatty levels alterations in THP-1 macrophages cultured with lead (Pb).

As cardiovascular events are one of the main causes of death in developed countries, each factor potentially increasing the risk of cardiovascular disease deserves special attention. One such factor is the potentially atherogenic effect of lead (Pb) on , and is significant in view of the still considerable Pb environmental pollution and the non-degradability of Pb compounds.Analysis of saturated fatty acids (SFA) (caprylic (C8:0), decanoic (C10:0), lauric (C12:0), tridecanoic (C13:0), myristic (C14:0), pentadecanoic (C15:0), (C16:0), heptadecanoic (C17:0), stearic (C18:0), and behenic (C22:0)), monounsaturated fatty (MUFA) (palmitoleic (C16:1), oleic (18:1w9), trans-vaccenic (C18:1 trans11)), and polyunsaturated fatty (PUFA) (linoleic (C18:2n6), gamma-linolenic (C18:3n6), arachidonic (C20:4n6)), was conducted by gas chromatography. Analysis of stearoyl-CoA desaturase (SCD), fatty desaturase 1 (FADS1) and fatty desaturase 2 (FADS2) expression was performed using qRT-PCR. Oxidative stress intensity (malondialdehyde - MDA concentration) was measured using spectrophotometric method. Intracellular generation of reactive oxygen species (ROS) in macrophages was visualized by fluorescence microscopy and quantitatively measured by plate reader.Pb caused quantitative alterations in FAs profile in macrophages; the effect was Pb-concentration dependent and selective (i.e. concerned only selected FAs). In general, the effect of Pb was biphasic, with Pb levels of 1.25\u2009μg/dL and 2.5\u2009μg/dL being stimulatory, and 10\u2009μg/dL being inhibitory on concentrations of selected FAs. The most potent Pb concentration, resulting in increase in levels of 9 FAs, was 2.5\u2009μg/dL, the Pb-level corresponding to the mean blood Pb concentrations of people living in urban areas not contaminated by Pb. Pb was found to exert similar, biphasic effect on the expression of FADS1. However, Pb decreased, in a concentration-dependent manner, the expression of SCD and FADS2. Pb significantly increased MDA and ROS concentration in macrophages.Environmental Pb exposure might be a risk factor resulting in alterations in FAs levels, oxidative stress and increased MDA concentration in macrophages, which might lead to the formation of foam cells and to inflammatory reactions.Copyright © 2019 Elsevier GmbH. All rights reserved.

Keyword: fat metabolism

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving , antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis (NASH) is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high high fructose (HFHF) diet fed C57BL/6J mice, used as model of NASH. The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, mitochondria are intracellular source of reactive oxygen species (ROS) and important sites of , we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti-NASH potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Mutation of IDH1 aggravates the fatty ‑induced oxidative stress in HCT116 cells by affecting the mitochondrial respiratory chain.

Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the metabolic reprogramming of cancer cells; however their functions in remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic (OA) or (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive oxygen species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of mitochondria, immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty ‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.

Keyword: fat metabolism

Apigenin reduces the excessive accumulation of lipids induced by via the AMPK signaling pathway in HepG2 cells.

In recent years, increasing attention has been paid to diseases caused by excessive accumulation of lipids in the liver with therapeutic agents derived from natural products offering an alternative treatment to conventional therapies. Among these therapeutic agents, apigenin, a natural flavonoid, has been proven to exert various beneficial biological effects. In the present study, the antiadipogenic effects of apigenin in HepG2 cells was investigated. It was demonstrated that the treatment of cells with different concentrations of apigenin for 24 h significantly decreased the -induced increases in total cholesterol (TC) and triglyceride (TG) levels as well as intracellular accumulation. In addition, apigenin increased the phosphorylated-AMP-activated protein kinase (AMPK) levels but decreased the expression levels of 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding protein (SREBP)-1, fatty synthase, and SREBP-2 in a concentration-dependent manner. The present findings suggested that apigenin might improve by activating the AMPK/SREBP pathway to reduce accumulation in the liver.Copyright: © Lu et al.

Keyword: fat metabolism

[Comparison of effects of oleic and on deposition and mTOR / S6K1 / SREBP-1c pathway in HepG2 cells].

To explore the effects of oleic and on deposition and mTOR/S6K1/SREBP-1c pathways in HepG2 cells. The model of steatosis was established with induction of oleic and and was intervened by rapamycin. The changes in droplets were observed after staining the cells with oil Red O. Intracellular triglyceride (TG) contents in cells were measured by TG kit. mTOR, S6K1, and SREBP-1c mRNA expression levels were detected using QRT-PCR. Western blot was used to determine protein expression levels of mTOR, S6K1 and SREBP-1c. Both fatty acids increased droplets in HepG2 cells. Fatty degeneration with elevated TG occurred with significant changes in oleic group lipids. Rapamycin alleviated deposition caused by oleic and and inhibited their induction of increased expression of mTOR, S6K1, and SREBP-1c. QRT-PCR and Western blot results showed that mRNA and protein expressions of mTOR, S6K1, and SREBP-1c in oleic and group were significantly higher than the control group ( < 0.05). The increase was more pronounced in the group ( < 0.05); however, after rapamycin intervention, the expression of mRNA and protein in the three groups were significantly lower ( < 0.05), and the change in group was more pronounced ( < 0.05). Oleic and can induce deposition in HepG2 cells and increase expression of every component of mTOR/S6K1/SREBP-1c pathway; however, Oleic -induced deposition is more pronounced, and the mTOR, S6K1, and SREBP-1c pathway change is more obvious in . Rapamycin has high potent inhibitory effect on -induced deposition. These results specify that synthesis involved in the mTOR/S6K1/SREBP-1c pathways are mainly associated to in HepG2 cells, whereas other signaling pathway may mediate oleic -induced synthesis.

Keyword: fat metabolism

The effect of saturated and unsaturated acids on the production of outer membrane vesicles from and .

The aim of present study is to investigate the effect of acids on the outer membrane vesicles (OMVs) produced by spp. spp. is the important member of Gut microbiota that employ OMVs production for interact with host. Besides, dietary acids could influence on determination of gut microbiota composition and immune response. In this regard, we evaluated the effect of acids on the growth and OMVs production of and . and were grown on BHI broth with and without and palmitoleic acids as saturated and unsaturated acids, respectively. OMVs were extracted using multiple centrifugation and tris-ethylene diamine tetra acetic (EDTA)-Sodium deoxy cholate buffers. Physicochemical properties of OMVs were detected by electron microscopy (SEM), Bradford Coomassie brilliant blue assay and SDS-PAGE. Data were analyzed with One-way ANOVA using SPSS.The growths of both were significantly increased by . Nevertheless, palmitoleic had no significant effect on them. significantly decreased and increased the production of OMVs at low and high concentration, respectively. However, the production of OMVs was not significantly affected by . Although palmitoleic had a significant decreasing effect on the production of OMVs, it significantly increased the production of OMVs at low concentration.In conclusion we reported that had a stimulatory effect on the growth of and and had a dose dependent effect on the production of OMVs. Also producing of OMVs was affected by palmitoleic in a dose dependent manner.

Keyword: fatty liver

Phytochemistry, Traditional Uses and Pharmacological Profile of Rose Hip: A Review.

The fruit of genus Rosa, known as "rose hip", is frequently used in different traditional medicines. Rose hips have long been used to treat kidney stones, gastroenteric ailments, hypertension and respiratory problems such as bronchitis, cough and cold.This review is focused on the ethnopharmacological uses of rose hip as well as phytochemical and pharmacological aspects.Ethno-medical uses of rose hip have been recorded in many countries since a long time. Approximately, 129 chemical compounds have been isolated and identified from rose hip. This fruit contains some major active components such as flavonoids, tannins, anthocyanin, phenolic compounds, oil, organic acids and inorganic compounds. Scientific studies have suggested a wide range of pharmacological activities for rose hip including antioxidant, anti-inflammatory, anti-obesity, anti-cancer, hepatoprotective, nephroprotective, cardioprotective, antiaging, anti H. pylori, neuroprotective and antinociceptive activities. In particular, the rose hip powder and extract have been reported to exert therapeutic effects on arthritis.Some of the ethnomedical indications of rose hip, such as nephroprotective and gastroproetctive actions, have been confirmed by preclinical pharmacological studies. Additional investigations on the pharmacological effects of rose hip as well as evidence from randomized controlled trials are essential to assess the therapeutic value of this natural product.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: fatty liver

Trigonelline prevents high cholesterol and high fat diet induced hepatic lipid accumulation and lipo-toxicity in C57BL/6J mice, via restoration of hepatic autophagy.

Non-alcoholic disease (NAFLD) is often linked with impaired hepatic autophagy. Here, we studied the alterations in hepatocellular autophagy by high cholesterol and high-fat diet (HC-HF) diet in C57BL/6J mice, and by (PA), in AML-12 and HepG2 cells. Further, we analysed role of Trigonelline (TG), a plant alkaloid, in preventing NAFLD, by modulating autophagy. For this, C57BL/6J mice were fed with Standard Chow (SC) or HC-HF diet, with and without TG for 16 weeks. In-vitro; AML-12\u202fcells and HepG2 cells, were exposed to PA with and without TG, for 24\u202fh. Cellular events related to autophagy, lipogenesis, and lipo-toxicity were studied. The HC-HF diet fed mice showed hepatic autophagy blockade, increased triglycerides and steatosis. PA exposure to AML-12\u202fcells and HepG2 cells induced impaired autophagy, ER stress, resulting in lipotoxicity. TG treatment in HC-HF fed mice, restored hepatic autophagy, and prevented steatosis. TG treated AML-12, and HepG2 cells exposed to PA showed autophagy restoration, and reduced lipotoxicity, however, these effects were diminished in Atg7-/- HepG2 cells, and in the presence of chloroquine. This study shows that HC-HF diet-induced impaired autophagy, and steatosis is prevented by TG, which attributes to its novel mechanism in treating NAFLD.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Nobiletin alleviates ‑induced NLRP3 inflammasome activation in a sirtuin 1‑dependent manner in AML‑12 cells.

The NOD‑like receptor family pyrin domain containing 3 (NLRP3) inflammasome has been reported to contribute to (PA)‑induced lipotoxicity. Nobiletin (Nob) is a polymethoxylated flavonoid derived from citrus fruits that has been reported to exert antioxidant and antitumor effects. However, its protective and regulatory mechanisms in PA‑induced lipotoxicity remain unclear. Therefore, the aim of the present study was to investigate the protective effects of Nob in AML‑12 cells against lipotoxicity and examine the underlying mechanism. Western blotting, reverse transcription‑quantitative polymerase chain reaction and ELISA assays were performed to investigate the activation of the NLRP3 inflammasome. Sirtuin 1 (SIRT1) small interfering RNA was used to knockdown SIRT1 expression in AML‑12 cells. The results demonstrated that PA effectively activated NLRP3 inflammasome and increased the expression and secretion of interleukin (IL)‑1β and IL‑18. Notably, the PA‑induced inflammasome activation was reversed by Nob, as indicated by the decreased expression levels of NLRP3, Caspase‑1, IL‑1β and IL‑18. Furthermore, Nob treatment with or without PA enhanced the expression of SIRT1 in AML‑12 cells, while knockdown of SIRT1 with SIRT1‑small interfering RNA reversed the anti‑inflammatory effects of Nob. Overall, the results of the present study indicated that Nob alleviated PA‑induced lipotoxicity in AML‑12 cells via the suppression of NLRP3 inflammasome activation in a SIRT1‑dependent manner. These results provide a possible basis of the underlying mechanism and, in turn, the potential application of Nob in the treatment of non‑alcoholic disease.

Keyword: fatty liver

Development of an in vitro model to study hepatitis C virus effects on hepatocellular lipotoxicity and lipid metabolism.

Hepatic steatosis is common in patients infected with hepatitis C virus (HCV). Particularly in patients infected with non-genotype 3 HCV, hepatic steatosis is closely related to factors of the metabolic syndrome such as hyperlipidemia. However, the molecular mechanisms involved in this "metabolic" steatosis in non-3 genotype HCV infections are not well understood. Here, we aimed to develop an in vitro model to study the effect of genotype 1 HCV infection on hepatic lipotoxicity and lipid metabolism. Cellular lipid accumulation was induced in Huh-7 hepatoma cells transfected with HCV genotype 1b replicon (HCV) by incubation with increasing doses of (C16:0) or oleic (C18:1 n-9) complexed to albumin mimicking hyperlipidemic conditions. Mock transfected hepatoma cells (HCV) were used as controls. Incubation with oleic concentrations as high as 0.5\u202fmM did not induce toxic effects in HCV or HCV cells. In contrast, incubation with caused dose-dependently cytotoxic effects which were more pronounced in HCV compared to HCV cells. Further analysis with subtoxic and oleic concentrations revealed a higher uptake of acids and intracellular triglyceride accumulation in HCV compared to HCV cells. Carnitine palmitoyltransferase I (CPT1) expression, indicative of mitochondrial beta-oxidation, was markedly stimulated by lipid exposure in HCV but not in HCV cells. Furthermore, heme oxygenase 1 (HMOX1) expression levels increased in FA stimulated cells, and this increase was significantly higher in HCV compared to HCV cells. In contrast, expression of the key enzymes of hepatic de novo lipogenesis synthase (FASN) and stearoyl-CoA desaturase (SCD-1) was significantly reduced upon oleate exposure in HCV but not in HCV cells. In summary, our newly developed cell culture model revealed effects of HCV genotype 1b infection on metabolic susceptibility to lipid accumulation and toxicity particularly to saturated lipids. These results may indicate that HCV (genotype 1b) infected individuals with hyperlipidemia may benefit from dietary or pharmacological intervention.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: fatty liver

Oleate ameliorates palmitate-induced reduction of NAMPT activity and NAD levels in primary human hepatocytes and hepatocarcinoma cells.

Nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide adenine dinucleotide (NAD) levels are crucial for function. The saturated palmitate and the unsaturated oleate are the main free acids in adipose tissue and human diet. We asked how these acids affect cell survival, NAMPT and NAD levels in HepG2 cells and primary human hepatocytes.HepG2 cells were stimulated with palmitate (0.5mM), oleate (1mM) or a combination of both (0.5mM/1mM) as well as nicotinamide mononucleotide (NMN) (0.5 mM) or the specific NAMPT inhibitor FK866 (10nM). Cell survival was measured by WST-1 assay and Annexin V/propidium iodide staining. NAD levels were determined by NAD/NADH Assay or HPLC. Protein and mRNA levels were analysed by Western blot analyses and qPCR, respectively. NAMPT enzyme activity was measured using radiolabelled C-nicotinamide. Lipids were stained by Oil red O staining.Palmitate significantly reduced cell survival and induced apoptosis at physiological doses. NAMPT activity and NAD levels significantly declined after 48h of palmitate. In addition, NAMPT mRNA expression was enhanced which was associated with increased NAMPT release into the supernatant, while intracellular NAMPT protein levels remained stable. Oleate alone did not influence cell viability and NAMPT activity but ameliorated the negative impact of palmitate on cell survival, NAMPT activity and NAD levels, as well as the increased NAMPT mRNA expression and secretion. NMN was able to normalize intracellular NAD levels but did not ameliorate cell viability after co-stimulation with palmitate. FK866, a specific NAMPT inhibitor did not influence lipid accumulation after oleate-treatment.Palmitate targets NAMPT activity with a consequent cellular depletion of NAD. Oleate protects from palmitate-induced apoptosis and variation of NAMPT and NAD levels. Palmitate-induced cell stress leads to an increase of NAMPT mRNA and accumulation in the supernatant. However, the proapoptotic action of palmitate seems not to be mediated by decreased NAD levels.

Keyword: fatty liver

Differential capability of metabolic substrates to promote hepatocellular lipid accumulation.

Excessive storage of triacylglycerides (TAGs) in lipid droplets within hepatocytes is a hallmark of non-alcoholic disease (NAFLD), one of the most widespread metabolic disorders in Western societies. For the purpose of exploring molecular pathways in NAFLD development and testing potential drug candidates, well-characterised experimental models of ectopic TAG storage in hepatocytes are needed.Using an optimised Oil Red O assay, immunoblotting and real-time qRT-PCR, we compared the capability of dietary monosaccharides and acids to promote lipid accumulation in HepG2 human hepatoma cells.Both high glucose and high fructose resulted in intracellular lipid accumulation after 48\xa0h, and this was further augmented (up to twofold, as compared to basal levels) by co-treatment with the lipogenesis-stimulating hormone insulin and the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-α), respectively. The acids and oleic were even more effective than these carbohydrates, inducing significantly elevated TAG storage already after 24\xa0h of treatment. Highest (about threefold) increases in lipid accumulation were observed upon treatment with oleic , alone as well as in combinations with or with high glucose and insulin. Increases in protein levels of a major lipid droplet coat protein, perilipin-2 (PLIN2), mirrored intracellular lipid accumulation following different treatment regimens.Several treatment regimens of excessive fat and sugar supply promoted lipid accumulation in HepG2 cells, albeit with differences in the extent and rapidity of steatogenesis. PLIN2 is a candidate molecular marker of sustained lipid accumulation in HepG2 cells.

Keyword: fatty liver

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model.

Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free -induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.

Keyword: fatty liver

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. histology was observed. The expression levels of genes related to lipid metabolism and proinflammation in tissue were examined. NLRP3 inflammasome expression in tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased synthesis genes, while increased oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: fatty liver

Does the enterolactone (ENL) affect transporters and lipid metabolism in ?

NAFLD as a result of inappropriate diet and obesity, may progress to sever conditions such as: type 2 diabetes mellitus or steatohepatitis, and has recently become a prevalent topic of numerous investigations. Due to its dangerous aftermaths, finding new substances, such as polyphenols and their derivatives, which might reduce steatosis is the main target of research into NAFLD treatment. Hence, the aim of the present study was to evaluate the effect(s) of enterolactone (ENL), a metabolite of secoisolariciresinol (SECO), on lipid metabolism together with changes in the expression of transporters in .The experiments were conducted on HepG2 cells incubated with either ENL and/or during 16\xa0h exposure. The expression of selected transport proteins: FATP2, FATP5, CD36, FABPpm, ABCA1, MTP, ACBP and L-FABP, as well as the proteins directly involved in lipogenesis (FAS), oxidation pathway (CPT 1), and lipid metabolism (PPARα, LXR, SREBP1c, pAMPK) was estimated by Western Blot. Intra and extracellular lipid contents were assessed by Gas-Liquid Chromatography. The data was analyzed with two-way analysis of variance (ANOVA), and results were considered to be statistically significant at \xa0≤\xa00.05.ENL stimulated extracellular efflux of free acids (FFA) and triacylglicerols (TAG) to the medium, while, it had no influence on FATP-family mediated intracellular uptake. Moreover, ENL decreased the expression of CPT 1, pAMPK, PPARα, increased SREBP1c and had no effect on LXR, and FAS content.The findings of our study demonstrate that ENL had opposite effect on steatosis in comparison with other polyphenols what suggests that it may be an inactive metabolite. ENL did not affect significantly the intracellular accumulation of FFA, DAG and TAG, yet it promoted their extracellular efflux. Furthermore, it inhibited ß-oxydation and intracellular lipid metabolism what may contribute to the progression of NAFLD.

Keyword: fatty liver

Membranotropic effects of ω-hydroxypalmitic and Ca on rat mitochondria and lecithin liposomes. Aggregation and membrane permeabilization.

The paper examines membranotropic Ca-dependent effects of ω-hydroxypalmitic (HPA), a product of ω-oxidation of acids, on the isolated rat mitochondria and artificial membrane systems (liposomes). It was established that in the presence of Ca, HPA induced aggregation of mitochondria, which was accompanied by the release of cytochrome c from the organelles. It was further demonstrated that the addition of Ca to HPA-containing liposomes induced their aggregation and/or fusion. Ca also caused the release of the fluorescent dye sulforhodamine B from liposomes, indicating their permeabilization. HPA was shown to induce a high-amplitude swelling of Ca-loaded mitochondria, to decrease their membrane potential, to induce the release of Ca from the organelles and to result in the oxidation of the mitochondrial NAD(P)H pool. Those effects of HPA were not blocked by the MPT pore inhibitor CsA, but were suppressed by the mitochondrial calcium uniporter inhibitor ruthenium red. The effects of HPA were also observed when Ca was replaced with Sr (but not with Ba or Mg). A supposition is made that HPA can induce a Ca-dependent aggregation of mitochondria, as well as Cadependent CsA-insensitive permeabilization of the inner mitochondrial membrane - with the subsequent lysis of the organelles.

Keyword: fatty liver

Ginsenoside Rg1 Ameliorates -Induced Hepatic Steatosis and Inflammation in HepG2 Cells via the AMPK/NF-B Pathway.

Nonalcoholic disease (NAFLD) is one of the common diseases in the world, and it can progress from simple lipid accumulation to sustained inflammation. The present study was designed to investigate the effects and underlying mechanisms of ginsenoside Rg1 (G-Rg1) treatment on NAFLD . HepG2 cells were treated with (PA) to induce steatosis and inflammation and then successively incubated with G-Rg1. Lipids accumulation was analyzed by Oil Red O staining and intracellular triglyceride (TG) quantification. Inflammatory conditions were examined by quantifying the levels of cell supernatant alanine transaminase/aspartate aminotransferase (ALT/AST) and secretory proinflammatory cytokines, including IL-1, IL-6, and TNF- in the cell supernatants. Quantitative RT-PCR and western blotting were used to measure the expressions of genes and proteins associated with lipogenic synthesis and inflammation, including AMP-activated protein kinase (AMPK) and nuclear factor-kappa B (NF-B) pathways. HepG2 cells were pretreated with an AMPK inhibitor; then, Oil Red O staining and TG quantification were performed to study the lipid deposition. Phospho-AMPK (Thr172) (p-AMPK) and phospho-acetyl-CoA carboxylase (Ser79) (p-ACC) were quantified by immunoblotting. Immunofluorescence was performed to demonstrate the nuclear translocation of NF-B P65. The present study showed that PA markedly increased the intracellular lipid droplets accumulation and TG levels, but decreased AMPK phosphorylation and the expressions of its downstream lipogenic genes. However, G-Rg1 alleviated hepatic steatosis and reduced the intracellular TG content; these changes were accompanied by the activation of the AMPK pathway. In addition, blocking AMPK by using the AMPK inhibitor markedly abolished the G-Rg1-mediated protection against PA-induced lipid deposition in HepG2 cells. Furthermore, G-Rg1 reduced the ALT/AST levels and proinflammatory cytokines release, which were all enhanced by PA. These effects were correlated with the inactivation of the NF-B pathway and translocation of P65 from the cytoplasm to the nucleus. Overall, these results suggest that G-Rg1 effectively ameliorates hepatic steatosis and inflammation, which might be associated with the AMPK/NF-B pathway.

Keyword: fatty liver

Dual roles of ULK1 (unc-51 like autophagy activating kinase 1) in cytoprotection against lipotoxicity.

Saturated (SFA)-induced lipotoxicity is caused by the accumulation of reactive oxygen species (ROS), which is associated with damaged mitochondria. Moreover, lipotoxicity is crucial for the progression of nonalcoholic steatohepatitis (NASH). Autophagy is required for the clearance of protein aggregates or damaged mitochondria to maintain cellular metabolic homeostasis. The NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2)-KEAP1 (kelch like ECH associated protein 1) pathway is essential for the elimination of ROS. ULK1 (unc-51 like autophagy activating kinase 1; yeast Atg1) is involved in the initiation of autophagy; however, its role in lipotoxicity-induced cell death in hepatocytes and mouse has not been elucidated. We now show that ULK1 potentiates the interaction between KEAP1 and the autophagy adaptor protein SQSTM1/p62, thereby mediating NFE2L2 activation in a manner requiring SQSTM1-dependent autophagic KEAP1 degradation. Furthermore, ULK1 is required for the autophagic removal of damaged mitochondria and to enhance binding between SQSTM1 and PINK1 (PTEN induced kinase 1). This study demonstrates the molecular mechanisms underlying the cytoprotective role of ULK1 against lipotoxicity. Thus, ULK1 could represent a potential therapeutic target for the treatment of NASH. Abbreviations: ACTB: actin beta; CM-HDCFDA:5-(and-6)-chloromethyl-2\',7\'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; LPS: lipopolysaccharides; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAPK8/JNK: mitogen-activated protein kinase 8; MEF: mouse embryonic fibroblast; MFN1: mitofusin 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NASH: nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: ; PARP: poly (ADP-ribose) polymerase 1; PINK1: PTEN induced kinase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PRKC/PKC: protein kinase C; RBX1: ring-box 1; ROS: reactive oxygen species; SFA: saturated ; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; TUBA: tubulin alpha; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase 1.

Keyword: fatty liver

Beyond the Scavenging of Reactive Oxygen Species (ROS): Direct Effect of Cerium Oxide Nanoparticles in Reducing Acids Content in an In Vitro Model of Hepatocellular Steatosis.

Nonalcoholic disease (NAFLD) is characterized by hepatic accumulation of lipids. Antisteatotic effects of cerium oxide nanoparticles (CeONPs) have recently been shown in animal models of disease. However, it is unclear whether the activity of CeONPs is related solely to the decrease in oxidative stress or, in addition, they directly decrease accumulation. To address this question, in this work, we used an in vitro model of hepatocellular steatosis, exposing HepG2 cells to oleic and . Cell uptake of CeONPs and their effect on oxidative stress and viability of hepatic cells cultured with HO were also evaluated. Results show that CeONPs were uptaken by HepG2 cells and reduced oxidative stress and improved cell viability. Treatment with oleic and increased lipogenesis and the content of different acids. CeONPs reduced and stearic and most acids consisting of more than 18 carbon atoms. These effects were associated with significant changes in elongase and desaturase activity. In conclusion, CeONPs directly protected HepG2 cells from cell injury in oxidative stress conditions and reduced content in steatotic conditions by inducing specific changes in metabolism, thus showing potential in the treatment of NAFLD.

Keyword: fatty liver

Flavone glycosides from Sicyos angulatus and their inhibitory effects on hepatic lipid accumulation.

A library of extracted natural materials (Korea Bioactive Natural Material Bank) have been screened to discover candidates for the treatment of non-alcoholic disease (NAFLD), and the 70% ethanol extract of Sicyos angulatus was found to inhibit hepatic lipid accumulation. Bioassay-guided fractionation of this bioactive extract yielded five previously undescribed flavonoid glycosides and one previously undescribed flavonolignan glycoside along with seven known flavonoid glycosides. The chemical structures of these compounds were elucidated by a combination of extensive spectroscopic analysis, including MS, NMR and UV techniques. Eight compounds of all isolated compounds showed inhibitory effects on the lipid accumulation induced by high concentrations of and glucose in HepG2 cells. Four selected compounds were tested for lipid content in a dose-dependent manner (10, 20 and 40\u202fμM), and among those compounds, kaempferol 3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside showed the strongest inhibition of hepatic lipid production in HepG2 cells. In an oil-red O staining assay, five compounds were shown to reduce hepatic lipid accumulation better than what was observed in the vehicle control group. The present study suggests a new class of chemical entities for developing bioactive agents for the treatment of diseases caused by fat accumulation in the .Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Picroside II attenuates accumulation in HepG2 cells via modulation of uptake and synthesis.

Hepatic steatosis is caused by an imbalance between free acids (FFAs) uptake, utilization, storage, and disposal. Understanding the molecular mechanisms involved in FFAs accumulation and its modulation could drive the development of potential therapies for Nonalcoholic disease. The aim of the current study was to explore the effects of picroside II, a phytoactive found in , on accumulation vis-à-vis silibinin, a known hepatoprotective phytoactive from .HepG2 cells were loaded with FFAs (oleic :/2:1) for 20 hours to mimic hepatic steatosis. The FFAs concentration achieving maximum fat accumulation and minimal cytotoxicity (500 μM) was standardized. HepG2 cells were exposed to the standardized FFAs concentration with and without picroside II pretreatment.Picroside II pretreatment inhibited FFAs-induced lipid accumulation by attenuating the expression of transport protein 5, sterol regulatory element binding protein 1 and stearoyl CoA desaturase. Preatreatment with picroside II was also found to decrease the expression of forkhead box protein O1 and phosphoenolpyruvate carboxykinase.These findings suggest that picroside II effectively attenuated accumulation by decreasing FFAs uptake and lipogenesis. Picroside II also decreased the expression of gluconeogenic genes.

Keyword: fatty liver

GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice.

Nonalcoholic disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced dysfunction, hepatic steatosis and insulin resistance. In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and lipogenesis, and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting lipogenesis and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Docosahexaenoic Reduces -Induced Endoplasmic Reticulum Stress in Pancreatic Β Cells.

Endoplasmic reticulum (ER) stress leads to peripheral insulin resistance and the progression of pancreatic beta cell failure in type 2 diabetes. Although ER stress plays an important role in the pathogenesis of diabetes, it is indispensable for cellular activity. Therefore, when assessing the pathological significance of ER stress, it is important to monitor and quantify ER stress levels. Here, we have established a novel system to monitor ER stress levels quickly and sensitively, and using this method, we have clarified the effect of differences in glucose concentration and various acids on the ER of pancreatic β cells. First, we developed a cell system that secretes Gaussia luciferase in culture medium depending on the activation of the GRP78 promoter. This system could sensitively monitor ER stress levels that could not be detected with real-time RT-PCR and immunoblotting. This system revealed that hyperglycemia does not induce unfolded protein response (UPR) in a short period of time in MIN6 cells, a mouse pancreatic β cell line. Physiological concentrations of , a saturated , induced ER stress quickly, while physiological concentrations of oleic , an unsaturated , did not. Docosahexaenoic , an n-3 unsaturated , inhibited -induced ER stress. In this study, we have established a system that can sensitively detect ER stress levels of living cells in a short period of time. This system can be used to monitor the state of the ER in living cells and lead to the investigation of the significance of physiological or pathological ER stress levels.

Keyword: fatty liver

The NOX1 isoform of NADPH oxidase is involved in dysfunction of sinusoids in nonalcoholic disease.

The increased production of reactive oxygen species (ROS) has been postulated to play a key role in the progression of nonalcoholic disease (NAFLD). However, the source of ROS and mechanisms underlying the development of NAFLD have yet to be established. We observed a significant up-regulation of a minor isoform of NADPH oxidase, NOX1, in the of nonalcoholic steatohepatitis (NASH) patients as well as of mice fed a high-fat and high-cholesterol (HFC) diet for 8 weeks. In mice deficient in Nox1 (Nox1KO), increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 demonstrated in HFC diet-fed wild-type mice (WT) were significantly attenuated. Concomitantly, increased protein nitrotyrosine adducts, a marker of peroxynitrite-induced injury detected in hepatic sinusoids of WT, were significantly suppressed in Nox1KO. The expression of NOX1 mRNA was much higher in the fractions of enriched sinusoidal endothelial cells (LSECs) than in those of hepatocytes. In primary cultured LSECs, (PA) up-regulated the mRNA level of NOX1, but not of NOX2 or NOX4. The production of nitric oxide by LSECs was significantly attenuated by PA-treatment in WT but not in Nox1KO. When the in vitro relaxation of TWNT1, a cell line that originated from hepatic stellate cells, was assessed by the gel contraction assay, the relaxation of stellate cells induced by LSECs was attenuated by PA treatment. In contrast, the relaxation effect of LSECs was preserved in cells isolated from Nox1KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impaired hepatic microcirculation through the reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the progression of NAFLD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

[Ozone oxidizes oleic with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: fatty liver

Heat shock protein 70 promotes lipogenesis in HepG2 cells.

The increasing prevalence of non-alcoholic disease (NAFLD) has followed the international rise in obesity rates. Multiple mechanisms are involved in NAFLD, including endoplasmic reticulum stress and oxidative stress. Heat shock protein 70 (HSP70), which is abundant in most organisms, is sensitive to stress. However, the role of HSP70 in NAFLD has not been investigated. Here, we investigated the possible role of HSP70 in lipid synthesis.C57BL/6 mice were fed a high-fat diet, and HepG2 cells were treated with 0.5\xa0mM (PA). HSP70 expression was detected by qPCR, Western blot and immunohistochemistry. Total cholesterol (TC) and triglyceride (TG) levels were detected by enzyme-linked immunosorbent assay (ELISA). After Hsp70 overexpression and knockdown, TC and TG levels and FAS, SCD, and ACC expression were detected.HSP70 expression was significantly increased in the livers of obese mice. In vitro, HSP70 expression was markedly induced by PA in HepG2 cells. Notably, HSP70 overexpression in HepG2 cells enhanced TC and TG synthesis, in parallel with the upregulation of lipogenic genes, including FAS, SCD and ACC. By contrast, HSP70 knockdown decreased the levels of cellular lipids and the expression of FAS, SCD, and ACC in HepG2 cells. Together, our results suggest that HSP70 may promote lipogenesis in HepG2 cells.Heat shock protein 70 promotes lipogenesis in HepG2 cells.

Keyword: fatty liver

Changes of the Profile in Erythrocyte Membranes of Patients following 6-Month Dietary Intervention Aimed at the Regression of Nonalcoholic Disease (NAFLD).

Nonalcoholic disease (NAFLD) is closely related to the metabolism disorders of acids. The pathogenesis of the disease includes an increased concentration of FFA in blood, an increase in the biosynthesis of acids, and disorders in the process of -oxidation.The aim of the study was to analyze the acids in erythrocyte membranes among 55 patients with NAFLD who were subjected to a 6-month dietary intervention in order to reduce .Basic anthropometric and biochemical measurements were performed. The profile of acids was measured in the membranes of erythrocytes and analyzed by gas chromatography. The dietary compliance was evaluated using 72-diary questionnaires, anthropometric measurements.With the reduction of (p<0.01), the patients\' biochemical and anthropometric parameters were significantly improved. A significant decrease in the concentration of alanine aminotransferase (p<0.01) and asparagine aminotransferase (p<0.01) was observed, along with a decrease in the amount of insulin (p<0.05) and insulin resistance (p<0.05). Significant changes in terms of the profile were observed among patients who followed the dietary intervention. There was a noticeable tendency in terms of the reduction (p<0.055) and a significant reduction of stearic (p<0.05). Significant changes in the profile of acids were also associated with the reductionof palmitoleic (p<0.05) and oleic acids (p<0.05). Another statistically significant change observed was the increase in polyunsaturated acids. In particular (p<0.01) the rise of eicosapentaenoic (p<0.055) and docosahexaenoic acids (p<0.55) was noted.The profile of acids turned out to be a potential biomarker of the changes during NAFLD regression. Further research is needed to fully elucidate the usefulness and applicability of our findings in the management of NAFLD.

Keyword: fatty liver

Nicotinamide riboside regulates inflammation and mitochondrial markers in AML12 hepatocytes.

The NAD precursor nicotinamide riboside (NR) is a type of vitamin B found in cow\'s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer\'s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes.A subset of hepatocytes was treated with (PA; 250 µM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 µM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed.Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes.These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.

Keyword: fatty liver

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy metabolism. However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic disease.

Keyword: fatty liver

Transport of Ca and Ca-Dependent Permeability Transition in Rat Mitochondria under the Streptozotocin-Induced Type I Diabetes.

Although diabetes mellitus is known to be a disease associated with mitochondrial dysfunction, not everything is clear about mitochondrial Ca transport and Ca-induced permeability transition in diabetic cells. The objective of this work was to study the operation of MCU and Ca-dependent mitochondrial permeabilization in the cells of Sprague-Dawley rats under the streptozotocin-induced type I diabetes. It was shown that two weeks after the induction of diabetes, the rate of Ca uptake by the mitochondria of diabetic animals increased ~1.4-fold. The expression of MCU and MICU1 subunits did not change, yet the quantity of dominant-negative MCUb channel subunits was almost twice as lower. The organelles also became more resistant to the induction of CsA-sensitive MPT pore and less resistant to the induction of CsA-insensitive palmitate/Ca-induced pore. The mitochondria of diabetic cells also showed changes in the lipid matrix of their membranes. The content of acids in the membranes grew, and microviscosity of the lipid bilayer (assessed with laurdan) increased. At the same time, lipid peroxidation (assessed by the production of malonic dialdehyde) was stimulated. The paper discusses the consequences of the diabetes-related changes in mitochondria in the context of cell physiology.

Keyword: fatty liver

[Effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes].

This study aims to explore the effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes. RAW264.7 macrophages were incubated with different kinds of acids including saturated acids- (PA), monounsaturated acids-oleic (OA) and polyunsaturated acids-docosahexaenoic (DHA), and cell culture supernatants were collected to prepare conditioned medium (CM). Hepatocytes were isolated by in situ perfusion of the with collagenase in mice, and a macrophage-hepatocyte CM co-culture system was established. Macrophage M1/M2 phenotype markers were detected by Real-time PCR. Lipid synthesis and decomposition related mRNA and protein expressions in hepatocytes were detected by Real-time PCR and Western Blot. Lipid depositions in hepatocytes were detected by oil red O staining. An analysis of variance was used for comparison of means between multiple groups. Compared with control groups, PA polarized macrophages to a M1 phenotype (expression of TNF-α and IL-6 significantly increased, F≥22.68, P < 0.01), OA polarized macrophages to a M1/M2 mixed phenotype (expression of IL-6, Mrc2 and IL-10 increased F≥4.94, P < 0.05) and DHA polarized macrophages to a M2 phenotype (expression of Mrc2 and IL-10 significantly increased, F≥4.94, P < 0.01). CM-PA significantly increased lipid synthesis related genes, including SREBP1C, ACC1 mRNA expression (F≥5.66, P < 0.01) and FASN, ACC1 protein expression (F≥38.34, P < 0.05) in hepatocytes, and decreased lipid decomposition gene ACOX1 protein expression (F=154.48, P < 0.01). CM-OA affected several lipid metabolism genes expression. CM-DHA significantly increased CPT1A mRNA expression (F = 10.30, P < 0.01) and ACOX1, CPT1A protein expression (F≥47.06, P < 0.05), and decreased SREBP1C, ACC1 protein expression (F≥65.84, P < 0.05) in hepatocytes. Massive lipid droplets were deposited in hepatocytes in CM-PA treated hepatocytes, and a few amount of lipid droplets were deposited in CM-DHA treated hepatocytes. Different acids affect the balance of lipid metabolism in hepatocytes and by inducing macrophage M1 / M2 polarization, thus promoting or delaying the progression of non-alcoholic disease.

Keyword: fatty liver

Upregulation of miR-181a impairs lipid metabolism by targeting PPARα expression in nonalcoholic disease.

Recent studies have reported elevated expression of miR-181a in patients with non-alcoholic disease (NAFLD), suggesting that it may play an important role in lipid metabolism and insulin resistance. We aimed to investigate the effect of miR-181a in lipid metabolism and find new treatments for NAFLD. The expression level of miR-181a in NAFLD patient serum and a (PA)-induced NAFLD cell model was examined by Q-PCR. Oil red O staining and triglyceride assays were used to assess lipid accumulation in hepatocytes. Western blotting was used to detect the protein expression levels of peroxisome proliferator-activated receptor-α (PPARα) and the β-oxidation-related genes. Direct interactions were validated by dual-luciferase reporter gene assays. MiR-181a expression was significantly upregulated in the serum of NAFLD patients and PA-induced hepatocytes. Inhibition of miR-181a expression resulted in the increased expression of PPARα and its downstream genes, and PA-induced lipid accumulation in hepatocytes was also inhibited. Upregulation of miR-181a resulted in the downregulation of its direct target PPARα and downstream gene expression of PPARα as well as aggravated lipid accumulation in hepatocytes. At the same time, the increased expression of PPARα can offset lipid accumulation in hepatocytes induced by miR-181a mimics. This study demonstrates that reducing the expression of miR-181a may improve lipid metabolism in NAFLD. The downregulation of miR-181a expression can be a therapeutic strategy for NAFLD by modulating its target PPARα.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Effects of saturated and omega-3 polyunsaturated acids on Sertoli cell apoptosis.

Obesity is believed to negatively affect male semen quality and is accompanied by dysregulation of free (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report obesity decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive oxygen species were elevated. Furthermore, we demonstrated by in vitro assays that saturated (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated acids might be the cause of obesity-induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs.FFA: free ; HFD: high-fat diet; SD: standard diet; PA: ; PUFA: polyunsaturated ; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive oxygen species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic disease; DCFH-DA: 2\', 7\' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic ; PI: propidium iodide; DHA: docosahexenoic .

Keyword: fatty liver

activates NLRP3 inflammasomes in mouse Kupffer cells through mitochondrial DNA release.

Nonalcoholic disease (NAFLD) is the most common disease in many developed and developing countries worldwide. It has been well established that the chronic sterile inflammation caused by the NLRP3 inflammasome is closely related to NAFLD development. Kupffer cells (KCs) are involved in the pathogenesis of various diseases. We used methionine choline-deficient diets to establish a mouse nonalcoholic steatohepatitis (NASH) model. The expression and formation of the NLRP3 inflammasome in the KCs from the mouse and cell models were determined by Western blotting and co-immunoprecipitation. Evidence of mitochondrial DNA (mtDNA) release was determined by live cell labeling and imaging. KCs and the NLRP3 inflammasome exerted proinflammatory effects on the development and progression of NASH through secretion of the proinflammatory cytokine IL-1β. NLRP3, ASC and Caspase-1 protein expression levels in KCs from NASH mouse livers were significantly higher than those in KCs from NLRP3 mice, and the number of NLRP3 inflammasome protein complexes was significantly higher in KCs from NASH mouse livers, whereas these protein complexes could not be formed in NLRP3 mice. In in vitro experiments, (PA) decreased the mitochondrial membrane potential and subsequently induced mtDNA release from the mitochondria to the cytoplasm. NLRP3 inflammasome expression was substantially increased, and mtDNA-NLRP3 inflammasome complexes formed upon PA stimulation. Our data suggest that mtDNA released from mitochondria during PA stimulation causes NLRP3 inflammasome activation, providing a missing link between NLRP3 inflammasome activation and NASH development, via binding of cytosolic mtDNA to the NLRP3 inflammasome.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis.

Mitochondrial dysfunction plays a crucial role in the development of non-alcoholic steatohepatitis (NASH). However, the regulator of mitochondrial dysfunction in the pathogenesis of NASH is still largely unclear. CXCR3 is an essential pro-inflammatory factor in chronic diseases. We explored the significance of CXCR3 in regulating mitochondrial function during NASH development in animal models and cultured hepatocytes.The effects of CXCR3 on mitochondrial function were evaluated by genetic knockout or pharmacological inhibition in mouse models and . The ultrastructural changes of mitochondria were assessed by transmission electron microscopy (TEM). Hepatic levels of mitochondrial reactive oxygen species (ROS), DNA damage, membrane potential and ATP were examined.CXCR3 ablation by genetic knockout or pharmacological inhibition in mice protected against NASH development by influencing mitochondrial function. Similarly, depletion of CXCR3 reduced steatohepatitis injury in cultured hepatocytes. TEM analysis revealed that mitochondrial integrity was much improved in CXCR3 knockout (CXCR3) compared to wildtype (WT) mice. In agreement with this, impaired mitochondrial function was pronounced in WT mice compared to CXCR3 mice, evidenced by increased protein expression of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and decreased protein expression of mitofusin-1 (MFN1). Mitochondrial dysfunction was induced in AML-12 hepatocytes by methionine and choline deficient medium and in HepG2 cells by . The impaired mitochondrial function in both cell lines was evidenced by reduced membrane potential and ATP content, and by increased mitochondrial ROS accumulation and DNA damage. However, CXCR3 knockdown by siCXCR3 significantly diminished the mitochondrial dysfunction in both AML-12 and HepG2 hepatocytes. In addition, inhibition of CXCR3 by CXCR3 specific antagonists SCH546738 and AMG487 restored mitochondrial function and inhibited mitochondrial-dependent apoptosis in the of WT mice fed with methionine and choline deficient diet.CXCR3 induces mitochondrial dysfunction, which contributes to the pathogenesis of steatohepatitis. Pharmacologic blockade of CXCR3 prevents mitochondrial dysfunction and restores the severity of steatohepatitis, indicating a potential clinical impact for controlling the disease.

Keyword: fatty liver

Mst1 inhibition attenuates non-alcoholic disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high-fat diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fatty liver

γδ T cells Promote Steatohepatitis by Orchestrating Innate and Adaptive Immune Programming.

The recruitment and activation of inflammatory cells in the delineates the transition from hepatic steatosis to steatohepatitis. We found that in steatohepatitis, γδT cells are recruited to the by CCR2, CCR5, and NOD2 signaling and are skewed towards an IL-17A phenotype in an ICOS-ICOSL dependent manner. γδT cells exhibit a distinct Vγ4 , PD1 , Ly6C CD44 phenotype in steatohepatitis. Moreover, γδT cells upregulate both CD1d, which is necessary for lipid-based antigens presentation, and the free receptor CD36. γδT cells are stimulated to express IL-17A by and CD1d ligation. Deletion, depletion, and targeted interruption of γδT cell recruitment protects against diet-induced steatohepatitis and accelerates disease resolution. We demonstrate that hepatic γδT cells exacerbate steatohepatitis, independent of IL-17 expression, by mitigating conventional CD4 T cell expansion and modulating their inflammatory program via CD1d-dependent VEGF expression.© 2019 by the American Association for the Study of Diseases.

Keyword: fatty liver

Hsp72 protects against injury via attenuation of hepatocellular death, oxidative stress, and JNK signaling.

Heat shock protein (Hsp) 72 is a molecular chaperone that has broad cytoprotective functions and is upregulated in response to stress. To determine its hepatic functions, we studied its expression in human disorders and its biological significance in newly generated transgenic animals.Double transgenic mice overexpressing Hsp72 (gene Hspa1a) under the control of a tissue-specific tetracycline-inducible system (Hsp72-LAP mice) were produced. Acute injury was induced by a single injection of acetaminophen (APAP). Feeding with either a methionine choline-deficient (MCD; 8\u202fweeks) or a 3,5-diethoxycarbonyl-1,4-dihydrocollidine-supplemented diet (DDC; 12\u202fweeks) was used to induce lipotoxic injury and Mallory-Denk body (MDB) formation, respectively. Primary hepatocytes were treated with .Patients with non-alcoholic steatohepatitis and chronic hepatitis C infection displayed elevated HSP72 levels. These levels increased with the extent of hepatic inflammation and HSP72 expression was induced after treatment with either interleukin (IL)-1β or IL-6. Hsp72-LAP mice exhibited robust, hepatocyte-specific Hsp72 overexpression. Primary hepatocytes from these animals were more resistant to isolation-induced stress and Hsp72-LAP mice displayed lower levels of hepatic injury in vivo. Mice overexpressing Hsp72 had fewer APAP protein adducts and were protected from oxidative stress and APAP-/MCD-induced cell death. Hsp72-LAP mice and/or hepatocytes displayed significantly attenuated Jnk activation. Overexpression of Hsp72 did not affect steatosis or the extent of MDB formation.Our results demonstrate that HSP72 induction occurs in human disease, thus, HSP72 represents an attractive therapeutic target owing to its broad hepatoprotective functions.HSP72 constitutes a stress-inducible, protective protein. Our data demonstrate that it is upregulated in patients with chronic hepatitis C and non-alcoholic steatohepatitis. Moreover, Hsp72-overexpressing mice are protected from various forms of stress.Copyright © 2018 European Association for the Study of the . All rights reserved.

Keyword: fatty liver

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: fatty liver

Omega-3 polyunsaturated acids alleviate hepatic steatosis-induced inflammation through Sirt1-mediated nuclear translocation of NF-κB p65 subunit in hepatocytes of large yellow croaker (Larmichthys crocea).

Hepatic steatosis induced inflammation is becoming increasingly prevalent in farmed fish. This study was conducted to investigate the protective effects of omega-3 polyunsaturated acids (ω-3 PUFAs) against hepatic steatosis-induced inflammation and its potential molecular mechanisms in hepatocyte of large yellow croaker (Larmichthys crocea). We found that the hepatic steatosis-induced inflammation was relieved by ω-3 PUFAs, meanwhile, the Sirt1 activity and transcript expression was increased by ω-3 PUFAs. The increased Sirt1 activity can decrease the hepatic steatosis-induced inflammation. The protective effects of ω-3 PUFAs against hepatic steatosis-induced inflammation was reversed by the treatment with Sirt1 inhibitor EX-527. The nuclear translocation of nuclear transcription factor kappa-B (NF-κB) p65 was significantly decreased after ω-3 PUFAs treatments compared to the stimulation group. The ω-3 PUFAs induced cytoplasm translocation of NF-κB p65 was reversed by EX-527. Together, ω-3 PUFAs alleviate hepatic steatosis-induced inflammation through Sirt1-mediated nuclear translocation of NF-κB p65 subunit in hepatocytes of large yellow croaker. The present study provides important insight into the mechanisms of the protective effects of ω-3 PUFAs, providing theory bases for alleviating the hepatic steatosis induced inflammation of farmed fish, thereby offering great benefits to the aquaculture industry and fish consumers.Copyright © 2017. Published by Elsevier Ltd.

Keyword: fatty liver

Long-chain activates hepatocytes through CD36 mediated oxidative stress.

Accumulating evidence suggests that activated hepatocytes are involved in the deposition of the excess extracellular matrix during fibrosis via the epithelial to mesenchymal transition. Lipid accumulation in hepatocytes are implicated in the pathogenesis of chronic injury. CD36 is known to mediate long-chain (LCFA) uptake and lipid metabolism. However, it is unclear whether LCFA directly promotes hepatocyte activation and the involved mechanisms have not been fully clarified.Mice were fed with a high fat diet (HFD) and normal hepatocyte cells (Chang cells) were treated with (PA) in vivo and in vitro. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to examine the gene and protein expression of molecules involved in hepatic fibrogenesis and hepatocyte activation. CD36 was knocked down by transfecting CD36 siRNA into hepatocyte cells. Hydrogen peroxide (HO) and reactive oxygen species (ROS) levels were detected using commercial kits.HFD induced a profibrogenic response and up-regulated CD36 expression in vivo. Analogously, PA increased lipid accumulation and induced human hepatocyte activation in vitro, which was also accompanied by increased CD36 expression. Interestingly, knockdown of CD36 resulted in a reduction of hepatocyte lipid deposition and decreased expression of Acta2 (34% decrease), Vimentin (29% decrease), Desmin (60% decrease), and TGF-β signaling pathway related genes. In addition, HFD and PA increased the production of HO in vivo (48% increase) and in vitro (385% increase), and the antioxidant, NAC, ameliorated PA-induced hepatocyte activation. Furthermore, silencing of CD36 in vitro markedly attenuated PA-induced oxidative stress (HO: 41% decrease; ROS: 39% decrease), and the anti-activation effects of CD36 knockdown could be abolished by pretreatment with HO.Our study demonstrated that LCFA facilitates hepatocyte activation by up-regulating oxidative stress through CD36, which could be an important mechanism in the development of hepatic fibrosis.

Keyword: fatty liver

Adaptations of hepatic lipid metabolism and mitochondria in dairy cows with mild .

The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and mitochondria in dairy cows with mild , and blood samples were collected from healthy cows (n = 15) and cows with mild (n = 15). To determine the effects of acids (PA), one of the major component of acids, on lipid metabolism and mitochondria in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM). Dairy cows with mild displayed hepatic lipid accumulation. The protein levels of sterol regulatory element-binding protein 1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPARα) and mRNA levels of acetyl CoA carboxylase 1 (ACC1), synthase (FAS), acyl-CoA oxidase (ACO), and carnitine palmitoyltransferase 1A (CPT1A) were significantly higher in dairy cows with mild than in control cows. The hepatic mitochondrial DNA content, mRNA levels of oxidative phosphorylation complexes I to V (CO 1-V), protein levels of cytochrome c oxidase subunit IV (COX IV), voltage dependent anion channel 1 (VDAC1), peroxisome proliferator activated receptor-γ coactivator-1α (PGC-1α) and nuclear respiratory factor 1 (NRF1), and adenosine triphosphate (ATP) content were all markedly increased in the of dairy cows with mild compared with healthy cows. The PA treatment significantly increased lipid accumulation; protein levels of SREBP-1c and PPARα; and mRNA levels of ACC1, FAS, ACO, and CPT1A in calf hepatocytes. Moreover, the mitochondrial DNA content, mRNA levels of CO 1-V, protein levels of COX IV, VDAC1, PGC-1α, NRF1, mitochondrial transcription factor A, and ATP content were significantly increased in PA-treated hepatocytes compared with control hepatocytes. The protein level of mitofusin-2 was significantly decreased in PA-treated groups. In conclusion, lipid synthesis and oxidation, number of mitochondria, and ATP production were increased in the of dairy cows with mild and PA-treated calf hepatocytes. These changes in hepatic mitochondria and lipid metabolism may be the adaptive mechanism of dairy cows with mild .Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

MicroRNA-194 inhibition improves dietary-induced non-alcoholic disease in mice through targeting on FXR.

Non-alcoholic disease (NAFLD) affects obesity-associated metabolic syndrome, which exhibits hepatic steatosis, insulin insensitivity and glucose intolerance. Previous studies indicated that hepatic microRNAs (miRs) play critical roles in the development of NAFLD. In this study, we aim to explore the pathophysiological role of miR-194 in obesity-mediated metabolic dysfunction. Our findings show that the high fat diet or treatment significantly increase hepatic miR-194 levels in vivo and in vitro. Silence of miR-194 protects -induced inflammatory response in cultured hepatocytes, and attenuates structural disorders, lipid deposits and inflammatory response in . MiR-194 inhibitor also improves glucose and insulin intolerance in obese mice. Through dual luciferase assay, we demonstrate that miR-194 directly binds to FXR/Nr1h4 3\'-UTR, and inhibits gene expression of FXR/Nr1h4. Furthermore, overexpression of miR-194 downregulates FXR/Nr1h4 in cultured hepatocytes, but miR-194 inhibitor reversely increases FXR/Nr1h4 expression in obese mouse tissues. On the contrast, silence of FXR/Nr1h4 abolishes the hepatic benefits in obese mice treated with miR-194 inhibitor. Present study provides a novel finding that suppression of miR-194 attenuates dietary-induced NAFLD via upregulation of FXR/Nr1h4. The findings suggest miR-194/FXR are potential diagnostic markers and therapeutic targets for NAFLD.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Resveratrol Ameliorates Lipid Droplet Accumulation in Through a SIRT1/ ATF6-Dependent Mechanism.

Lipid droplets (LDs) are dynamic organelles that store neutral lipids during times of energy excess, and an increased accumulation of LDs in the is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high-fat diet (HFD) and were used to induce hepatic steatosis in mouse and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Insulin-like growth factor binding protein 7 accelerates hepatic steatosis and insulin resistance in non-alcoholic fatty liver disease.

An association between increased insulin-like growth factor binding protein-7 (IGFBP7) expression and insulin resistance in metabolic diseases has been reported. However, the role and molecular mechanism of IGFBP-7 in non-alcoholic fatty liver disease (NAFLD) remains largely unknown. Therefore, the potential function of IGFBP7 in the pathological progression of NAFLD was explored in this investigation. For in vivo experiments, an animal model of NAFLD was established in C57BL/6 mice by feeding a high-fat diet (HFD), and IGFBP7 was knocked down by injecting adeno-associated adenovirus (AAV)-mediated short-hairpin (sh)-IGFBP7 into the liver. We found that AAV-sh-IGFBP7 treatment significantly alleviated hepatocyte injury and inhibited hepatic lipid accumulation by reducing lipogenesis-associated gene expression. Furthermore, downregulation of IGFBP7 markedly ameliorated IR and restored impaired insulin signalling by elevating the phosphorylation levels of IRS-1, Akt and GSK3β in HFD-treated mice. Similar results were also confirmed by an in vitro study in a (PA)-stimulated HepG2 cell model. In conclusion, our study demonstrates that IGFBP7 contributes to hepatic steatosis and insulin resistance in NAFLD development, which might serve as a novel therapeutic agent for the treatment of NAFLD.© 2019 John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

Systematic evaluation of phenolic compounds and protective capacity of a new mulberry cultivar J33 against -induced lipotoxicity using a simulated digestion method.

This research aimed to investigate the protective effects of a new mulberry cultivar J33 with simulated gastrointestinal digestion against (PA)-induced lipotoxicity. LC-MS analysis revealed that the contents of four flavonoid glycosides (quercetin rhamnosylhexoside hexoside, quercetin rhamnosylhexoside, quercetin hexoside, kaempferol rhamnosylhexoside) increased after digestion. Besides, mulberry digest (MBD) at 0.5-2\u202fmg/mL significantly reduced PA-induced lipotoxicity in human hepatocytes, while mulberry extract without digestion (MBE) showed no protection. Further investigations demonstrated that the protection of MBD was attributed to two aspects. On the one hand, MBD could attenuate PA-induced oxidative stress by suppressing ROS accumulation, regulating intracellular glutathione and ameliorating mitochondrial dysfunction. On the other hand, MBD could promote PA incorporation into inert triglycerides (TG) to deal with the acute lipid overload, reducing the lipotoxicity caused by PA. Overall, our research might provide a new perspective of mulberry cultivar J33 in ameliorating non-alcoholic disease (NAFLD).Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

PKCδ silencing alleviates saturated induced ER stress by enhancing SERCA activity.

Protein kinase C δ (PKCδ) plays an important role in nonalcoholic disease (NAFLD), however, the mechanism remains unknown. The present study explored the role of PKCδ in NAFLD development and investigated the relationships between PKCδ, calcium homeostasis, and endoplasmic reticulum (ER) stress (ERS). Hepatic steatosis cell model was induced by (PA) in L02 cells. Lipid accretion was evaluated using Oil Red O staining and a triglyceride (TG) detection kit. PKCδ was down-regulated by siRNA. RT-PCR and Western blotting were used to detect the expression of ERS markers. The fluorescence of Ca influx was recorded using confocal microscopy. Sarco-ER Ca-ATPase (SERCA) activity was measured by ultramicro-ATP enzyme test kit. PA treatment induced lipid accretion in L02 cells, destroyed the ER structure, and increased PKCδ activation in a time-dependent manner. Further, PA treatment significantly increased the expression of ERS markers, Ig heavy chain binding protein (Bip), and homologous proteins of CCAAT-enhancer binding proteins (CHOP). PKCδ silencing down-regulated Bip and CHOP expression, indicating a successful alleviation of ERS. The increased calcium storage induced by PA stimulation was significantly decreased in L02 cells treated with PKCδ siRNA compared with the negative control. Moreover, diminished SERCA activity caused by PA was recovered in PKCδ siRNA transfected cells. To the best of our knowledge, this is the first report demonstrating that the inhibition of PKCδ alleviates ERS by enhancing SERCA activity and stabilizing calcium homeostasis.© 2017 The Author(s).

Keyword: fatty liver

The Hepatotoxicity of in Zebrafish Involves the Intestinal Microbiota.

(PA) is the main saturated naturally occurring in animal fats and vegetable oils. In recent decades, palm oil, an alternative lipid source containing high amounts of PA, has been widely used to replace fish oil in aquafeed.We investigated the hepatotoxicity of PA in zebrafish and the underlying mechanism.One-month-old zebrafish fed a high-fat diet (HFD) containing 16% soybean oil and 3 PA-incorporated HFDs [4%, 8%, and 12% PA (12PA)] for 2 wk (experiment 1) and 4 wk (experiment 2) were used to evaluate PA-induced damage and endoplasmic reticulum (ER) stress. Germ-free (GF) zebrafish fed low-fat, high-fat, or 12PA diets for 5 d were used to study the direct effects of PA on damage (experiment 3). GF zebrafish colonized with HFD or 12PA microbiota for 48 h were used to elucidate the indirect effects of PA-altered microbiota on damage (experiment 4). Last, GF zebrafish colonized with HFD or 12PA microbiota were used to evaluate the effects of different microbiotas on PA absorption (experiment 5).In experiment 1, the proportion of PA in the linearly increased as its percentage in dietary lipid increased (r2\xa0=\xa00.83, P\xa0<\xa00.05). In experiment 2, the expression of glucose-regulated protein 78 (Grp78) and C/EBP-homologous protein (Chop) was higher in the 12PA group than in the HFD group (2.2- and 2.7-fold, respectively; P\xa0<\xa00.05). The activity of caspase-12 was increased by 61.1% in the 12PA group compared with the HFD group (P\xa0<\xa00.05). In experiment 3, caspase-12 activity was higher in the 12PA group than in the HFD group (P\xa0<\xa00.05). In experiment 4, GF zebrafish colonized with PA-altered microbiota had higher caspase-12 activity (P\xa0<\xa00.05) than those colonized by HFD microbiota. In experiment 5, PA-altered microbiota promoted PA absorption (P\xa0<\xa00.05) and aggravated ER stress and damage in the context of high-PA feeding.The PA-altered microbiota indirectly induced ER stress and damage in zebrafish. Moreover, the PA microbiota promoted the absorption of PA, leading to enhanced PA overflow into the and aggravated hepatotoxicity of PA in zebrafish.

Keyword: fatty liver

Analysis of proautophagic activities of Citrus flavonoids in cells reveals the superiority of a natural polyphenol mixture over pure flavones.

Autophagy dysfunction has been implicated in the pathogenesis of nonalcoholic disease (NAFLD). Natural compounds present in bergamot polyphenol fraction (BPF) prevent NAFLD and induce autophagy in rat livers. Here, we employed HepG2 cells expressing DsRed-LC3-GFP, a highly sensitive model system to screen for proautophagic compounds present in BPF. BPF induced autophagy in a time- and dose-dependent fashion and the effect was amplified in cells loaded with . Autophagy was mediated by the hydrophobic fraction of -hydrolyzed BPF (A-BPF), containing six flavanone and flavone aglycones as identified by liquid chromatography-high-resolution mass spectrometry. Among them, naringenin, hesperitin, eriodictyol and diosmetin were weak inducers of autophagy. Apigenin showed the strongest and dose-dependent proautophagic activity at early time points (6 h). Luteolin induced a biphasic autophagic response, strong at low doses and inhibitory at higher doses. Both flavones were toxic in HepG2 cells and in differentiated human progenitors HepaRG upon longer treatments (24 h). In contrast, BPF and A-BPF did not show any toxicity, but induced a persistent increase in autophagic flux. A mixture of six synthetic aglycones mimicking A-BPF was sufficient to induce a similar autophagic response, but it was mildly cytotoxic. Thus, while six main BPF flavonoids fully account for its proautophagic activity, their combined effect is not sufficient to abrogate cytotoxicity of individual compounds. This suggests that a natural polyphenol phytocomplex, such as BPF, is a safer and more effective strategy for the treatment of NAFLD than the use of pure flavonoids.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

The beneficial effects of resveratrol on steatosis and mitochondrial oxidative stress in HepG2 cells.

Nonalcoholic disease (NAFLD) is currently one of the most common chronic diseases, especially in developed countries. One group of substances with a potential use in the treatment of NAFLD are plant polyphenols, represented by resveratrol. The aim of this study was to evaluate the effect of resveratrol on steatosis and oxidative stress in HepG2 cells. The steatosis of cells was carried out using free acids: oleic or and their mixtures. Steatosis was visualized using the intracellular lipid staining by Nile Red dye with a fluorescence microscope. This study also determined the viability of cells and mitochondrial membrane potential. The current study showed that acids and their mixtures induced fat overloading in HepG2 cells. In the group of cells incubated with oleic (OA), observed changes were moderate with prevailing micro-vesicular steatosis. In case of cells incubated with (PA) and the mixtures of acids, micro- and macro-vacuolar steatosis occurred in most of the cells. Resveratrol decreased steatosis in HepG2 cells induced by OA, PA, as well as their mixtures, and in most of experimental groups did not reduce cells viability. Resveratrol reduced the oxidative stress in HepG2 cells treated with acids mixtures.

Keyword: fatty liver

Liraglutide ameliorates non-alcoholic steatohepatitis by inhibiting NLRP3 inflammasome and pyroptosis activation via mitophagy.

Non-alcoholic steatohepatitis (NASH) is a key step in the progression of non-alcoholic disease (NAFLD), which causes serious health problems worldwide. The nucleotide-binding oligomerization domain, leucine-rich repeat-containing receptor-containing pyrin domain 3 (NLRP3) inflammasome and pyroptosis play crucial roles in the progression of NASH. Our team has provided clinical evidence of the effects of glucagon-like peptide-1 (GLP-1) on the improvement in function and histological resolution of NAFLD. Preliminary work has demonstrated that GLP-1 inhibited NLRP3 inflammasome activation in a mouse model of NAFLD. We further explored the potential molecular mechanisms underlying the anti-inflammatory effect of liraglutide, a long-acting GLP-1 analog, in the treatment of NASH. We established a HepG2 cell model of NASH using double stimulation with and lipopolysaccharide to assess NLRP3 inflammasome and pyroptotic cell activity and to evaluate mitochondrial function and mitophagy. Liraglutide reduced lipid accumulation, inhibited NLRP3 inflammasome and pyroptosis activation, attenuated mitochondrial dysfunction and reactive oxygen species generation, augmented mitophagy in hepatocytes. Mitophagy inhibition with 3-methyladenine/PINK1-directed siRNA weakened the liraglutide-mediated suppression of inflammatory injury. We propose that liraglutide suppresses NLRP3 inflammasome-induced hepatocyte pyroptosis via mitophagy to slow the progression of NASH.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Inhibition of hepatocyte nuclear factor 1b induces hepatic steatosis through DPP4/NOX1-mediated regulation of superoxide.

Nonalcoholic disease (NAFLD) is the most common chronic disorder that is closely associated with insulin resistance and type 2 diabetes. Previous studies have suggested that hepatocyte nuclear factor 1b (HNF1b) ameliorates insulin resistance. However, the role of HNF1b in the regulation of lipid metabolism and hepatic steatosis remains poorly understood. We found that HNF1b expression was decreased in steatotic livers. We injected mice with lentivirus (LV) expressing HNF1b shRNA to generate mice with hepatic knockdown of HNF1b. We also injected high fat (HF) diet-induced obese and db/db diabetic mice with LV expressing HNF1b to overexpress HNF1b. Knockdown of HNF1b increased hepatic lipid contents and induced insulin resistance in mice and in hepatocytes. Knockdown of HNF1b worsened HF diet-induced increases in hepatic lipid contents, injury and insulin resistance in mice and PA-induced lipid accumulation and impaired insulin signaling in hepatocytes. Moreover, overexpression of HNF1b alleviated HF diet-induced increases in hepatic lipid content and insulin resistance in mice. Knockdown of HNF1b increased expression of genes associated with lipogenensis and endoplasmic reticulum (ER) stress. DPP4 and NOX1 expression was increased by knockdown of HNF1b and HNF1b directly bound with the promoters of DPP4 and NOX1. Overexpression of DPP4 or NOX1 was associated with an increase in lipid droplets in hepatocytes and decreased expression of DPP4 or NOX1 suppressed the effects of knockdown of HNF1b knockdown on triglyceride (TG) formation and insulin signaling. Knockdown of HNF1b increased superoxide level and decreased glutathione content, which was inhibited by downregulation of DPP4 and NOX1. N-acetylcysteine (NAC) suppressed HNF1b knockdown-induced ER stress, TG formation and insulin resistance. (PA) decreased HNF1b expression which was inhibited by NAC. Taken together, these studies demonstrate that HNF1b plays an essential role in controlling hepatic TG homeostasis and insulin sensitivity by regulating DPP4/NOX1mediated generation of superoxide.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Cangju Qinggan Jiangzhi Decoction Reduces the Development of NonAlcoholic Steatohepatitis and Activation of Kupffer Cells.

Nonalcoholic steatohepatitis (NASH) is defined as lipid accumulation with hepatic injury, inflammation and early to moderate fibrosis. Kupffer cells play a crucial role in promoting hepatic inflammation, which further facilitates the development of NASH. Here we investigated the effects of Cangju Qinggan Jiangzhi decoction (CQJD) on high fat diet (HFD) and methionine-choline deficient (MCD) induced mouse NASH pathogenesis.Mouse NASH models were developed by HFD and MCD diet. The treated mice were divided into three groups: the control group (n = 10), the low-dose CQJD treatment group (n = 10) and the high-dose CQJD treatment group (n = 10). The hepatic injury, inflammation, and apoptotic molecules were evaluated by H&E staining, immunohistochemistry and real-time PCR. Kupffer cells were isolated from control mice and CQJD-treated mice after stimulation by lipopolysaccharide (LPS) and/or . The level of the inflammatory cytokines TNFα, IL1β, and CCL2 was measured by ELISA.The HFD-fed mice displayed significant metabolic, inflammatory, and oxidative stress-related alterations due to hepatic lipid accumulation. CQJD treatment largely normalized the hepatic injury, lowered the ALT/AST level, and reduced the severity of inflammation, as revealed by the decreased inflammatory cytokines levels. In vitro, CQJD blocked the activation of LPS- or -primed Kupffer cells in a dose-dependent manner. In the MCD diet-induced NASH mice, similar therapeutic effects of CQJD were also observed.CQJD ameliorates mouse nonalcoholic steatohepatitis. The reduction in injury and inflammation induced by CQJD is associated with reduced activation of Kupffer cells. Our results suggest that CQJD is a promising therapeutic strategy in clinical steatohepatitis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Effects of abomasal infusions of acids and one-carbon donors on hepatic ceramide and phosphatidylcholine in lactating Holstein dairy cows.

Our objectives were to (1) determine whether the abomasal infusion of behenic (C22:0) elevated hepatic ceramide relative to (C16:0) or docosahexaenoic (C22:6n-3) infusion; (2) assess whether the abomasal infusion of choline chloride or l-serine elevated hepatic phosphatidylcholine (PC) in cows abomasally infused with C16:0; and (3) characterize the PC lipidome in cows abomasally infused with C22:6n-3, relative to C16:0 or C22:0 infusion. In a 5 × 5 Latin square design, 5 rumen-cannulated Holstein cows (214 ± 4.9 DIM; 3.2 ± 1.1 parity) were enrolled in a study with 6-d periods. Abomasal infusates consisted of (1) (PA; 98% C16:0); (2) PA + choline chloride (PA+C; 50 g/d choline chloride); (3) PA + l-serine (PA+S; 170 g/d l-serine); (4) behenic (BA; 92% C22:0); and (5) an algal oil rich in docosahexaenoic (DHA; 44% C22:6n-3). Emulsion infusates provided 301 g/d of total acids containing a minimum of 40 g/d of C16:0. Cows were fed a corn silage-based diet. Milk was collected on d -2, -1, 5, and 6. Blood was collected and biopsied on d 6 of each period. Although we did not detect differences in milk yield, milk fat yield and content were lower in cows infused with DHA relative to PA. Plasma triacylglycerol concentrations were lower with DHA treatment relative to PA or BA. Cows infused with DHA had lower plasma insulin concentrations relative to cows infused with PA only. For objective 1, hepatic ceramide-d18:2/22:0 was highest in cows infused with BA relative to other treatments. For objective 2, plasma free choline concentrations were greater in PA+C cows relative to PA; however, we did not observe this effect with PA+S. Plasma total PC concentrations were similar for all treatments. Regarding the hepatic lipidome, a total of 18 hepatic PC were higher (e.g., PC-16:1/18:2) and 25 PC were lower (e.g., PC-16:0/22:6) with PA+C infusion relative to PA. In addition, 17 PC were higher (e.g., PC-20:3/22:5) and 21 PC were lower (e.g., PC-18:0/22:6) with PA+S infusion relative to PA. For objective 3, hepatic concentrations of many individual saturated PC (e.g., PC-18:0/15:0) were lower with DHA relative to other treatments. Hepatic concentrations of highly unsaturated PC with very-long-chain acids (e.g., PC-14:0/22:6) were higher in DHA-infused cows relative to PA, PA+C, PA+S, or BA. The abomasal infusion of emulsions containing , with choline chloride or serine, behenic , or docosahexaenoic influence the hepatic ceramide and PC profiles of lactating cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

Celastrol reverses (PA)-caused TLR4-MD2 activation-dependent insulin resistance via disrupting MD2-related cellular binding to PA.

Elevated plasma statured acids (FFAs) cause TLR4/MD2 activation-dependent inflammation and insulin tolerance, which account for the occurrence and development of obesity. It has been confirmed that statured (PA) (the most abundant FFA) could bind MD2 to cause cellular inflammation. The natural compound celastrol could improve obesity, which is suggested via inhibiting inflammation, yet the detailed mechanism for celastrol is still unclear. As celastrol is reported to directly target MD2, we thought disrupting the binding between FFAs and MD2 might be one of the ways for celastrol to inhibit FFAs-caused inflammation and insulin resistance. In this study, we found evidence to support our hypothesis: celastrol could reverse PA-caused TLR4/MD2 activation-dependent insulin resistance, as determined by glucose-lowering ability, cellular glucose uptake, insulin action-related proteins and TLR4/MD2/NF-κB activation. Bioinformatics and cellular experiments showed that both celastrol and PA could bind MD2, and that celastrol could expel PA from cells. Finally, celastrol could reverse high fat diet caused hyperglycemia and obesity, and NF-kB activations. Taking together, we proved that celastrol could reverses PA-caused TLR4-MD2 activation-dependent insulin resistance via disrupting PA binding to MD2.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced insulin resistance that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and metabolism, insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

Effects of Fat and Acids on the Formation of Autolysosomes in the Livers from Yellow Catfish Pelteobagrus Fulvidraco.

The autophagy-lysosome pathway, which involves many crucial genes and proteins, plays crucial roles in the maintenance of intracellular homeostasis by the degradation of damaged components. At present, some of these genes and proteins have been identified but their specific functions are largely unknown. This study was performed to clone and characterize the full-length cDNA sequences of nine key autolysosome-related genes ( and ) from yellow catfish Pelteobagrus fulvidraco. The expression of these genes and the transcriptional responses to a high-fat diet and acids (FAs) ( (PA) and oleic (OA)) were investigated. The mRNAs of these genes could be detected in heart, , muscle, spleen, brain, mesenteric adipose tissue, intestine, kidney and ovary, but varied with the tissues. In the , the mRNA levels of the nine autolysosome-related genes were lower in fish fed a high-fat diet than those fed the control, indicating that a high-fat diet inhibited formation of autolysosomes. (a saturated FA) significantly inhibited the formation of autolysosomes at 12 h, 24 h and 48 h incubation. In contrast, oleic (an unsaturated FA) significantly induced the formation of autolysosomes at 12 h, but inhibited them at 24 h. At 48 h, the effects of OA incubation on autolysosomes were OA concentration-dependent in primary hepatocytes of P. fulvidraco. The results of flow cytometry and laser confocal observations confirmed these results. PA and OA incubation also increased intracellular non-esterified (NEFA) concentration at 12 h, 24 h and 48 h, and influenced mRNA levels of binding protein () and transport protein 4 () which facilitate FA transport in primary hepatocytes of P. fulvidraco. The present study demonstrated the molecular characterization of the nine autolysosome-related genes and their transcriptional responses to fat and FAs in fish, which provides the basis for further exploring their regulatory mechanism in vertebrates.

Keyword: fatty liver

Palmitate and Stearate are Increased in the Plasma in a 6-OHDA Model of Parkinson\'s Disease.

Parkinson\'s disease (PD) is the second most common neurodegenerative disorder, without any widely available curative therapy. Metabolomics is a powerful tool which can be used to identify unexpected pathway-related disease progression and pathophysiological mechanisms. In this study, metabolomics in brain, plasma and was investigated in an experimental PD model, to discover small molecules that are associated with dopaminergic cell loss.Sprague Dawley (SD) rats were injected unilaterally with 6-hydroxydopamine (6-OHDA) or saline for the vehicle control group into the medial forebrain bundle (MFB) to induce loss of dopaminergic neurons in the substantia nigra pars compacta. Plasma, midbrain and samples were collected for metabolic profiling. Multivariate and univariate analyses revealed metabolites that were altered in the PD group.In plasma, ( = 3.72 × 10, FC = 1.81) and stearic ( = 3.84 × 10, FC = 2.15), were found to be increased in the PD group. ( = 3.5 × 10) and stearic ( = 2.7 × 10) correlated with test scores indicative of motor dysfunction. Monopalmitin ( = 4.8 × 10, FC = -11.7), monostearin ( = 3.72 × 10, FC = -15.1) and myo-inositol ( = 3.81 × 10, FC = -3.32), were reduced in the midbrain. The did not have altered levels of these molecules.Our results show that saturated free acids, their monoglycerides and myo-inositol metabolism in the midbrain and enteric circulation are associated with 6-OHDA-induced PD pathology.

Keyword: fatty liver

High vulnerability of the heart and to 3-hydroxypalmitic -induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic (3HPA), the long-chain hydroxyl (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca retention capacity and membrane potential in Ca -loaded mitochondria more markedly in the heart and the , with mild or no effects in the brain, supporting a higher susceptibility of the heart and the to the toxic effects of this . It is postulated that disruption of mitochondrial energy and Ca homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

A combination of plasma phospholipid acids and its association with incidence of type 2 diabetes: The EPIC-InterAct case-cohort study.

Combinations of multiple acids may influence cardiometabolic risk more than single acids. The association of a combination of acids with incident type 2 diabetes (T2D) has not been evaluated.We measured plasma phospholipid acids by gas chromatography in 27,296 adults, including 12,132 incident cases of T2D, over the follow-up period between baseline (1991-1998) and 31 December 2007 in 8 European countries in EPIC-InterAct, a nested case-cohort study. The first principal component derived by principal component analysis of 27 individual acids (mole percentage) was the main exposure (subsequently called the pattern score [FA-pattern score]). The FA-pattern score was partly characterised by high concentrations of linoleic , stearic , odd-chain acids, and very-long-chain saturated acids and low concentrations of γ-linolenic , , and long-chain monounsaturated acids, and it explained 16.1% of the overall variability of the 27 acids. Based on country-specific Prentice-weighted Cox regression and random-effects meta-analysis, the FA-pattern score was associated with lower incident T2D. Comparing the top to the bottom fifth of the score, the hazard ratio of incident T2D was 0.23 (95% CI 0.19-0.29) adjusted for potential confounders and 0.37 (95% CI 0.27-0.50) further adjusted for metabolic risk factors. The association changed little after adjustment for individual acids or subclasses. In cross-sectional analyses relating the FA-pattern score to metabolic, genetic, and dietary factors, the FA-pattern score was inversely associated with adiposity, triglycerides, enzymes, C-reactive protein, a genetic score representing insulin resistance, and dietary intakes of soft drinks and alcohol and was positively associated with high-density-lipoprotein cholesterol and intakes of polyunsaturated fat, dietary fibre, and coffee (p < 0.05 each). Limitations include potential measurement error in the acids and other model covariates and possible residual confounding.A combination of individual acids, characterised by high concentrations of linoleic , odd-chain acids, and very long-chain acids, was associated with lower incidence of T2D. The specific pattern may be influenced by metabolic, genetic, and dietary factors.

Keyword: fatty liver

Dual PPARα/γ agonist saroglitazar improves histopathology and biochemistry in experimental NASH models.

Non-alcoholic disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are common clinico-pathological conditions that affect millions of patients worldwide. In this study, the efficacy of saroglitazar, a novel PPARα/γ agonist, was assessed in models of NAFLD/NASH.HepG2 cells treated with (PA;0.75 mM) showed decreased expression of various antioxidant biomarkers (SOD1, SOD2, glutathione peroxidase and catalase) and increased expression of inflammatory markers (TNFα, IL1β and IL6). These effects were blocked by saroglitazar, pioglitazone and fenofibrate (all tested at 10μM concentration). Furthermore, these agents reversed PA-mediated changes in mitochondrial dysfunction, ATP production, NFkB phosphorylation and stellate cell activation in HepG2 and HepG2-LX2 Coculture studies. In mice with choline-deficient high-fat diet-induced NASH, saroglitazar reduced hepatic steatosis, inflammation, ballooning and prevented development of fibrosis. It also reduced serum alanine aminotransferase, aspartate aminotransferase and expression of inflammatory and fibrosis biomarkers. In this model, the reduction in the overall NAFLD activity score by saroglitazar (3\xa0mg/kg) was significantly more prominent than pioglitazone (25\xa0mg/kg) and fenofibrate (100\xa0mg/kg). Pioglitazone and fenofibrate did not show any improvement in steatosis, but partially improved inflammation and function. Antifibrotic effect of saroglitazar (4\xa0mg/kg) was also observed in carbon tetrachloride-induced fibrosis model.Saroglitazar, a dual PPARα/γ agonist with predominant PPARα activity, shows an overall improvement in NASH. The effects of saroglitazar appear better than pure PPARα agonist, fenofibrate and PPARγ agonist pioglitazone.© 2017 Cadila Healthcare Ltd., International Published by John Wiley & Sons Ltd.

Keyword: fatty liver

Strontium Alleviates Endoplasmic Reticulum Stress in a Nonalcoholic Disease Model.

The purpose of this study was to explore the effects of strontium on , and to clarify the possible mechanisms by which strontium improves nonalcoholic disease (NAFLD). We also evaluated how strontium affected the endoplasmic reticulum stress (ERS) pathways. We established an in vitro model of NAFLD using a human hepatocyte cell line (L02) treated with 0.2\u2009mM . The Sprague-Dawley rats were fed with a high-fat diet (HFD) to establish NAFLD model in vivo. After strontium treatment, the total cholesterol (TC), triglyceride (TG), and lipid deposition in L02 cells and tissues were determined. Strontium treatment suppressed intracellular TC and TG levels and lipid accumulation in L02 cells, and the effect of high concentrations of strontium were more obvious. Strontium significantly reduced the mRNA and protein expression of glucose-regulated protein 78 (GRP78), activating transcription factor 6 (ATF6), inositol requiring enzyme 1 (IRE1), SREBP cleavage activator protein (SCAP), sterol regulatory element binding protein 1c (SREBP-1c), and SREBP-2 in L02 cells. In HFD-fed rats, strontium treatment reduced serum TC, TG, and low density lipoprotein cholesterol (LDL-C) levels, concurrent with a decrease in hepatic lipid accumulation. Furthermore, strontium treatment reduced the expression of GRP78 and SREBP-2 protein in tissues. Overall, strontium alleviated hepatic steatosis by decreasing ERS-related protein expression in vivo and in vitro models. The results indicated that strontium has the potential to become a new therapy for the prevention and treatment of NAFLD.

Keyword: fatty liver

Individual acids in erythrocyte membranes are associated with several features of the metabolic syndrome in obese children.

Obesity leads to the clustering of cardiovascular (CV) risk factors and the metabolic syndrome (MetS) also in children and is often accompanied by non-alcoholic disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting insulin (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting insulin (r\u2009=\u20090.287). Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

Keyword: fatty liver

Glucose and lipid metabolism disorders in the chickens with dexamethasone-induced oxidative stress.

The purpose of this study was to investigate the effects of long-term treatment with dexamethasone (DEX) on the antioxidation and nutrition metabolism in broiler chickens. Broilers were placed on a high-nutrient diet for 41\xa0days, and half were given orally DEX-supplemented water at 20\xa0mg/L every other day from 19 to 41\xa0days of age. DEX treatment downregulated superoxide dismutase activity as well as the mRNA expression of CuZn-superoxide dismutase and glutathione peroxidase with a decrease in GSH/GSSG ratio and an increase in malondialdehyde level in the of broilers. DEX treatment aggravated oxidative damage in the and, therefore, increased the sensitivity of broilers to ascites syndrome with higher mortality and reduced growth performance. Serum metabolomics analysis showed that DEX treatment significantly increased the levels of glucose, intermediates in protein metabolism (valine, proline, serine, threonine and urea) and lipid metabolism-related products (, stearic and cholesterol) while decreasing the levels of β-hydroxy butyric , succinic and malic , demonstrating that DEX treatment inhibited the Krebs cycle and the oxidation of acids, and promoted the de novo synthesis of acids as well as protein decomposition in the of broilers. Additionally, detection of metabolism-related enzymes\xa0revealed\xa0that DEX treatment inhibited glycolysis and promoted glycogen decomposition. In summary, DEX treatment resulted in oxidative stress and glucose and lipid\xa0metabolism disorders in the broilers.© 2017 Blackwell Verlag GmbH.

Keyword: fatty liver

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the , and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated inflammation.

Keyword: fatty liver

[Effects of sera of rats fed with tablets on endoplasmic reticulum stress in a HepG2 cell model of nonalcoholic disease].

To investigate the effects of sera from rats fed with tablets (HGT) on endoplasmic reticulum (ER) stress in a steatotic hepatocyte model of free acids (FFAs)-induced nonalcoholic disease (NAFLD) and explore the possible mechanism.FFAs prepared by mixing oleic and at the ratio of 2:1. HepG2 cells were treated with the sera from rats fed with low-, moderate-or high-dose HGT (HGT sera) or sera of rats fed with fenofibrate (fenofibrate sera), followed by treatment with 1 mmol/L FFAs for 24 h to induce hepatic steatosis. Oil red O staining was used to observe the distribution of lipid droplets in the cells. The biochemical parameters including triglyceride (TG), lactated hydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using a commercial kit. The morphological changes of the ER in the cells were observed using transmission electron microscopy. The protein/mRNA expressions of ER stress-related signal molecules including GRP78, PERK, p-PERK, ATF6, ATF4, CASPASE-12, CHOP, XBP-1, PKC, and p-PKC-δ were detected using Western blotting and/or quantitative real-time PCR (qRT-PCR). The changes in the protein expressions of GRP78, p-PERK, CASPASE-12 and CHOP were also detected in cells with transient transfection of PKC-δ siRNA for PKC-δ knockdown.Compared with the control cells, the cells treated with FFAs showed significantly increased levels of TG, AST, and ALT ( < 0.05). Compared with FFAs-treated cells, the cells pretreated with HGT sera or fenofibrate sera all showed significantly decreased TG, AST and ALT levels ( < 0.05), reduced accumulation of the lipid droplets ( < 0.05), and lowered protein or mRNA expression levels of GRP78, p-PERK, ATF6, ATF4, CHOP, CASPASE-12, XBP-1 and p-PKC-δ ( < 0.05). PKC-δ knockdown caused significantly reduced protein expressions of GRP78, p-PERK, CASPASE-12 and CHOP in the cells with FFA-induced hepatic steatosis ( < 0.001); treatment with high-dose HGT serum more significantly reduced the expressions of GRP78 ( < 0.001) and P-PERK ( < 0.01) in FFAs-induced cells with PKC-δ knockdown.HGT serum can effectively prevent FFAs-induced steatosis in HepG2 cells by alleviating ER stress, in which PKC-δ may act as an important target.

Keyword: fatty liver

Human Milk and Donkey Milk, Compared to Cow Milk, Reduce Inflammatory Mediators and Modulate Glucose and Lipid Metabolism, Acting on Mitochondrial Function and Oleylethanolamide Levels in Rat Skeletal Muscle.

Milk from various species differs in nutrient composition. In particular, human milk (HM) and donkey milk (DM) are characterized by a relative high level of triacylglycerol enriched in in sn-2 position. These dietary fats seem to exert beneficial nutritional properties through N-acylethanolamine tissue modulation. The aim of this study is to compare the effects of cow milk (CM), DM, and HM on inflammation and glucose and lipid metabolism, focusing on mitochondrial function, efficiency, and dynamics in skeletal muscle, which is the major determinant of resting metabolic rate. Moreover, we also evaluated the levels of endocannabinoids and N-acylethanolamines in and skeletal muscle, since tissue profiles can be modulated by nutrient intervention. To this aim, rats were fed with CM, DM, or HM for 4 weeks. Then, glucose tolerance and insulin resistance were analyzed. Pro-inflammatory and anti-inflammatory cytokines were evaluated in serum and skeletal muscle. Skeletal muscle was also processed to estimate mitochondrial function, efficiency, and dynamics, oxidative stress, and antioxidant/detoxifying enzyme activities. profiles, endocannabinoids, and N-acylethanolamine congeners were determined in and skeletal muscle tissue. We demonstrated that DM or HM administration reducing inflammation status, improves glucose disposal and insulin resistance and reduces lipid accumulation in skeletal muscle. Moreover, HM or DM administration increases redox status, and mitochondrial uncoupling, affecting mitochondrial dynamics in the skeletal muscle. Interestingly, HM and DM supplementation increase and muscle levels of the N-oleoylethanolamine (OEA), a key regulator of lipid metabolism and inflammation. HM and DM have a healthy nutritional effect, acting on inflammatory factors and glucose and lipid metabolism. This beneficial effect is associated to a modulation of mitochondrial function, efficiency, and dynamics and to an increase of OEA levels in skeletal muscle.

Keyword: fatty liver

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and -to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the . GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

MiR-181b regulates steatosis in nonalcoholic disease via targeting SIRT1.

Non-alcoholic diseases (NAFLD) is one of the leading cause of chronic diseases in the world. However, the pathogenesis of NAFLD is still unclear. Emerging studies have demonstrated that microRNAs (miRs) are profoundly involved in NAFLD and related metabolic diseases. Here, we investigated the mechanisms by which miR-181b influences NAFLD via direct targeting SIRT1. The expression of miR181b was up-regulated while SIRT1 was down-regulated in both human NAFLD patients and high fat diet (HFD) induced NAFDL mice model. And (PA) treatment increased the miR-181b expression while decreased SIRT1 expression in HepG2 cells. Further, we identified that SIRT1 is a direct downstream target of miR-181b. Ectopic expression of miR-181b significantly repressed the 3\'-UTR reporter activities of SIRT1 in a dose-dependent manner, while the effect of miR-181b was interrupted when the binding site of miR-181b within the SIRT1 3\'-UTR was mutated. And overexpression of miR-181b reduced both the mRNA and protein levels of SIRT1 in HepG2 cells. We also found that inhibition of miR-181b expression alleviates hepatic steatosis both in\xa0vitro and in\xa0vivo. And the effect of miR-181b on steatosis was blocked by SIRT1 overexpression. Taken together, our data indicated that increased expression of miR-181b potentially contributes to altered lipid metabolism in NAFLD. Downregulation of miR-34a may be a therapeutic strategy against NAFLD by regulating its target SIRT1.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: fatty liver

Ellagic Suppresses the Oxidative Stress Induced by Dietary-Oxidized Tallow.

Dietary tallow was thermally oxidized at 180°C in an open fryer. The oxidized tallow (OT) and unoxidized tallow were characterized for oxidation parameters and composition using GC-MS. Tallow samples were fed to rabbits along with 50, 100, and 150\u2009mg/kg/day of ellagic (EA) for three weeks. Results revealed that the peroxide value (PV) and thiobarbituric reactive substances (TBARS) significantly increased, while radical scavenging activity (RSA) of the tallow decreased significantly with oxidation. GC-MS analysis showed eight acids in the tallow samples, where (48.5-49.7\u2009g/100\u2009g), linoleic (18.7-23.7\u2009g/100\u2009g), stearic (13.5-15.6\u2009g/100\u2009g), and margaric (6.32-6.42\u2009g/100\u2009g) were the major acids. Animal studies showed that oxidized tallow (OT) alone or in combination with EA significantly altered the body weight of the rabbits. Serum biochemical parameters and renal function tests were affected by OT and ameliorated by EA. The toxic effects of OT on haematological indices were minimized by EA. The supplementation of OT alone had significant effects on the structure and functions. The coadministration of EA reduced the toxic properties of OT on the , by increasing the antioxidant (GSH) system. The rabbit heart was also affected by the OT, which was ameliorated by EA supplementation. These results suggested that the supplementation of EA was beneficial against the OT-induced oxidative stress and may be considered for foods containing oxidized lipids.

Keyword: fatty liver

The effect of enterolactone on lipid precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on lipid fractions acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of acids were evaluated by Western Blot.Enterolactone modified acids composition in FFA, DAG and TAG fractions. In conjunction with lipid overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated acids in each of the investigated lipid fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Downregulation of sirtuin 3 by increases the oxidative stress, impairment of mitochondrial function, and apoptosis in cells.

Elevated levels of saturated acids show a strong cytotoxic effect in cells. Sirtuin 3 (SIRT3), a mitochondrially localized member of NAD -dependent deacetylase has been shown to protect hepatocytes against the oxidative stress. The role of SIRT3 on the cytotoxicity caused by acids in cells is not fully understood. The aim of this study was to evaluate the expression level of SIRT3, oxidative stress, and mitochondrial impairments in human hepatoma HepG2 cells exposed to (PA). Our results showed that PA treatment caused the deposition of lipid droplets and resulted in an increased expression of tumor necrosis factor-α in a dose-dependent manner. Excessive accumulation of PA induces the reactive oxygen species formation and apoptosis while dissipating the mitochondrial transmembrane potential. The level of SIRT3 expression in both nuclear and mitochondrial fractions in HepG2 cells was decreased with the increase in PA concentrations. However, in the cytosolic fraction, the SIRT3 was undetectable. In conclusion, our results showed that PA caused an increase in inflammation and oxidative stress in HepG2 cells. The exposure of PA also resulted in the decline in transmembrane potential and an increase in apoptosis. The underexpression of nuclear and mitochondrial SIRT3 by PA suggests that the PA target the process that regulates the stress-related gene expression and mitochondrial functions.© 2019 Wiley Periodicals, Inc.

Keyword: fatty liver

[The person in philogenesis is not (omnivores), but the herbivores with the carnivores past and the fuzzy future. Biological function of trophology (nutrition) in ontogenesis.]

According to the phylogenetic theory of general pathology, seven biological functions have been formed over billions of years. 1. biological function of trophology, nutrition; 2. homeostasis function; 3. biological function of endoecology; 4. function of adaptation; 5. function of the continuation of the species; 6. function of locomotion and 7. cognitive biological function, including intelligence. Millions of years in life consistently in the waters of several oceans, all the ancestors of man were carnivorous (Carnivores), fish-eating mammals. When the ocean retreated and the carnivorous (fish-eating) were on land, each individual privatized a "piece" of the ocean. Animals transformed it ito a pool of intercellular medium in vivo. The biological role of the late in the phylogeny of insulin is the formation of new biological functions in vivo. The action of insulin has transformed the carnivorous (fish-eating) ocean into herbivorous (Herbivores) species on land. There was it by synthesis in vivo from exogenous glucose of acids (FA). Regulatory action of insulin was the directed conversion of exogenous glucose into ω-6 C18: 1 cis-oleic FA. Insulin late in phylogeny expressed the synthesis of new, conjugated enzymes: it is palmitoyl-CoA-elongase and stearyl-CoAdesaturase. Two enzymes synthesized FAs along the way: synthesized in situ de novo, from exogenous glucose, C16: 0 → C18: 0 stearic → ω-6 C18: 1 cis-oleic without accumulation of stearic FA. Insulin is not converted into an oleic FA exogenous from carnivorous food. On land, the action of insulin transformed the species Homo sapiens, into a herbivore, but with carnivorous, fish-eating, past. The idea of a person as omnivorous (Omnivor) - nonsense; such forms of nature did not form. Violation of the function of nutrition, the biological reaction of exotrophy (external nutrition), is the etiological and pathogenetic basis of the seven metabolic pandemics, the diseases of civilization. 1. Atherosclerosis and atheromatosis; 2. metabolic arterial hypertension; 3. metabolic syndrome; 4. obesity; 5. syndrome of insulin resistance; 6. non-alcoholic disease and 7. endogenous hyperuricemia. The primary prevention of metabolic pandemics in the biological function of nutrition, in the biological reactions of exo-and endotrophy, will allow us to understand the theoretical bases and implementation of preventive actions that will determine the characteristics of nutrition in the future.

Keyword: fatty liver

A Pathophysiological Model of Non-Alcoholic Disease Using Precision-Cut Slices.

Non-alcoholic disease (NAFLD) is a common disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut slices (PCLSs) were used in this study. PCLSs, prepared from tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: fatty liver

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Reporting temporal fluctuations of hepatic C16 and C18 acids during late gestation and early lactation in dromedary camel.

Based on current knowledge, C16 and C18 acids (FA) are considered the most functional FA in hepatic metabolism. Although these FAs have been satisfyingly investigated in cattle, other species such as camel have been neglected. For this reason, the current study was designed to scrutinize changing patterns of C16 and C18 FAs in 10 dromedary camels from the last 2\xa0months of gestation to the first\xa0months of lactation. Camels were grazed on natural pasture and supplemented with a balanced ration. biopsies were obtained through blind biopsy technique at about 60, 45, 30, and 15-day antepartum (AP), and at 3, 15, 30, 45, and 60 post-partum (PP). Data were analyzed by the ANOVA procedure of SPSS with repeated measurements. From 15-day AP, saturated FA content of the declined (P\u2009<\u20090.01) and 15-day PP reached its peak (P\u2009=\u20090.02). At 30-day PP it went down (P\u2009<\u20090.01), and re-elevated at 45-day PP (P\u2009<\u20090.01) but remained at a steady state for the duration of the study. Mono-unsaturated and polyunsaturated FA content of hepatic tissue were constant throughout AP, albeit observed to peak at 15-day AP compared with 45 (P\u2009=\u20090.04) and 30-day AP (P\u2009<\u20090.01) for mono-unsaturated FAs, and with 60-, 45-, and 30-day AP (P\u2009≤\u20090.01) for polyunsaturated FAs. The content of the reached a nadir at 30-day AP (P\u2009<\u20090.01), increased sharply (P\u2009<\u20090.01) at the next sampling time-point, and had a trend to escalate until 3-day PP. Palmitoleic levels were unchanged from 60- to 30-day AP, decreased at 15 AP and 3-day PP, increased at 15-day PP, then remained constant until the end of the study period (P\u2009≤\u20090.04). Stearic content started to grow at 15-day AP and reached its peak at 15-day PP (P\u2009<\u20090.01). At 30-day PP, stearic level in dropped abruptly (P\u2009<\u20090.01), then intensified at 45-day PP and did not change after; hepatic content of stearic was lower during AP compared with PP time-points. Other C18 FAs changed significantly during the study period. These results suggest that parturition could have a profound effect on FA composition and other metabolites in camel . Further research is required to establish the metabolic mechanism behind these changes.

Keyword: fatty liver

GLP-1 Elicits an Intrinsic Gut- Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN ( synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut- regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: fatty liver

Novel hypolipidemic conjugates of and bile with lysine for linkage.

Novel -bile conjugates (1a-1k) were designed and synthesized by coupling of the acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve -specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high-fat diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced damage.

Keyword: fatty liver

Human plasma retinol-binding protein (RBP4) is also a -binding protein.

RBP4 (plasma retinol-binding protein) is the 21\u202fkDa transporter of all-trans retinol that circulates in plasma as a moderately tight 1:1 molar complex of the vitamin with the protein. RBP4 is primarily synthesized in the but is also produced by adipose tissue and circulates bound to a larger protein, transthyretin, TTR, that serves to increase its molecular mass and thus avoid its elimination by glomerular filtration. This paper reports the high resolution three-dimensional structures of human RBP4 naturally lacking bound retinol purified from plasma, urine and amniotic fluid. In all these crystals we found a molecule bound in the hydrophobic ligand-binding site, a result confirmed by mass spectrometry measurements. In addition we also report the 1.5\u202fÅ resolution structures of human holo-RBP4 and of the protein saturated with and lauric and discuss the interaction of the acids and retinol with the protein.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Vildagliptin Can Alleviate Endoplasmic Reticulum Stress in the Induced by a High Fat Diet.

We investigated whether a DDP-4 inhibitor, vildagliptin, alleviated ER stress induced by a high fat diet and improved hepatic lipid deposition. C57BL/6 mice received standard chow diet (CD), high fat diet (HFD), and HFD administered with vildagliptin (50\u2009mg/Kg) (V-HFD). After administration for 12 weeks, serum alanine aminotransferase, glucose, cholesterol, triglyceride, and insulin levels were analyzed. Samples of underwent histological examination and transmission electron microscopy, real-time PCR for gene expression levels, and western blots for protein expression levels. ER stress was induced in HepG2 cells with and the effects of vildagliptin were investigated. HFD mice showed increased weight/body weight (20.27%) and triglycerides (314.75%) compared to CD mice, but these decreased by 9.27% and 21.83%, respectively, in V-HFD mice. In the , HFD induced the expression of ER stress indicators significantly, which were obviously decreased by vildagliptin. In vitro, the expressions of molecular indicators of ER stress were reduced in HepG2 when vildagliptin was administered. Vildagliptin alleviates hepatic ER stress in a mouse high fat diet model. In HepG2 cells, vildagliptin directly reduced ER stress. Therefore, vildagliptin may be a potential agent for nonalcoholic disease.

Keyword: fatty liver

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved -body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: fatty liver

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Metabolism Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic disease (NAFLD) has become the most common cause of chronic disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic metabolism. DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Diseases.

Keyword: fatty liver

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: fatty liver

Ganoderma Lucidum Polysaccharide Peptide Alleviates Hepatoteatosis via Modulating Bile Metabolism Dependent on FXR-SHP/FGF.

Non-alcoholic disease (NAFLD) encompasses a series of pathologic changes ranging from steatosis to steatohepatitis, which may progress to cirrhosis and hepatocellular carcinoma. The purpose of this study was to determine whether ganoderma lucidum polysaccharide peptide (GLPP) has therapeutic effect on NAFLD.Ob/ ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic pathways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blot. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD.GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, dysfunction and insulin resistance. Pathways of glycerophospholipid metabolism, metabolism and primary bile biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1, CYP8B1, FXR, SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepatocytes induced by oleic and .GLPP significantly improves NAFLD via regulating bile synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits synthesis, indicating that GLPP might be developed as a therapeutic drug for NAFLD.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Impact of Brain Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and energy balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with signaling. In humans circulating saturated acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain signaling and its contemporaneous effect on function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fatty liver

Dietary lipid requirement of lemon fin barb hybrid.

The present study has been conducted to determine the optimal dietary requirement lipid of lemon fin barb hybrid fingerlings. Five isonitrogenous diets (35% protein) with graded lipid levels of 0%, 4%, 6%, 8% and 12% were fed to 225 lemon fin barb hybrid fingerlings (4.3 ? 0.5 g), which were randomly and equally distributed into twelve 60 l aquaria. The diets were randomly assigned to the aquaria in triplicates, and the feeding experiment was conducted for 10 weeks. Fish were fed at 5% body weight and the feeding ration was adjusted fortnightly. The weight gain, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) of fish fed 4% dietary lipid were significantly superior (P<0.05) than those fed with other diets, except for 6% lipid. Except for whole body moisture content, there were significant differences (P<0.05) between whole body proximate composition and nutrient retentions of fish fed with different test diets. The results showed that was the main saturated (SFA), oleic was the predominant MUFA while arachidic and docosahexanoic were the major PUFAs in the muscle and tissue of fish. The n-3/n-6 ratio ranged from 0.66 to 2.65. From the results of this study, a dietary lipid of 4% was recommended for lemon fin barb hybrid. ?

Keyword: fatty liver

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic disease and nonalcoholic disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the , as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the accumulation. In NAFLD, QWTX attenuated steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.Copyright © 2019. Published by Elsevier B.V.

Keyword: fatty liver

[Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: fatty liver

In Vitro Monitoring of the Mitochondrial Beta-Oxidation Flux of and Investigation of Its Pharmacological Alteration by Therapeutics.

The present study was designed to validate the functional assay that enables rapid screening of therapeutic candidates for their effect on mitochondrial oxidation.The two whole-cell systems (tissue homogenates and hepatocytes) have been evaluated to monitor the total beta-oxidation flux of physiologically important H- by measurement of tritiated water enrichment in incubations using UPLC coupled on-line to radioactivity monitoring and mass spectrometry.Our results with several known inhibitors of oxidation showed that this simple assay could correctly predict a potential in alteration of mitochondrial function by drug candidates. Since the beta-oxidation of takes place almost exclusively in mitochondria of human hepatocytes, this model can be also utilized to distinguish between the mitochondrial and peroxisomal routes of this essential metabolic pathway in some cases.The present work offers a new in vitro screen of changes in mitochondrial beta-oxidation by xenobiotics as well as a model to study the mechanism of this pathway.

Keyword: fatty liver

Cypate and Cypate-Glucosamine as Near-Infrared Fluorescent Probes for In Vivo Tumor Imaging.

Near-infrared (NIR) imaging is a promising technique for use as a noninvasive and sensitive diagnostic tool. Although the NIR fluorescently labeled glucose analog glucosamine (cypate-glucosamine) has applications in preclinical imaging, the transport pathways and fate of this probe in tissues remain unaddressed. Here, we have synthesized and characterized cypate and cypate-glucosamine conjugate (cy-2-glu), and investigated the probable transport pathways of these probes in vitro and in vivo. We compared uptake of the probes in the presence and absence of excess d-glucose, "saturated cypate" and in two normal-cancer cell line pairs: lung cancer (A549)-normal (MRC9) and prostate cancer (DU145)-normal (BPH). Breast cancer (MDA-MB-231) and cancer (HepG2) cell lines were also examined. Results support use of the glucose transport pathway by cy-2-glu and transport pathway by cypate. Mass spectrometry data on the in vitro extracts revealed deamidation of cy-2-glu in prostate and cells, suggesting release of glucosamine. In vivo biodistribution studies in mice engrafted with breast tumors showed a distinct accumulation of cy-2-glu in and tumors, and to a lesser extent in kidneys and spleen. A negligible accumulation of cypate alone in tumors was observed. Analysis of urine extracts revealed renal excretion of the cy-2-glu probe in the form of free cypate, indicating deamidation of cy-2-glu in tissues. Thus, investigation of the metabolic pathways used by NIR probes such as cy-2-glu advances their use in the detection and monitoring of tumor progression in preclinical animal studies.Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: fatty liver

Thioacetamide potentiates high cholesterol and high fat diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH.

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Prepartum supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb and adipose tissue (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and and adipose tissue collected for FA and gene expression analysis. concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that lipogenesis decreased, and lipolysis increased in lamb with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that lipogenesis decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not , to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: fatty liver

Aberrant hepatic lipid storage and metabolism in canine portosystemic shunts.

Non-alcoholic disease (NAFLD) is a poorly understood multifactorial pandemic disorder. One of the hallmarks of NAFLD, hepatic steatosis, is a common feature in canine congenital portosystemic shunts. The aim of this study was to gain detailed insight into the pathogenesis of steatosis in this large animal model. Hepatic lipid accumulation, gene-expression analysis and HPLC-MS of neutral lipids and phospholipids in extrahepatic (EHPSS) and intrahepatic portosystemic shunts (IHPSS) was compared to healthy control dogs. organoids of diseased dogs and healthy control dogs were incubated with - and oleic-, and lipid accumulation was quantified using LD540. In histological slides of shunt livers, a 12-fold increase of lipid content was detected compared to the control dogs (EHPSS P<0.01; IHPSS P = 0.042). Involvement of lipid-related genes to steatosis in portosystemic shunting was corroborated using gene-expression profiling. Lipid analysis demonstrated different triglyceride composition and a shift towards short chain and omega-3 acids in shunt versus healthy dogs, with no difference in lipid species composition between shunt types. All organoids showed a similar increase in triacylglycerols after free acids enrichment. This study demonstrates that steatosis is probably secondary to canine portosystemic shunts. Unravelling the pathogenesis of this hepatic steatosis might contribute to a better understanding of steatosis in NAFLD.

Keyword: fatty liver

Synthesis and evaluation of Ga labeled for cardiac metabolic imaging.

This work evaluates the potential of a Ga labeled long chain 16C for cardiac metabolic imaging. For radiolabeling with Ga, hexadecanedioic was coupled with the chelator p-NH-Bn-NOTA. Under the optimized conditions, NOTA-hexadecanoic could be radiolabeled with Ga in ≥95% yields. In biodistribution studies carried out in Swiss mice, Ga-NOTA-hexadecanoic showed low myocardial uptake at 2\u202fmin p.i. (3.7\u202f±\u202f1.3%ID/g). While Ga-NOTA-hexadecanoic cleared rapidly from non-target organs such as blood, lungs, intestine and kidney, wash out from was slow. Radio-HPLC analyses of myocardial extracts of rats injected with Ga-NOTA-hexadecanoic confirmed its metabolic transformation in the myocardium.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Berberine attenuates nonalcoholic hepatic steatosis through the AMPK-SREBP-1c-SCD1 pathway.

Berberine (BBR), a natural compound extracted from Chinese herb, has been shown to effectively attenuate nonalcoholic disease (NAFLD) in clinic. However, the mechanism underlying the effect of BBR is not fully understood. Stearyl-coenzyme A desaturase 1 (SCD1) mediates lipid metabolism in . Therefore, we hypothesized that SCD1 mediated the beneficial effect of BBR on NAFLD.The expression of SCD1 was measured in the of NAFLD patients and ob/ob mice. The effect of BBR on NAFLD was evaluated in C57BL/6\u202fJ mice on high fat diet (HFD). The effect of BBR was also investigated in HepG2 and AML12\u202fcells exposed to high glucose and . Oil red O staining was performed to detect triglyceride (TG) level. Quantitative real-time polymerase chain reaction and Western blot were used to detect the messenger ribonucleic (mRNA) and protein expression of target genes. The activity of SCD1 promoter was measured by dual-luciferase reporter assay.The expression of SCD1 was increased in the of NAFLD patients and ob/ob mice. BBR reduced hepatic TG accumulation and decreased the expressions of hepatic SCD1 and other TG synthesis related genes both in vivo and in vitro. Knockdown of SCD1 expression mimicked the effect of BBR decreasing TG level in steatotic hepatocytes, whereas overexpression of SCD1 attenuated the effect of BBR. Mechanistically, BBR promoted the phosphorylation of AMP-activated protein kinase (AMPK) and sterol regulatory element-binding protein-1c (SREBP-1c) in HepG2 cells and the of HFD-fed mice. Activation of the AMPK-SREBP-1c pathway and sterol regulatory element (SRE) motif in SCD1 promoter (-920/-550) was responsible for the BBR-induced suppression of SCD1.BBR reduces TG synthesis and attenuates hepatic steatosis through the activation of AMPK-SREBP-1c-SCD1 pathway.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high energy value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on lipid metabolism are still poorly understood. Therefore, this study evaluated the (FA) profile and the effects of baru oil on and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a lipid emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and . The results demonstrated that animals developed marked hypercholesterolemia, steatosis, and increased lipid peroxidation in the aorta. Treatment with baru oil attenuated lipid peroxidation and drastically reduced damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: fatty liver

Inhibitory effect of 17β‑estradiol on triglyceride synthesis in skeletal muscle cells is dependent on ESR1 and not ESR2.

The present study aimed to investigate the inhibitory effects and the mechanisms underlying 17β‑estradiol (E2) effects on triglyceride synthesis and insulin resistance in skeletal muscle tissues and cells. Ovariectomy (OVX) was performed on 6‑month‑old female rats treated with or without E2. Subsequently, various serum biochemical markers were measured. Additionally, pathological alterations of the uterus, and skeletal muscle were analyzed, and the content of triglycerides (TG) in muscle was detected. Differentiated myotubes formed by C2C12 cells were treated with (PA) or pretreated with E2, estrogen receptor (ESR)\xa01 agonist propylpyrazoletriol (PPT) and ESR2 agonist diarylpropionitrile (DPN). Subsequently, the mRNA or protein expression levels of ESR1/2, peroxisome proliferator activated receptor α (PPARα), CD36 molecule (CD36), synthase (FASN), perilipin 2 (PLIN2), phosphorylated acetyl‑CoA carboxylase α (p‑ACACA), p‑AKT serine/threonine kinase (p‑AKT) and p‑mitogen‑activated protein kinase\xa08 (p‑MAPK8) were analyzed in skeletal muscle or in C2C12 cells by reverse transcription‑semi‑quantitative polymerase chain reaction and western blotting. The present results suggested that treatment with E2 inhibited OVX‑induced body weight gain, TG accumulation and insulin resistance. The protein or mRNA expression levels of ESR1, CD36, PPARα, p‑ACACA and p‑AKT were decreased, whereas the protein or mRNA expression levels of ESR2, PLIN2, FASN and p‑MAPK8 were increased in the OVX group. Of note, treatment with E2 restored the expression levels of the aforementioned factors. In C2C12 cells, treatment with E2 or PPT reversed the alterations induced by treatment with PA. In contrast, pretreatment with DPN did not influence the effect of PA. Collectively, E2 was able to interact with ESR1, thus activating the CD36‑PPARα pathway, decreasing the level of TG in the muscles and improving insulin resistance in skeletal muscles and C2C12 cells.

Keyword: fatty liver

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: fatty liver

A unifying mathematical model of lipid droplet metabolism reveals key molecular players in the development of hepatic steatosis.

The responds to elevated plasma concentrations of free acids (FFAs) with an enhanced uptake of FFAs and their esterification to triacylglycerol (TAG). On the long term, this may result in massive hepatic TAG accumulation called steatosis hepatitis. In hepatocytes, the poor water-soluble TAG is packed in specialized organelles: Lipid droplets (LDs) serving as transient cellular deposit and lipoproteins (LPs) transporting TAG and cholesterol esters to extra-hepatic tissues. The dynamics of these organelles is controlled by a variety of regulatory surface proteins (RSPs). Assembly and export of VLDLs are mainly regulated by the microsomal transfer protein (MTP) and apoprotein B100. Formation and lipolysis of LDs are regulated by several RSPs. The best studied regulators belong to the PAT (Perilipin/Adipophilin/TIP47) and CIDE families. Knockdown or overexpression of SRPs may significantly affect the total number and size distribution of LDs. Intriguingly, a large cell-to-cell heterogeneity with respect to the number and size of LDs has been found in various cell types including hepatocytes. These findings suggest that the extent of cellular lipid accumulation is determined not only by the imbalance between lipid supply and utilization but also by variations in the expression of RSPs and metabolic enzymes. To better understand the relative regulatory impact of individual processes involved in the cellular TAG turnover, we developed a comprehensive kinetic model encompassing the pathways of the and triglyceride metabolism and the main molecular processes governing the dynamics of LDs. The model was parametrized such that a large number of experimental in vitro and in vivo findings are correctly recapitulated. A control analysis of the model revealed that variations in the activity of FFA uptake, diacylglycerol acyltransferase (DGAT) 2, and adipose triglyceride lipase (ATGL) have the strongest influence on the cellular TAG level. We used the model to simulate LD size distributions in human hepatoma cells and hepatocytes exposed to a challenge with FFAs. A random fold change by a factor of about two in the activity of RSPs was sufficient to reproduce the large diversity of droplet size distributions observed in individual cells. Under the premise that the same extent of variability of RSPs holds for the intact organ, our model predicts variations in the TAG content of individual hepatocytes by a factor of about 3-6 depending on the nutritional regime. Taken together, our modeling approach integrates numerous experimental findings on individual processes in the cellular TAG metabolism and LD dynamics metabolism to a consistent state-of-the-art dynamic network model that can be used to study how changes in the external conditions or systemic parameters will affect the TAG content of hepatocytes.© 2017 Federation of European Biochemical Societies.

Keyword: fatty liver

Increased plasma oleoylethanolamide and palmitoleoylethanolamide levels correlate with inflammatory changes in alcohol binge drinkers: the case of HMGB1 in women.

Alcohol binge drinking is a heavy pattern of alcohol consumption increasingly used by young people. In a previous study, we reported that young drinkers with a 2-year history of binge alcohol consumption had an overactivation of the innate immune system and peripheral inflammation when compared with controls. In the present study, we measured several biolipids that are derivatives belonging to the acylethanolamide or 2-acylglycerol families in the plasma of the same subjects (n\xa0=\xa042; 20 men and 22 women). We found that during abstinence, alcohol binge drinkers had elevated plasma levels of oleoylethanolamide, palmitoleoylethanolamide, arachidonoylethanolamide, dihomo-γ-linolenoyl ethanolamide and linoleoyl ethanolamide, which positively correlated with changes in the mRNA expression of key inflammatory markers in peripheral blood mononuclear cells, such as toll-like receptors (TLR4), pro-inflammatory cytokines/chemokines interleukin-1 beta, interleukin-6 and monocyte chemoattractant protein-1, and cyclooxygenase-2. Additionally, plasma oleoylethanolamide positively correlated with plasma levels of high mobility group box-1, which is a danger-associated molecular pattern and an endogenous TLR4 agonist, specifically in female alcohol binge drinkers. No changes were observed in 2-acylglycerols in alcohol binge drinkers, although sex-related differences in these bioactive lipids as well as in palmitoleoylethanolamide and docosatetraenoylethanolamide levels were detected. These results extend the previous clinical findings observed in patients diagnosed with long-term alcohol use disorder to young users and suggest a prominent role for these lipids in the response to acute alcohol exposure.© 2017 Society for the Study of Addiction.

Keyword: fatty liver

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

Inflammation markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free acids (FFAs) are known to cause damage in various tissues by inducing inflammation. In this study, we investigated whether a FFA (palmitate) induces inflammation in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive oxygen species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including diabetes.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Hepatitis C Virus Infection Increases c-Jun N-Terminal Kinase (JNK) Phosphorylation and Accentuates Hepatocyte Lipoapoptosis.

BACKGROUND Hepatitis C virus (HCV) infection and metabolic diseases including nonalcoholic steatohepatitis (NASH) exhibit a complex interplay. Although free -mediated apoptosis is a prominent feature of NASH, the impact of HCV infection on hepatocyte lipotoxicity has remained largely unexplored. The study aimed at identifying whether infection by HCV affected the apoptotic pathway in hepatocytes during assault. MATERIAL AND METHODS OR6 cells, which are derived from human hepatocellular carcinoma Huh-7 cells and harbor a full-length HCV RNA genome replication system, were treated with palmitate. Apoptosis was examined by 4\',6-diamidino-2-phenylindole staining. Activation and expression of JNK, Bim, cIAP-1, and Mcl-1 were examined by immunoblotting. mRNA expression of CHOP, a major player in endoplasmic reticulum stress-mediated apoptosis, was assessed by real-time PCR. RESULTS Palmitate-induced hepatocyte apoptosis was significantly enhanced in OR6 cells compared to cured cells, in which the HCV genome had been eradicated by treatment with interferon-α. Although basal expression of CHOP mRNA was enhanced in OR6 cells compared to cured cells, it was similarly upregulated in both cell lines following palmitate treatment. Notably, palmitate-induced JNK phosphorylation was accentuated in OR6 cells compared to cured cells. Inhibition of JNK with SP600125 attenuated palmitate-induced apoptosis. Palmitate-mediated upregulation of BH3-only protein Bim, which acts downstream of JNK, was also enhanced in OR6 cells compared to cured cells. In contrast, Mcl-1 and cIAP-1 were equally reduced in OR6 cells and cured cells following palmitate treatment. CONCLUSIONS These findings suggest that during lipoapoptosis, HCV infection may enhance hepatocyte toxicity by increasing JNK phosphorylation.

Keyword: fatty liver

TRIF-dependent Toll-like receptor signaling suppresses transcription in hepatocytes and prevents diet-induced hepatic steatosis.

Nonalcoholic disease (NAFLD) includes a spectrum of diseases that ranges in severity from hepatic steatosis to steatohepatitis, the latter of which is a major predisposing factor for cirrhosis and cancer. Toll-like receptor (TLR) signaling, which is critical for innate immunity, is generally believed to aggravate disease progression by inducing inflammation. Unexpectedly, we found that deficiency in TIR domain-containing adaptor-inducing interferon-β (TRIF), a cytosolic adaptor that transduces some TLR signals, worsened hepatic steatosis induced by a high-fat diet (HFD) and that such exacerbation was independent of myeloid cells. The aggravated steatosis in mice was due to the increased hepatocyte transcription of the gene encoding stearoyl-coenzyme A (CoA) desaturase 1 (SCD1), the rate-limiting enzyme for lipogenesis. Activation of the TRIF pathway by polyinosinic:polycytidylic [poly(I:C)] suppressed the increase in SCD1 abundance induced by or an HFD and subsequently prevented lipid accumulation in hepatocytes. Interferon regulatory factor 3 (IRF3), a transcriptional regulator downstream of TRIF, acted as a transcriptional suppressor by directly binding to the promoter. These results suggest an unconventional metabolic function for TLR/TRIF signaling that should be taken into consideration when seeking to pharmacologically inhibit this pathway.Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Keyword: fatty liver

Pioglitazone Enhances Cytosolic Lipolysis, β-oxidation and Autophagy to Ameliorate Hepatic Steatosis.

Non-alcoholic disease closely contributes to the development of obesity and insulin resistance. Even though pioglitazone has been reported to effectively lessen hepatic steatosis in human studies, its molecular mechanism remains unclear. This study is designed to investigate the regulation of cytosolic lipolysis, β-oxidation and autophagy by pioglitazone in a mice model of high fat diet (HFD) and cell model incubated with . Our results revealed hepatic steatosis was apparently induced by HFD and it was significantly reversed by pioglitazone. The serum insulin and hepatic triglyceride content was significantly decreased by co-administered pioglitazone with HFD. Hepatic expression of cytosolic-lipolysis related proteins (ATGL, HSL), β-oxidation (CPT-1A) and autophagy-related proteins (ATG7, LC3, LAL) was significantly enhanced by pioglitazone. Knockdown PPARα/PPARγ in AML12 cells significantly and proportionally reduced the expressions of ATGL, CPT-1A and LC3II, which was induced by pioglitazone. Furthermore, facilitation of the autophagic flux by pioglitazone was obviously blocked by lysosomal inhibitor, leupeptin, to demonstrate accumulation of the LC3II and intracellular lipid in AML12 cells. Our results demonstrated that pioglitazone attenuating the hepatic steatosis may be mediated by enhancing cytosolic lipolysis, β-oxidation and autophagy in a PPARα and PPARγ dependent manner.

Keyword: fatty liver

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: fatty liver

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis (NASH) is characterized by lipid accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on NASH and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce NASH and fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in inflammation and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated lipid accumulation, inflammation and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents NASH and fibrosis via inhibition of oxidative stress and inflammation, highlighting polydatin as a potential therapeutic agent for prevention and treatment of NASH.

Keyword: fatty liver

GC-MS metabolomics reveals disturbed metabolic pathways in primary mouse hepatocytes exposed to subtoxic levels of 3,4-methylenedioxymethamphetamine (MDMA).

3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a well-known hepatotoxic drug. Although its toxicity has been thoroughly studied at high concentrations, there is still insufficient knowledge on possible alterations of cell function at subtoxic concentrations, which are in fact more representative concentrations of intoxication scenarios. In this study, a gas chromatography-mass spectrometry (GC-MS) metabolomics approach was used to investigate the metabolic changes in primary mouse hepatocytes (PMH) exposed to two subtoxic concentrations of MDMA (LC and LC) for 24\xa0h. Metabolomic profiling of both intracellular metabolites and volatile metabolites in the extracellular medium of PMH was performed. Multivariate analysis showed that the metabolic pattern of cells exposed to MDMA discriminates from the controls in a concentration-dependent manner. Exposure to LC MDMA induces a significant increase in some intracellular metabolites, including oleic and , and a decrease in glutamate, aspartate, 5-oxoproline, fumarate, malate, phosphoric , α-ketoglutarate and citrate. Extracellular metabolites such as acetophenone, formaldehyde, pivalic , glyoxal and 2-butanone were found significantly increased after exposure to MDMA, compared to controls, whereas 4-methylheptane, 2,4-dimethyl-1-heptene, nonanal, among others, were found significantly decreased. The panel of discriminatory metabolites is mainly involved in tricarboxylic (TCA) cycle, metabolism, glutamate metabolism, antioxidant defenses and possibly changes in the enzyme machinery. Overall, these results highlight the potential of the intra- and extracellular metabolome to study alterations triggered by subtoxic concentrations of MDMA in hepatic cell functions, which represents a more realistic appraisal of early toxicity events posed by exposure to this drug. In addition, these results also revealed some metabolites that may be used as potential biomarkers indicative of early events in the hepatotoxicity induced by MDMA.

Keyword: fatty liver

Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.

Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 diabetes mellitus (T2DM) and to determine the underlying mechanisms of these effects.A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with . Major signalling profiles involved in oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry.Nuciferine restored impaired glucose tolerance and insulin resistance in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed -induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation.Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.© 2018 The British Pharmacological Society.

Keyword: fatty liver

Free -induced histone acetyltransferase activity accelerates lipid accumulation in HepG2 cells.

Non-alcoholic disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and lipid metabolism in the have not been thoroughly investigated.Following oleic- (OPA)-induced lipid accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between lipid accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and lipid accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: fatty liver

Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured Hepatocytes.

A fluorescence microscopic method for characterizing size, quantity, and oxidation of lipid droplets (LDs) in HepG2 cells was developed. LDs were induced by (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and lipid peroxides. Each fatty increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2\'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.

Keyword: fatty liver

Alleviation of -induced endoplasmic reticulum stress by augmenter of regeneration through IP3R-controlled Ca release.

The aberrant release of Ca from the endoplasmic reticulum (ER) contributes to the onset of ER stress, which is closely related to the pathogenesis of non-alcoholic disease. We previously reported that augmenter of regeneration (ALR) alleviates ER stress and protects hepatocytes from lipotoxicity. However, the link between ALR protection and the suppression of ER stress remains unclear. In this study, we investigated whether the protection against steatosis afforded by ALR is related to its inhibition of calcium overflow from the ER to the mitochondria. The treatment of HepG2 cells with (PA) upregulated IP3R expression, triggering ER-luminal Ca release and inducing ER stress. However, in ALR-transfected (ALR-Tx) HepG2 cells, PA-induced cell injury was clearly alleviated compared with that in vector-Tx cells. After exposure to PA, IP3R expression was downregulated and ER stress was effectively inhibited in the ALR-Tx cells, and ER-Ca release and simultaneous mitochondrial Ca uptake were lower than those in vector-Tx cells. The knockdown of ALR expression with shRNA abolished the protective effects afforded by ALR transfection. PA treatment also suppressed the interaction between BCL-2 and IP3R in HepG2 cells, whereas this interaction was massively enhanced in the ALR-Tx cells, effectively reducing the IP3R-mediated ER-Ca release and thus mitochondrial Ca influx. Our results suggest that the inhibition of ER stress by ALR is related to the interruption of the interaction between BCL2 and IP3R, demonstrating a novel mechanism of ER stress resistance in ALR-Tx cells.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Low molecular weight fucoidan attenuates injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in diabetes-induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for obesity/diabetes-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Attenuation of -Induced Lipotoxicity by Chlorogenic through Activation of SIRT1 in Hepatocytes.

Saturated free acids (FFAs) induce hepatocyte lipotoxicity, wherein oxidative stress-associated mitochondrial dysfunction is mechanistically involved. Chlorogenic (CGA), a potent antioxidant and anti-inflammatory compound, protects against high-fat-diet-induced oxidative stress and mitochondrial dysfunction in . This study investigates whether CGA protects against FFA-induced hepatocyte lipotoxicity via the regulation of mitochondrial fission/fusion and elucidates its underlying mechanisms.AML12 cell, a non-transformed hepatocyte cell line, is treated with palmitate. Here, it is shown that CGA prevents palmitate-induced lipotoxicity by activation of SIRT1 regulated mitochondrial morphology. CGA treatment mitigates oxidative stress and mitochondrial dysfunction, as evidenced by a decrease in reactive oxygen species (ROS) production, and an increase in mitochondrial mass and mitochondrial membrane potential. CGA also significantly decreases Bax expression and thereby reduces mitochondria-mediated caspase-dependent apoptosis. Mechanistically, CGA attenuates ROS-induced mitochondrial fragmentation by inhibiting dynamin-related protein 1 (Drp1) and enhancing Mfn2 expression. In contrast, the inhibitory effects of CGA on the generation of mitochondrial ROS and Drp1 are blocked by siRNA knockdown of SIRT1.Collectively, these findings show that supplementation with CGA protects hepatocytes from FFA-induced lipotoxicity through activation of SIRT1, which reverses the oxidative stress and dysfunction of mitochondrial biogenesis directly.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic insulin resistance development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Oleic protects saturated mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis.

Aim This study aims to demonstrate the protective effects of monounsaturated oleic (OA) against saturated (PA) induced cellular lipotoxicity in hepatocytes and rats with non-alcoholic steatohepatitis (NASH).Human hepatoma cell line HepG2 cells and neonatal rat primary hepatocytes were treated with PA or/and OA for 24\u202fh. SD rats were fed with high fat diet (HFD) to induce NASH. From the 16th w, the HFD was full or half replaced by olive oil to observe the protective effects.In vitro, OA substantially alleviated PA induced cellular apoptosis, oxidative stress, ER stress, mitochondrial dysfunction, as well as inflammation in hepatocytes. In vivo, only olive oil supplementation had no detrimental effects, while HFD developed NASH in normal rats. Full replacement of HFD with olive oil had profoundly reversed NASH. Noteworthily, half replacement of HFD with olive oil (a mixed diet) has ameliorated NASH injury as well. It strikingly changed the hepatic histology from macrovesicular-steatosis into entire microvesicular-steatosis, and significantly reduced inflammation, ballooning and fibrosis.Our study has demonstrated in both hepatocytes and NASH rats that oleic acids had great potential to combat the saturated acids induced hepatic lipotoxicity. Only half replacement of HFD by monounsaturated acids rich diet still had significant therapeutic outcome in NASH rats. Redirecting the toxic saturated acids into triglyceride storage and reduction of cholesterol accumulation might be the possible explanation of OA driven protection in this scenario.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of acids in to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: fatty liver

Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated acids (PUFAs) and monounsaturated acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut microbiota, and hepatic metabolism.

Keyword: fatty liver

Serum lipids as an indicator for the alteration of function in patients with hepatitis B.

Hepatitis B virus (HBV) exerts an intense impact on host lipid metabolism. Hence the aim of present study is to determine metabolic derangement that occurred in subjects suffering from hepatitis B patients.The fasting blood samples were collected from hepatitis B patients (n\u2009=\u200950) attended in Taluka hospital TandoAdam, Sindh with age and gender matched controls (n\u2009=\u200950). Serum lipid profile and (FA) composition were analyzed by micro-lab and gas chromatography.The hepatitis B patients have significantly lower level (p\u2009<\u20090.01) of lipid profile including total cholesterol (TC), triacylglyceride (TAG), high density lipoprotein-C (HDL-C) very low density lipoprotein-cholesterol (VLDL-C), low density lipoprotein-cholesterol (LDL-C), and total lipid (TL) in comparison to controls, indicating hypolipidemia in patients. The result of total FA composition of HBV patients in comparison to controls reveal that myristic, , docosahexaenoic acids were significantly (p\u2009<\u20090.05) higher, while linoleic, eicosatrienoic, arachidonic, eicosapentaenoic acids were lower in HBV patients in comparison to controls. The elongase, ∆5 and ∆6-desaturase enzymes activities were found lower, while ∆9-desaturase activity was higher in hepatitis B patients as compared to controls, which indicates the impaired lipid metabolism.The serum saturated (SFA) and monounsaturated (MUFA) were increased while polyunsaturated (PUFA) was reduced in both total and free form in hepatitis B patients due to altered activities of enzyme desaturases with impaired PUFA metabolism and non-enzymatic oxidation.

Keyword: fatty liver

High α-tocopherol dosing increases lipid metabolism by changing redox state in damaged rat gastric mucosa and after ethanol treatment.

Regeneration of ethanol-injured rat gastric mucosa must undergo changes in major metabolic pathways to achieve DNA replication and cell proliferation. These events are highly dependent on glucose utilization and inhibited by vitamin E (VE) (α-tocopherol) administration. Therefore, the present study aimed at assessing lipid metabolism in the gastric mucosa and ethanol-induced gastric damage and the effect of α-tocopherol administration. For this, rates of β-oxidation and lipogenesis were tested in gastric mucosa samples. Through histological analysis, we found loss of the mucosa\'s superficial epithelium, which became gradually normalized during the recovery period. Proliferation of gastric mucosa occurred with augmented formation of β-oxidation by-products, diminished synthesis of triacylglycerols (TGs), as well as of phospholipids, and a reduced cytoplasmic NAD/NADH ratio, whereas the mitochondrial redox NAD/NADH ratio was much less affected. In addition, α-tocopherol increased utilization in the gastric mucosa, which was accompanied by the induction of \'mirror image\' effects on the cell redox state, reflected in an inhibited cell gastric mucosa proliferation by the vitamin administration. In conclusion, the present study shows, for the first time, the role of lipid metabolism in the adaptive cell gastric mucosa changes that drive proliferation after a chronic insult. Moreover, α-tocopherol increased gastric mucosa utilization of associated with energy production. These events could be associated with its antioxidant properties in co-ordination with regulation of genes and cell pathways, including changes in the cell NAD/NADH redox state.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fatty liver

Saturated combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic disease (NAFLD) is the most common chronic disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic inflammation, a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: fatty liver

Increased expression of sterol regulatory element binding protein‑2 alleviates autophagic dysfunction in NAFLD.

Sterol regulatory element binding protein‑2 (SREBP‑2) is an important transcription factor in lipid homeostasis. A previous study showed that SREBP‑2 also activated autophagic genes during cell‑sterol depletion. Alterations in autophagy are reported to be involved in the pathogenesis of nonalcoholic disease (NAFLD). However, whether the regulation of SREBP‑2 restores dysfunctional autophagy in hepatocytes during NAFLD remains to be elucidated. In the present study, a steatosis model was established with (PA) treatment at the indicated times and concentrations. Autophagosomes in hepatocytes were visualized by confocal microscopy after transfection with a tandem GFP‑mCherry‑LC3 construct. Autophagy‑associated protein levels were analyzed by western blot analysis. Loss‑ and gain‑of‑function studies were performed to examine the role of SREBP‑2 in the regulation of hepatocyte autophagy. It was demonstrated that PA induced autophagy and enhanced autophagic flux at the early stage, whereas prolonged treatment with PA resulted in dysfunction of autophagy in the PA‑induced steatotic hepatocytes. In addition, different cellular models presented with differing dysfunctional autophagy in response to overload. It was also confirmed that SREBP‑2 regulated autophagy‑related gene expression in hepatocytes, and it was shown that the overexpression of SREBP‑2 increased the expression of autophagy‑related genes, but did not affect the inhibition of the autophagic flux in lipid‑overloaded HL‑7702 cells. By contrast, increased SREBP‑2 partly restored the inhibited autophagic activity in lipid‑overloaded hepatoma HepG2 cells. Taken together, the present study demonstrated that autophagic function was impaired in lipid‑overloaded human hepatocytes, and the differential effect of PA on autophagy was associated with the duration of PA and the cell type. Under these conditions, the overexpression of SREBP‑2 alleviated the inhibited autophagic activity rather than the inhibition of autophagic flux. Consequently, the results indicated that restoration of autophagy dysfunction via the regulation of SREBP‑2 may be a potential therapeutic target for the treatment of NAFLD.

Keyword: fatty liver

Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic to nonalcoholic steatohepatitis.

Activation of inflammation is an important mechanism in the development of nonalcoholic steatohepatitis (NASH). This study aims to delineate how mitophagy affects NLRP3 inflammasome activation in hepatic lipotoxicity. Mice were fed a high fat/calorie diet (HFCD) for 24 weeks. Primary rat hepatocytes were treated with (PA) for various periods of time. Mitophagy was measured by protein levels of LC3II and P62. NLRP3, caspase-1, interleukin (IL)-18, and IL-1β at mRNA and protein levels were used as indicators of inflammasome activation. Along with steatotic progression in HFCD-fed mice, ratio of LC3II/β-actin was decreased concurrently with increased levels of P62, NLRP3, caspase-1, IL-1β, IL-18, and serum IL-1β levels in late-stage NASH. PA treatment resulted in mitochondrial oxidative stress and initiated mitophagy in primary hepatocytes. The addition of cyclosporine A did not change LC3II/Τοmm20 ratios; but P62 levels were increased after an extended duration of PA exposure, indicating a defect in autophagic activity. Along with impaired mitophagy, mRNA and protein levels of NLRP3, caspase-1, IL-18 and IL-1β were upregulated by PA treatment. Pretreatment with MCC950, N-acetyl cysteine or acetyl-L-carnitine reversed inflammasome activation and a pyroptotic cascade. Additionally, mitophagic flux was partially recovered as indicated by increases in LC3II/Tomm20 ratio, parkin, and PINK1 expression, and decreased P62 expression. The findings suggest that impaired mitophagy triggers hepatic NLRP3 inflammasome activation in a murine NASH model and primary hepatocytes. The new insights into inflammasome activation through mitophagy advance our understanding of how acids elicit lipotoxicity through oxidant stress and autophagy in mitochondria.

Keyword: fatty liver

Genomics of lipid-laden human hepatocyte cultures enables drug target screening for the treatment of non-alcoholic disease.

Non-alcoholic disease (NAFLD) is a major health burden in need for new medication. To identify potential drug targets a genomic study was performed in lipid-laden primary human hepatocyte (PHH) and human hepatoma cell cultures.PHH, HuH7 and HepG2 hepatoma cell cultures were treated with lipids and/or TNFα. Intracellular lipid load was quantified with the ORO assay. The Affymetrix HG-U133+ array system was employed to perform transcriptome analysis. The lipid droplet (LD) growth and fusion was determined by fluorescence microscopy. LD associated proteins were imaged by confocal immunofluorescence microscopy and confirmed by Western immunoblotting. Bioinformatics defined perturbed metabolic pathways.Whole genome expression profiling identified 227, 1031 and 571 significant regulated genes. Likewise, the combined lipid and TNFα treatment of PHH, HuH7 and HepG2 cell cultures revealed 154, 1238 and 278 differentially expressed genes. Although genomic responses differed among in-vitro systems, commonalities were ascertained by filtering the data for LD associated gene regulations. Among others the LD-growth and fusion associated cell death inducing DFFA like effector C (CIDEC), perilipins (PLIN2, PLIN3), the synaptosome-associated-protein 23 and the vesicle associated membrane protein 3 were strongly up-regulated. Likewise, the PPAR targets pyruvate-dehydrogenase-kinase-4 and angiopoietin-like-4 were up-regulated as was hypoxia-inducible lipid droplet-associated (HILPDA), flotilin and FGF21. Their inhibition ameliorates triglyceride and cholesterol accumulation. TNFα treatment elicited strong induction of the chemokine CXCL8, the kinases MAP3K8, MAP4K4 and negative regulators of cytokine signaling, i.e. SOCS2&SOCS3. Live cell imaging of DsRED calreticulin plasmid transfected HuH7 cells permitted an assessment of LD growth and fusion and confocal immunofluorescence microscopy evidenced induced LD-associated PLIN2, CIDEC, HIF1α, HILPDA, JAK1, PDK4 and ROCK2 expression. Notwithstanding, CPT1A protein was repressed to protect mitochondria from lipid overload. Pharmacological inhibition of the GTPase-dynamin and the transporter-2 reduced lipid uptake by 28.5 and 35%, respectively. Finally, a comparisons of in-vitro/NAFLD patient biopsy findings confirmed common gene regulations thus demonstrating clinical relevance.The genomics of fat-laden hepatocytes revealed LD-associated gene regulations and perturbed metabolic pathways. Immunofluorescence microscopy confirmed expression of coded proteins to provide a rationale for therapeutic intervention strategies. Collectively, the in-vitro system permits testing of drug candidates.

Keyword: fatty liver

Hepatocyte miR-33a mediates mitochondrial dysfunction and hepatosteatosis by suppressing NDUFA5.

Emerging evidence suggests that microRNAs (miRNAs) are essential for metabolic haemostasis of tissues. Among them, miR-33a is supposed to modulate the cholesterol export and oxidation, but whether miR-33a involves in the process of disease is unclear. To disclose the hypothesis, we utilized miR-33a mimic and antisense to explore their effects in primary hepatocytes or high-fat diet (HFD)-fed mice. Treatment with (PA) or HFD significantly increased the expression of miR-33a in hepatocytes or tissues. In primary hepatocytes, miR-33a mimic decreased mitochondrial function, including reduction of ATP production and oxygen consumption, whereas miR-33a inhibition protected PA-induced mitochondrial dysfunction. Interestingly, miR-33a selectively suppressed mitochondrial complex I activity and protein expression, but not other complexes. Through bioinformatics prediction, we found miR-33a directly targeted on the 3\'-UTR of NDUFA5. Dual-luciferase reporter analysis further confirmed the direct suppression of miR-33a on NDUFA5 expression. More importantly, administration of miR-33a antisense could effectively restore HFD-induced mitochondrial dysfunction through up-regulation of NDUFA5 levels. Mice treated with miR-33a antisense also exhibited improved function and structural disorders under obese status. Taken together, miR-33a was an important mediator of hepatocyte mitochondrial function, and the therapeutic benefits implied miR-33a antisense had the potential clinical application in combating the disease.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

Dihydromyricetin Ameliorates Nonalcoholic Disease by Improving Mitochondrial Respiratory Capacity and Redox Homeostasis Through Modulation of SIRT3 Signaling.

Our previous clinical trial indicated that the flavonoid dihydromyricetin (DHM) could improve hepatic steatosis in patients with nonalcoholic disease (NAFLD), altough the potential mechanisms of these effects remained elusive. Here, we investigated the hepatoprotective role of DHM on high-fat diet (HFD)-induced NAFLD. DHM supplementation could effectively ameliorate the development of NAFLD by inhibiting hepatic lipid accumulation both in HFD-fed wild-type mice and in -induced hepatocytes. We reveal for the first time that mitochondrial dysfunction characterized by ATP depletion and augmented oxidative stress could be reversed by DHM treatment. Moreover, DHM enhanced the mitochondrial respiratory capacity by increasing the expression and enzymatic activities of mitochondrial complexes and increased mitochondrial reactive oxygen species scavenging by restoring manganese superoxide dismutase (SOD2) activity. Interestingly, the benefits of DHM were abrogated in knockout (SIRT3KO) mice and in hepatocytes transfected with siRNA or treated with an SIRT3-specific inhibitor. We further showed that DHM could increase SIRT3 expression by activating the adenosine monophosphate-activated protein kinase (AMPK)-peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC1α)/estrogen-related receptor-α (ERRα) signaling pathway. Our work indicates that SIRT3 plays a critical role in the DHM-mediated beneficial effects that include ameliorating mitochondrial dysfunction and oxidative stress in a nutritional NAFLD model both and Our results suggest that DHM prevents NAFLD by improving mitochondrial respiratory capacity and redox homeostasis in hepatocytes through a SIRT3-dependent mechanism. These results could provide a foundation to identify new DHM-based preventive and therapeutic strategies for NAFLD.

Keyword: fatty liver

N-3 Polyunsaturated Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Diabetes Rats by Postponing Shortening of Hepatic Telomeres and Modulating Metabolism.

The long-term influence of gestational diabetes mellitus (GDM) on offspring and the effect of omega-3 polyunsaturated acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and inflammation in the of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the , in which numerous metabolites and metabolic pathways were altered when GDM offspring grew to old age. Many metabolites were related to diabetes risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious diabetes risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: fatty liver

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the of high-fat and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Keyword: fatty liver

miR-192-5p regulates lipid synthesis in non-alcoholic disease through SCD-1.

To evaluate the levels of miR-192-5p in non-alcoholic disease (NAFLD) models and demonstrate the role of miR-192-5p in lipid accumulation.Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet (HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic miR-192-5p and stearoyl-CoA desaturase 1 (SCD-1) levels were measured. MiR-192-5p mimic and inhibitor and SCD-1 siRNA were transfected into Huh7 cells exposed to (PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic miR-192-5p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection ( < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of miR-192-5p and SCD-1 protein levels, respectively ( < 0.01). Transfection with miR-192-5p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively ( < 0.01). Luciferase activity was suppressed and enhanced by miR-192-5p mimic and inhibitor, respectively, in wild-type SCD-1 ( < 0.01) but not in mutant SCD-1. MiR-192-5p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 siRNA transfection abrogated the lipid deposition aggravated by miR-192-5p inhibitor ( < 0.01).This study demonstrates that miR-192-5p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.

Keyword: fatty liver

Hepatocyte-Derived Lipotoxic Extracellular Vesicle Sphingosine 1-Phosphate Induces Macrophage Chemotaxis.

The pathophysiology of non-alcoholic steatohepatitis involves hepatocyte lipotoxicity due to excess saturated free acids and concomitant proinflammatory macrophage effector responses. These include the infiltration of macrophages into hepatic cords in response to incompletely understood stimuli. Stressed hepatocytes release an increased number of extracellular vesicles (EVs), which are known to participate in intercellular signaling and coordination of the behavior of immune cell populations via their cargo. We hypothesized that hepatocyte-derived lipotoxic EVs that are enriched in sphingosine 1-phosphate (S1P) are effectors of macrophage infiltration in the hepatic microenvironment. Lipotoxic EVs were isolated from palmitate treated immortalized mouse hepatocytes and characterized by nanoparticle tracking analysis. Lipotoxic EV sphingolipids were quantified using tandem mass spectrometry. Wildtype and S1P receptor knockout bone marrow-derived macrophages were exposed to lipotoxic EV gradients in a microfluidic gradient generator. Macrophage migration toward EV gradients was captured by time-lapse microscopy and analyzed to determine directional migration. Fluorescence-activated cell sorting along with quantitative PCR and immunohistochemistry were utilized to characterize the cell surface expression of S1P receptor on intrahepatic leukocytes and hepatic expression of S1P receptor, respectively. Palmitate treatment induced the release of EVs. These EVs were enriched in S1P. Palmitate-induced S1P enriched EVs were chemoattractive to macrophages. EV S1P enrichment depended on the activity of sphingosine kinases 1 and 2, such that, pharmacological inhibition of sphingosine kinases 1 and 2 resulted in a significant reduction in EV S1P cargo without affecting the number of EVs released. When exposed to EVs derived from cells treated with palmitate in the presence of a pharmacologic inhibitor of sphingosine kinases 1 and 2, macrophages displayed diminished chemotactic behavior. To determine receptor-ligand specificity, we tested the migration responses of macrophages genetically deleted in the S1P receptor toward lipotoxic EVs. S1P receptor knockout macrophages displayed a marked reduction in their chemotactic responses toward lipotoxic palmitate-induced EVs. Palmitate-induced lipotoxic EVs are enriched in S1P through sphingosine kinases 1 and 2. S1P-enriched EVs activate persistent and directional macrophage chemotaxis mediated by the S1P receptor, a potential signaling axis for macrophage infiltration during hepatic lipotoxicity, and a potential therapeutic target for non-alcoholic steatohepatitis.

Keyword: fatty liver

Macrophage-Specific Hypoxia-Inducible Factor-1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis.

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia-inducible factors (HIFs) have recently been identified as important regulators of immunity and inflammation. In nonalcoholic steatohepatitis (NASH), HIF-1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF-1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF-1α levels in hepatic macrophages in methionine-choline-deficient (MCD) diet-fed mice and in macrophages of patients with NASH compared with controls. The HIF-1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin-1, LC3-II, and p62 in both mouse and human macrophages. LysM HIF fl/fl mice, which have HIF-1α levels stabilized in macrophages, showed higher steatosis and inflammation compared with HIF fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated , (PA), both induces HIF-1α and impairs autophagic flux in macrophages. Using small interfering RNA-mediated knock-down and overexpression of HIF-1α in macrophages, we demonstrated that PA impairs autophagy via HIF-1α. We found that HIF-1α mediates NF-κB activation and MCP-1 production and that HIF-1α-mediated impairment of macrophage autophagy increases IL-1β production, contributing to MCD diet-induced NASH. Conclusion: impairs autophagy via HIF-1α activation in macrophages. HIF-1α and impaired autophagy are present in NASH in vivo in mouse macrophages and in human blood monocytes. We identified that HIF-1α activation and decreased autophagic flux stimulate inflammation in macrophages through upregulation of NF-κB activation. These results suggest that macrophage activation in NASH involves a complex interplay between HIF-1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet-induced NASH.© 2018 by the American Association for the Study of Diseases.

Keyword: fatty liver

Acetone fraction from Sechium edule (Jacq.) S.w. edible roots exhibits anti-endothelial dysfunction activity.

A recent ethnomedical survey on medicinal plants grown in Mexico revealed that Sechium edule (Jacq.) Sw. (Cucurbitaceae) is one of the most valued plant species to treat cardiovascular diseases, including hypertension. Fruits, young leaves, buds, stems, and tuberous roots of the plant are edible. Considering that endothelial dysfunction induced by Angiotensin II plays an important role in the pathogenesis of hypertension and is accompanied by a prooxidative condition, which in turn induces an inflammatory state, vascular remodeling, and tissue damage, and that S. edule has been reported to possess antioxidant, anti-inflammatory and antihypertensive activity, its capability to control endothelial dysfunction was also assessed.To assess in vivo the anti-endothelial dysfunction activity of the acetone fraction (rSe-ACE) of the hydroalcoholic extract from S. edule roots.Endothelial dysfunction was induced in female C57BL/6\u202fJ mice by a daily intraperitoneal injection of angiotensin II for 10 weeks. Either rSe-ACE or losartan (as a control) were co-administered with angiotensin II for the same period. Blood pressure was measured at weeks 0, 5, and 10. Kidney extracts were prepared to determine IL1β, IL4, IL6, IL10, IL17, IFNγ, TNFα, and TGFβ levels by ELISA, along with the prooxidative status as assessed by the activity of antioxidant enzymes. The expression of ICAM-1 was evaluated by immunohistochemistry in kidney histological sections. Kidney and hepatic damage, as well as vascular tissue remodeling, were studied.The rSe-ACE fraction administered at a dose of 10\u202fmg/kg was able to control hypertension, as well as the prooxidative and proinflammatory status in kidney as efficiently as losartan, returning mice to normotensive levels. Additionally, the fraction was more efficient than losartan to prevent and kidney damage. Phytochemical characterization identified cinnamic as a major compound, and linoleic, , and myristic acids as the most abundant non-polar components in the mixture, previously reported to aid in the control of hypertension, inflammation, and oxidative stress, three important components of endothelial dysfunction.this study demonstrated that rSe-ACE has anti-endothelial dysfunction activity in an experimental model and highlights the role of cinnamic and acids in the observed effects.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Acanthoic modulates lipogenesis in nonalcoholic disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic disease (NAFLD) associated with lipid accumulation by activating Farnesoid X receptor (FXR) and X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high-fat (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on lipid accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, index, and lipid droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated lipid accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating lipid accumulation and synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Advanced Fibrosis Is Independently Associated with and Insulin Levels in Patients with Non-Alcoholic Disease.

Changes in lipid metabolism occur during the development and progression non-alcoholic disease (NAFLD). However, the (FA) profile in red blood cells (RBC) from patients with fibrosis remains unexplored. Thus, the goal of this study was to evaluate the profile in RBC, dietary lipid intake and insulin resistance indicators in patients with NAFLD, according to the degree of hepatic fibrosis. Using elastography, patients were classified with ( = 52) and without ( = 37) advanced fibrosis. The profile in RBC was analyzed using gas chromatography and the lipid intake was evaluated through a 24-h dietary recall. Subjects with advanced fibrosis had higher levels of , stearic and oleic and total monounsaturated (MUFA) and insulin ( < 0.05), and lower levels of elongase very long chain acids protein-6 and the delta-5-desaturase enzymatic activity ( < 0.05). These results suggest a lack of regulation of enzymes related to FA metabolism in patients with advanced fibrosis.

Keyword: fatty liver

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fatty liver

Euodia daniellii Hemsl. (Bee-Bee Tree) Oil Attenuates Palmitate-Induced Lipid Accumulation and Apoptosis in Hepatocytes.

Hepatic lipid accumulation and apoptosis is elevated in patients with non-alcoholic steatohepatitis and is closely associated with severity. Saturated palmitate stimulates lipid accumulation and apoptosis in hepatocytes. In the present study, we examined bee-bee tree oil (BO)-mediated protective effects on palmitate-induced lipid accumulation and apoptosis in mouse primary hepatocytes. Cells were cultured in a control media or the same media containing 150 or 300 µmol/L of albumin-bound palmitate for 24 h. BO concentrations used were 0, 0.1, 0.2, or 0.5%. Palmitate induced lipid accumulation and mRNA expression of lipogenic genes such as SREBP1c and SCD1. However, BO prevented these changes. Furthermore, palmitate stimulated caspase-3 activity and decreased cell viability in the absence of BO. BO reduced palmitate-induced activation of caspase-3 and cell death in a dose-dependent manner. AMP-activated protein kinase inhibitors abolished the effects of BO. Furthermore, BO suppressed palmitate-induced c-Jun N-terminal kinase (JNK) phosphorylation through the 5' adenosine monophosphate-activated protein kinase (AMPK)-dependent pathway. In conclusion, BO attenuated palmitate-induced hepatic steatosis and apoptosis through AMPK-mediated suppression of JNK signaling. These data suggest that BO is an important determinant of saturated -induced lipid accumulation and apoptosis, and may be an effective therapeutic strategy for treatment of obesity-mediated diseases.© 2018 S. Karger AG, Basel.

Keyword: fatty liver

d- chiro-Inositol Ameliorates High Fat Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high fat diet and saturated -treated hepatocytes. DCI attenuated free uptake by the via lipid trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces lipid deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: fatty liver

acids uptake and oxidation are increased in the of rats with adjuvant-induced arthritis.

Severe rheumatoid cachexia is associated with pronounced loss of muscle and fat mass in patients with advanced rheumatoid arthritis. This condition is associated with dyslipidemia and predisposition to cardiovascular diseases. Circulating levels of triglycerides (TG) and free acids (FFA) have not yet been consistently defined in severe arthritis. Similarly, the metabolism of these lipids in the arthritic has not yet been clarified. Aiming at filling these gaps this study presents a characterization of the circulating lipid profile and of the acids uptake and metabolism in perfused livers of rats with adjuvant-induced arthritis. The levels of TG and total cholesterol were reduced in both serum (10-20%) and (20-35%) of arthritic rats. The levels of circulating FFA were 40% higher in arthritic rats, possibly in consequence of cytokine-induced adipose tissue lipolysis. Hepatic uptake and oxidation of and oleic acids was higher in arthritic livers. The phenomenon results possibly from a more oxidized state of the arthritic . Indeed, NADPH/NADP and NADH/NAD ratios were 30% lower in arthritic livers, which additionally presented higher activities of the citric cycle driven by both endogenous and exogenous FFA. The lower levels of circulating and hepatic TG possibly are caused by an increased oxidation associated to a reduced synthesis of acids in arthritic livers. These results reveal that the lipid hepatic metabolism in arthritic rats presents a strong catabolic tendency, a condition that should contribute to the marked cachexia described for arthritic rats and possibly for the severe rheumatoid arthritis.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and . PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: fatty liver

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid Metabolism, Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free (FFA)- and oleic -and high-fat diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice were measured, and the expression levels of genes associated with lipogenesis, oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of lipid metabolism, inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: fatty liver

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and obesity in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

Keyword: fatty liver

ester of tetrahydrocannabinol (THC) and diester of 11-hydroxy-THC - Unsuccessful search for additional THC metabolites in human body fluids and tissues.

conjugates of hydroxy-metabolites of tetrahydrocannabinol (THC) or cannabinol have already been reported as metabolites in rats. In the herein presented investigation, esters of THC and its primary metabolite 11-hydroxy-delta9-tetrahydrocannabinol (11-OH-THC) were synthesized using esterification with chloride. Structural elucidation of the products was conducted using nuclear magnetic resonance spectroscopy (NMR) and liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC-QToF-MS). For the confirmation of a previous cannabis use, body fluids (femoral blood, heart blood, urine, bile) of 27 death cases (all with known cannabis use), including adipose tissue homogenates of six of these cases as well as eleven plasma samples (probably all with regular cannabis use, confirmed by a high 11-nor-9-carboxy-delta9-tetrahydrocannabinol (THC-COOH) concentration (except one sample, >200ng/mL), were tested for THC and its main metabolites 11-OH-THC and THC-COOH using gas chromatography coupled to mass spectrometry (GC-MS). These samples as well as further tissue homogenates of autopsy cases (, kidney, brain) were additionally tested for the presence of THC ester or 11-OH-THC diester by means of a liquid chromatographic triple quadrupole mass spectrometric (LC-QQQ-MS) method, in order to evaluate a possible presence of these conjugates in humans. In none of the analyzed samples (in total 196 specimens; plasma (N=11), femoral blood (N=23), heart blood (N=25), urine (N=23), bile (N=27), (N=27), kidney (N=27), brain (N=27), adipose tissue (N=6)), esters of THC or 11-OH-THC could be proven. Even if the existence of these esters in human samples cannot be ruled out definitely, suitability as cannabis consumption markers does not seem likely based on our findings.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Treatment of cigarette smoke extract and condensate differentially potentiates -induced lipotoxicity and steatohepatitis in vitro.

Accumulative evidence showed that cigarette smoke (CS) detrimentally affects the pathogenesis of nonalcoholic steatohepatitis (NASH). The purpose of this study was to evaluate the effects of CS extract (CSE) or total particulate matter (TPM) on the in vitro steatohepatitis model using mouse primary hepatocytes treated with (PA) or PA plus LPS. Increased hepatocellular damage was observed in PA-treated hepatocytes with TPM or CSE treatment, but increased triglyceride level was only observed in PA plus LPS-treated hepatocytes with a high concentration of TPM. Also, expression levels of steatohepatitis-related genes such as TNF-α, NOS 2, and SREBP-1c were significantly increased after treatment of TPM. To further demonstrate the role of Kupffer cells (KCs) after CS extracts treatment, trans-well co-culture system of hepatocytes and KCs was utilized. The levels of inflammatory cytokines and the ratios of Bax/Bcl-2 (apoptosis-related genes) were markedly increased in co-cultured hepatocytes after TPM or CSE treatment. Interestingly, KCs activation was augmented in KCs upon treatment with CSE or TPM. Overall, our findings indicate that in vitro treatment with CSE or TPM differentially contributes to the severity of steatohepatitis by modulating steatohepatitis-related lipotoxicity and inflammation, which might be caused by KCs activation with subsequent induction of hepatocytes apoptosis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Cathepsin B inhibition ameliorates the non-alcoholic steatohepatitis through suppressing caspase-1 activation.

Non-alcoholic disease (NAFLD) has emerged as the most common chronic disease. NLRP3 inflammasome activation has been widely studied in the pathogenesis of NAFLD. Cathepsin B (CTSB) is a ubiquitous cysteine cathepsin, and the role of CTSB in the progression and development of NAFLD has received extensive concern. However, the exact roles of CTSB in the NAFLD development and NLRP3 inflammasome activation are yet to be evaluated. In the present study, we used methionine choline-deficient (MCD) diet to establish mice NASH model. CTSB inhibitor (CA-074) was used to suppress the expression of CSTB. Expressions of CTSB and caspase-1 were evaluated by immunohistochemical staining. Serum IL-1β and IL-18 levels were also determined. was used to stimulate Kupffer cells (KCs), and protein expressions of CTSB, NLRP3, ASC (apoptosis-associated speck-like protein containing CARD), and caspase-1 in KCs were detected. The levels of IL-1β and IL-18 in the supernatant of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). Our results showed that CTSB inhibition improved the function and reduced hepatic inflammation and ballooning, and the levels of pro-inflammatory cytokines IL-1β and IL-18 were decreased. The expressions of CTSB and caspase-1 in tissues were increased in the NASH group. In in vitro experiments, PA stimulation could increase the expressions of CTSB and NLRP3 inflammasome in KCs, and CTSB inhibition downregulated the expression of NLRP3 inflammasome in KCs, when challenged by PA. Moreover, CTSB inhibition effectively suppressed the expression and activity of caspase-1 and subsequently secretions of IL-1β and IL-18. Collectively, these results suggest that CTSB inhibition limits NLRP3 inflammasome-dependent NASH formation through regulating the expression and activity of caspase-1, thus providing a novel anti-inflammatory signal pathway for the therapy of NAFLD.

Keyword: fatty liver

Unsaturated Acids Increase the Expression of Hepassocin through a Signal Transducer and Activator of Transcription 3-Dependent Pathway in HepG2 Cells.

Hepassocin (HPS) is a hepatokine that regulates hepatocyte proliferation. It is known that HPS plays an important role in the development of nonalcoholic diseases (NAFLD). acids, such as oleic (OLA), exhibit the ability to activate the signal transducer and activator of transcription-3 (STAT3), and the binding site of STAT3 is found in the promoter region of HPS. However, the regulation of HPS by acids is still obscure. To clarify the regulation of HPS, we detected the expression of HPS by western blots. In addition, a hepatic steatosis cell culture model was established by treatment of different acids, including linoleic (LNA), oleic , , and stearic . The intracellular lipid accumulation was confirmed by oil red O staining. Blocking of STAT3 activity was achieved by the pretreatment of the STAT3 inhibitor, stattic. We found that activation of STAT3 by interleukin-6 (IL-6) was mediated in the regulation of HPS expression. Treatment of unsaturated acids significantly induced intracellular lipid accumulation in HepG2 cells. Moreover, the expressions of HPS were increased in unsaturated -treated HepG2 cells, as compared with saturated -treated groups. Also, the expression of HPS induced by OLA was blocked by the inhibition of STAT3 activity. Furthermore, we found that deletion of HPS by small interfering ribonucleic transfection decreased the protective effect of OLA on cell viability. Taken together, we provided evidence that STAT3 plays an important role in the regulation of OLA-induced HPS expression and the increased HPS may further participate in the development of NAFLD. In addition, the increase of HPS might be involved in the protective effect of OLA on cell viability.© 2018 AOCS.

Keyword: fatty liver

Degradation of splicing factor SRSF3 contributes to progressive liver disease.

Serine rich splicing factor 3 (SRSF3) plays a critical role in liver function and its loss promotes chronic liver damage and regeneration. As a consequence, genetic deletion of SRSF3 in hepatocytes caused progressive liver disease and ultimately led to hepatocellular carcinoma. Here we show that SRSF3 is decreased in human liver samples with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or cirrhosis that was associated with alterations in RNA splicing of known SRSF3 target genes. Hepatic SRSF3 expression was similarly decreased and RNA splicing dysregulated in mouse models of NAFLD and NASH. We showed that -induced oxidative stress caused conjugation of the ubiquitin like NEDD8 protein to SRSF3 and proteasome mediated degradation. SRSF3 was selectively neddylated at lysine11 and mutation of this residue (SRSF3-K11R) was sufficient to prevent both SRSF3 degradation and alterations in RNA splicing. Finally prevention of SRSF3 degradation in vivo partially protected mice from hepatic steatosis, fibrosis and inflammation. These results highlight a neddylation-dependent mechanism regulating gene expression in the liver that is disrupted in early metabolic liver disease and may contribute to the progression to NASH, cirrhosis and ultimately hepatocellular carcinoma.

Keyword: fatty liver

Dihydrosterculic from cottonseed oil suppresses desaturase activity and improves metabolomic profiles of high-fat-fed mice.

Polyunsaturated (PUFA)-rich diets are thought to provide beneficial effects toward metabolic health in part through their bioactive properties. We hypothesized that increasing PUFA intake in mice would increase peroxisome proliferator activated receptor delta (PPARδ) expression and activity, and we sought to examine the effect of different PUFA-enriched oils on muscle PPARδ expression. One of the oils we tested was cottonseed oil (CSO) which is primarily linoleic (53%) and (24%). Mice fed a CSO-enriched diet (50% energy from fat) displayed no change in muscle PPARδ expression; however, in the , it was consistently elevated along with its transcriptional coactivator Pgc-1. Male mice were fed chow or CSO-, saturated fat (SFA)-, or linoleic (18:2)-enriched diets that were matched for macronutrient content for 4 weeks. There were no differences in food intake, body weight, fasting glucose, glucose tolerance, or energy expenditure between chow- and CSO-fed mice, whereas SFA-fed mice had increased fat mass and 18:2-fed mice were less glucose tolerant. Metabolomic analyses revealed that the livers of CSO-fed mice closely matched those of chow-fed but significantly differed from SFA- and 18:2-enriched groups. composition of the diets and livers revealed an impairment in desaturase activity and the presence of dihydrosterculic (DHSA) in the CSO-fed mice. The effect of DHSA on PPARδ and stearoyl-CoA desaturase-1 expression mimicked that of the CSO-fed mice. Taken together, these data suggest that DHSA from CSO may be an effective means to increase PPARδ expression with concomitant suppression of stearoyl-CoA desaturase-1 activity.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Water Extract of L. Ameliorates Non-Alcoholic Disease.

Our aim was to investigate whether hot water extract (CLW) of L. could prevent non-alcoholic disease (NAFLD). HepG2 cells were treated with free (FFA) mixture (oleic : , 2:1) for 24 h to stimulate in vitro . In addition, C57BL/6 mice were fed 60 kcal% high-fat (HF) diet for eight weeks to induce in vivo. Intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) productions were increased by FFA and HF-diet, but supplementation with CLW significantly decreased these levels. CLW treatment ameliorated antioxidant activities that were suppressed by exposure to the FFA and HF-diet. Cluster of differentiation 36 (CD36) and transport proteins (FATP2 and FATP5) were increased in HF-diet groups, while CLW suppressed their expression levels. Moreover, sterol regulatory element-binding protein-1c (SREBP-1c), acetyl-coenzyme A carboxylase (ACC), and synthase (FAS) expression levels were down-regulated in the CLW groups compared to HF-diet groups. On the other hand, 5\' adenosine monophosphate-activated protein kinase (AMPK), Peroxisome proliferator-activated receptor alpha (PPAR-α), and carnitine palmitoyltransferase 1 (CPT-1) expressions were up-regulated in the CLW groups. HF-diet fed mice showed high hepatic triglycerides (TG) content compared to the normal diet mice. However, the administration of CLW restored the hepatic TG level, indicating an inhibitory effect against lipid accumulation by CLW. These results suggest that CLW could be a potentially useful agent for the prevention of NAFLD through modulating uptake.

Keyword: fatty liver

Gut microbiota-mediated generation of saturated acids elicits inflammation in the in murine high-fat diet-induced steatohepatitis.

The gut microbiota plays crucial roles in the development of non-alcoholic steatohepatitis (NASH). However, the precise mechanisms by which alterations of the gut microbiota and its metabolism contributing to the pathogenesis of NASH are not yet fully elucidated.Mice were fed with a recently reported new class of high-fat diet (HFD), steatohepatitis-inducing HFD (STHD)-01 for 9\xa0weeks. The composition of the gut microbiota was analyzed by T-RFLP. Luminal metabolome was analyzed using capillary electrophoresis and liquid chromatography time-of-flight mass spectrometry (CE- and LC-TOFMS).Mice fed the STHD-01 developed NASH-like pathology within a short period. Treatment with antibiotics prevented the development of NASH by STHD-01. The composition of the gut microbiota and its metabolic activities were markedly perturbed in the STHD-01-fed mice, and antibiotic administration normalized these changes. We identified that long-chain saturated and n-6 metabolic pathways were significantly altered by STHD-01. Of note, the changes in gut lipidome caused by STHD-01 were mediated by gut microbiota, as the depletion of the gut microbiota could reverse the perturbation of these metabolic pathways. A saturated long-chain , , which accumulated in the STHD-01 group, activated macrophages and promoted TNF-α expression.Lipid metabolism by the gut microbiota, particularly the saturation of acids, affects fat accumulation in the and subsequent inflammation in NASH.

Keyword: fatty liver

Evidence for an alternative desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid metabolism that enables them to synthesize, elongate and desaturate acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target metabolism and, in particular, desaturation. This suggests that many cancer cells contain an unexplored plasticity in their metabolism. Here we show that some cancer cells can exploit an alternative desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human and lung carcinomas that desaturate palmitate to the unusual sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: fatty liver

Inhibition of MD2-dependent inflammation attenuates the progression of non-alcoholic disease.

Non-alcoholic disease (NAFLD) can progress to the more serious non-alcoholic steatohepatitis (NASH), characterized by inflammatory injury and fibrosis. The pathogenic basis of NAFLD progressing to NASH is currently unknown, but growing evidence suggests MD2 (myeloid differentiation factor 2), an accessory protein of TLR4, is an important signalling component contributing to this disease. We evaluated the effectiveness of the specific MD2 inhibitor, L6H21, in reducing inflammatory injury in a relevant high-fat diet (HFD) mouse model of NASH and in the (PA)-stimulated human cell line (HepG2). For study, genetic knockout (MD2 ) mice were fed a HFD or control diet for 24 weeks, or wild-type mice placed on a similar diet regimen and treated with L6H21 for the last 8 or 16 weeks. Results indicated that MD2 inhibition with L6H21 was as effective as MD2 knockout in preventing the HFD-induced hepatic lipid accumulation, pro-fibrotic changes and expression of pro-inflammatory molecules. Direct challenge of HepG2 with PA (200 μM) increased MD2-TLR4 complex formation and expression of pro-inflammatory and pro-fibrotic genes and L6H21 pre-treatment prevented these PA-induced responses. Interestingly, MD2 knockout or L6H21 increased expression of the anti-inflammatory molecule, PPARγ, in tissue and the cell line. Our results provide further evidence for the critical role of MD2 in the development of NASH and conclude that MD2 could be a potential therapeutic target for NAFLD/NASH treatment. Moreover, the small molecule MD2 inhibitor, L6H21, was an effective and selective investigative agent for future mechanistic studies of MD2.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

Liraglutide protects non-alcoholic disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high-fat diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease degeneration of the . The purpose of this study is to explore whether liraglutide improves high-fat diet-induced non-alcoholic disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the . After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the , /body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high-fat diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high-fat diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced obesity.

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti-obesity drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased body and weights, insulin, leptin, triglyceride, cholesterol and free plasma levels in a mouse model of diet-induced obesity. Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in energy expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in obesity and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

Keyword: fatty liver

Trimethylamine N-Oxide Aggravates Steatosis through Modulation of Bile Metabolism and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic disease (NAFLD) and could influence bile (BA) metabolism. However, whether TMAO aggravates steatosis by modulating BA metabolism and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA metabolism. Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

Natural Abundance Carbon Isotopic Analysis Indicates the Equal Contribution of Local Synthesis and Plasma Uptake to Palmitate Levels in the Mouse Brain.

Saturated acids are the most abundant acids in the brain, however, there has been some debate regarding the ability of intact dietary saturated acids to be incorporated into the brain. In the present study, we use compound specific isotope analysis to measure the natural abundance carbon isotopic signature of brain, , and blood (PAM) and compare it to the dietary PAM and sugar isotopic signatures to calculate the relative contribution of both the incorporation of intact and endogenously synthesized PAM to these pools. Mice were equilibrated to the study diet, and extracted acids were analyzed with gas chromatography isotope ratio mass spectrometry to determine the carbon isotopic signature of PAM (δ C ). , serum total, and serum unesterified δ C ranged between -20.6 and -21.1 mUr and were approximately 8.5 mUr more enriched in C when compared to the dietary PAM signature. Brain δ C was found to be more enriched than or blood pools (-16.7 ± 0.2 mUr, mean ± SD). Two end-member-mixed modeling using the carbon isotopic signature of dietary PAM and dietary sugars determined the contribution of synthesis to the total tissue PAM pool to range between 44% and 48%. This suggests that endogenous synthesis and dietary PAM are near equal contributors to brain, , and blood PAM pools. In conclusion, our data provide evidence that brain PAM levels are maintained by both local endogenous synthesis and through the uptake of intact PAM from the blood.© 2018 AOCS.

Keyword: fatty liver

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: fatty liver

Procyanidin B2 ameliorates free acids-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state.

Procyanidin B2, a naturally occurring phenolic compound, has been reported to exert multiple beneficial functions. However, the effect of procyanidin B2 on free acids (FFAs)-induced hepatic steatosis remains obscure. The present study is therefore aimed to elucidate the protective effect of procyanidin B2 against hepatic steatosis and its underlying mechanism. Herein, we reported that procyanidin B2 attenuated FFAs-induced lipid accumulation and its associated oxidative stress by scavenging excessive ROS and superoxide anion radicals, blocking loss of mitochondrial membrane potential, restoring glutathione content, and increasing activity of antioxidant enzymes (GPx, SOD and CAT) in hepatocytes. Procyanidin B2 mechanistically promoted lipid degradation via modulation of transcription factor EB (TFEB), a master regulator of lysosomal pathway. Molecular docking analysis indicated a possible ligand-binding position of procyanidin B2 with TFEB. In addition, administration of procyanidin B2 resulted in a significant reduction of hepatic fat accumulation in high-fat diet (HFD)-induced obese mice, and also ameliorated HFD-induced metabolic abnormalities, including hyperlipidemia and hyperglycemia. It was confirmed that procyanidin B2 prevented HFD-induced hepatic fat accumulation through down-regulating lipogenesis-related gene expressions (PPARγ, C/EBPα and SREBP-1c), inhibiting pro-inflammatory cytokines production (IL-6 and TNF-α) and increasing antioxidant enzymes activity (GPx, SOD and CAT). Moreover, hepatic acids analysis indicated that procyanidin B2 caused a significant increase in the levels of , oleic and linoleic . Intriguingly, procyanidin B2 restored the decreased nuclear TFEB expression in HFD-induced steatosis and up-regulated its target genes involved in lysosomal pathway (Lamp1, Mcoln, Uvrag), which suggested a previously unrecognized mechanism of procyanidin B2 on ameliorating HFD-induced hepatic steatosis. Taken together, our results demonstrated that procyanidin B2 attenuated FFAs-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state, which had important implications that modulation of TFEB might be a potential therapeutic strategy for hepatic steatosis and procyanidin B2 could represent a promising novel agent in the prevention and treatment of non-alcoholic disease (NAFLD).Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 diabetes mellitus (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high fat diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and lipid homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and , gastrocnemius and brown fat of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: fatty liver

[Glucagon-like peptide-1 regulates lipid metabolism in hepatocytes through Foxo1/3].

Glucagon-like peptide-1 (GLP-1) has been reported to be effective in the treatment of nonalcoholic disease (NAFLD). However, the molecular mechanism of GLP-1 on NAFLD is remained unclear. The present study was to detect whether the effect of GLP-1 on triglyceride (TG) content in hepatocytes is dependent on Foxos. HepG2 cells were treated with /oleic for 24 h. The knockdown of Foxo1, Foxo3 was conducted through small interfering RNA (siRNA). Real time PCT (RT-PCR) was used to detect the changes of the SREBP1c and Aco genes in HepG2 cells after Foxo1/3 knockdown. As expected, /oleic increased TG concentration in HepG2 cells [(12.65 ± 1.32) μg/mg vs. (4.32±0.54) μg/mg, 0.05]. Addition of GLP-1 dose (10, 50, 100nmol/L) dependently lowered the TG content and reached plateau at 100 nmol/L of GLP-1 [TG(8.38±1.47) μg/mg]. The GLP-1 effect on TG remained after knocking down either Foxo1 [(9.09±1.34)μg/mg] or Foxo3 [(8.90±1.60) μg/mg] alone, but not when knocking down Foxo1 and Foxo3 (Foxo1/3) together [(14.66±1.77)μg/mg]. Moreover, knocking down Foxo1/3 also abolished GLP-1 effect on SREBP1c and Aco expression. GLP-1 can inhibit the synthesis of TG in hepatocytes depending on Foxo1 and Foxo3. Further studies are needed to explore the specific mechanisms.

Keyword: fatty liver

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting lipogenesis and inflammation.

Non-alcoholic disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited lipid accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased lipid accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fatty liver

Resveratrol Maintains Lipid Metabolism Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver lipid metabolism disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver lipid metabolism disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid metabolic disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive oxygen species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular lipid metabolic imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of lipid metabolic disorders relevant to the circadian clock.

Keyword: fatty liver

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic disease (NAFLD) is the most common chronic disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy metabolism, but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic . The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: fatty liver

Serine prevented high-fat diet-induced oxidative stress by activating AMPK and epigenetically modulating the expression of glutathione synthesis-related genes.

Serine deficiency has been observed in patients with nonalcoholic disease (NAFLD). Whether serine supplementation has any beneficial effects on the prevention of NAFLD remains unknown. The present study was conducted to investigate the effects of serine supplementation on hepatic oxidative stress and steatosis and its related mechanisms. Forty male C57BL/6J mice (9week-old) were randomly assigned into four groups (n=10) and fed: i) a low-fat diet; ii) a low-fat diet supplemented with 1% (wt:vol) serine; iii) a high-fat (HF) diet; and iv) a HF diet supplemented with 1% serine, respectively. (PA)-treated primary hepatocytes separated from adult mice were also used to study the effects of serine on oxidative stress. The results showed that serine supplementation increased glucose tolerance and insulin sensitivity, and protected mice from hepatic lipid accumulation, but did not significantly decreased HF diet-induced weight gain. In addition, serine supplementation protected glutathione (GSH) antioxidant system and prevented hypermethylation in the promoters of glutathione synthesis-related genes, while decreasing reactive oxygen species (ROS) in mice fed a HF diet. Moreover, we found that serine supplementation increased phosphorylation and S-glutathionylation of AMP-activated protein kinase α subunit (AMPKα), and decreased ROS, malondialdehyde and triglyceride contents in PA-treated primary hepatocytes. However, while AMPK activity or GSH synthesis was inhibited, the abovementioned effects of serine on PA-treated primary hepatocytes were not observed. Our results suggest that serine supplementation could prevent HF diet-induced oxidative stress and steatosis by epigenetically modulating the expression of glutathione synthesis-related genes and through AMPK activation.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Acetyl-CoA from inflammation-induced acids oxidation promotes hepatic malate-aspartate shuttle activity and glycolysis.

Hepatic metabolic syndrome is associated with inflammation, as inflammation stimulates the reprogramming of nutrient metabolism and hepatic mitochondria-generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and acids, in the condition of inflammation is still unclear. Here, we used an acute inflammation model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic glycolysis and oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain glycolysis. With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by oxidation promotes MAS activity and glycolysis via nonenzymatic acetylation during the inflammatory stress response.

Keyword: fatty liver

plus Synergistically Ameliorates Nonalcoholic Steatohepatitis in HepG2 Cells.

The combination of and radix is frequently prescribed for diseases in TKM. However, the synergic effects of the two herbs on nonalcoholic steatohepatitis (NASH) have not yet been studied. Therefore, we investigated the anti-NASH effects of the water extract of (AI), radix (CL), and combination of the two herbs (ACE). Hepatic steatosis and NASH were induced in HepG2 cells by treatment with (PA, for 6\u2009h) with/without pretreatment of ACE (25 or 50\u2009g/mL), AI (50 or 100\u2009g/mL), CL (50 or 100\u2009g/mL), curcumin (5\u2009g/mL), or scopoletin (5\u2009g/mL). The PA treatment (200\u2009M) drastically altered intracellular triglyceride levels, total cholesterol, and expression levels of genes related to lipid metabolism (CD36, SREBP1c, PPAR-, and PPAR-), whereas pretreatment with ACE significantly attenuated these alterations. ACE also protected HepG2 cells from PA- (300\u2009M-) induced endoplasmic reticulum (ER) stress and apoptosis and attenuated the related key molecules including GRP78, eIF2, and CHOP, respectively. In conclusion, we found synergic effects of and on NASH, supporting the clinical potential for disorders. In addition, modulation of ER stress-relative molecules would be involved in its underlying mechanism.

Keyword: fatty liver

Serum C16:1n7/C16:0 ratio as a diagnostic marker for non-alcoholic steatohepatitis.

Accurate diagnosis of non-alcoholic steatohepatitis (NASH) from non-alcoholic disease (NAFLD) is clinically important. Therefore, there is a need for easier ways of diagnosing NASH. In this study, we investigated the serum composition and evaluated the possibility of using the serum composition as a diagnostic marker of NASH.The subjects were 78 NAFLD patients (non-alcoholic [NAFL]: 30, NASH: 48) and 24 healthy individuals. acids extracted from the tissue and serum were identified and quantified by gas chromatography. In addition, we evaluated the relationship between serum and tissue composition, patient background, and histology. The diagnostic performance of NASH was evaluated by calculating the area under the receiver operating characteristic (AUROC).The results of the analysis showed the C16:1n7/C16:0 ratio to have the strongest correlation between serum and tissue (r\xa0=\xa00.865, P\xa0<\xa00.0001). The serum C16:1n7/C16:0 ratio in the NASH group was higher compared with that in the NAFL group (P\xa0=\xa00.0007). Evaluation of the association of the serum C16:1n7/C16:0 ratio with histology revealed significant correlation with lobular inflammation score, ballooning score, and fibrosis score. The AUROC for predicting NASH in all NAFLD patients was 0.7097. The AUROC was nearly equivalent even when the study subjects were restricted to patients with a fibrosis score\xa0≤\xa02 only (AUROC 0.6917).Measuring the serum C16:1n7/C16:0 ratio may be an effective non-invasive method for diagnosing NASH, particularly in its early stages.© 2019 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular Inflammation in HepG2 Cells.

acids induced hepatic inflammation plays an important role in nonalcoholic disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti-inflammation in various human organs. However, the anti-inflammation property of HS in the is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free acids-induced inflammation. The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fatty liver

[Inhibition of chitin oligosaccharide on dyslipidemia and the potential molecular mechanism exploration].

The inhibitory effect of NACOS on dyslipidemia and potential molecular mechanisms by in vitro and in vivo experiments were investigated. For in vitro study, four experimental groups were designed by using HepG2 cells, including the control group, (PA) treatment alone group, NACOS treatment alone group and NACOS + PA treatment group. For in vivo study, male C57BL/6 mice were divided into four groups (n=5) at random including the normal control group (NCD), high fat diet (HFD) group, NACOS treatment alone group, NACOS+HFD group, which were treated for 20 weeks. The used methods in this study were as follows: the observation of lipid droplet deposition in HepG2 cells by oil red O staining, the detection of mRNA levels of lipid metabolism-related regulators and inflammatory cytokine by RT-PCR method, the monitoring of MAPKs and PI3K/Akt pathway activation by Western blotting method. The in vitro study shows that, NACOS had no toxicity on the viability of HepG2 cells at 25-100 μg/mL and significantly reduced the deposition of lipid droplet. Also, based on both in vitro and in vivo investigation, NACOS evidently down-regulated the expression of lipid metabolism-related regulators (PGC1α, Cox5b, Mcad) and inflammatory cytokine (IL-1β) at mRNA level (P<0.05 or 0.01), and suppressed the activation of p38, ERK1/2 and Akt in HepG2 cells and lever tissues from HFD-fed mice (P<0.05 or 0.01). Based on the above, NACOS may inhibit the oxidation of mitochondrial and the lipid biosynthesis, block the inflammatory responses and prevent the HepG2 cells and C57BL/6 mice from lipidemia.

Keyword: fatty liver

Human relaxin-2 attenuates hepatic steatosis and fibrosis in mice with non-alcoholic disease.

Human relaxin-2 reduces hepatic fibrosis in mice. However, the effects of relaxin-2 on hepatic steatosis and fibrosis in animals with non-alcoholic disease (NAFLD) remain to be elucidated. C57BL/6 mice fed a high-fat diet (HFD) or methionine-choline-deficient (MCD) diet were randomly assigned to receive recombinant human relaxin-2 (25 or 75\u2009μg/kg/day) or vehicle for 4 weeks. In HFD-fed mice, relaxin-2 decreased systemic insulin resistance and reduced body weight, epididymal fat mass and serum leptin and insulin concentrations. In livers of HFD-fed mice, relaxin-2 attenuated steatosis and increased phosphorylation of insulin receptor substrate-1, Akt and endothelial nitric oxide synthase (eNOS), and activated genes that regulate oxidation and suppressed acetyl-CoA carboxylase. Relaxin-2 had no direct anti-steatotic effect on primary mouse hepatocytes, but S-nitroso-N-acetylpenicillamine attenuated -induced steatosis and activated genes regulating oxidation in hepatocytes. In mice fed an MCD diet, relaxin-2 attenuated steatosis, inflammation and fibrosis. Relaxin-2 increased eNOS and Akt phosphorylation and transcript levels of cytochrome P450-4a10 and decreased acetyl-CoA carboxylase in MCD-fed mouse livers. Moreover, expression levels of Kupffer cell activation, hepatic stellate cell activation and hepatocyte apoptosis were decreased in MCD diet-fed mice receiving relaxin-2. In conclusion, relaxin-2 reduces hepatic steatosis by activating intrahepatic eNOS in HFD-fed mice and further attenuates fibrosis in MCD diet-fed mice. Therefore, human relaxin-2 is a potential therapeutic treatment for NAFLD.

Keyword: fatty liver

Increased serum concentration of ceramides in obese children with nonalcoholic disease.

Hepatic lipid accumulation is closely related to the development of insulin resistance, which is regarded as one of the most significant risk factors of nonalcoholic disease (NAFLD). Although the exact molecular pathway leading to impaired insulin signaling has not been definitively established, ceramides are suspected mediators of lipid induced hepatic insulin resistance. Therefore, the aim of the study was to evaluate the serum ceramides concentration in obese children with NAFLD.The prospective study included 80 obese children (aged 7-17\xa0years, median 12\xa0years) admitted to our Department to diagnose initially suspected disease. Patients with viral hepatitis (HCV, HBV, CMV), autoimmune (AIH), toxic and metabolic (Wilson\'s disease, alfa-1-antitrypsin deficiency) diseases and celiac disease were excluded. NAFLD was diagnosed based on pediatric diagnostic criteria in obese children with steatosis in ultrasound (US) as well as elevated alanine transaminase (ALT) serum activity after exclusion of other major diseases listed before. Ultrasonography was used as a screening method and for qualitative assessment of the steatosis degree (graded according to Saverymuttu scale). Advanced steatosis was defined as a score\u2009>\u20091. The total intrahepatic lipid content (TILC) was assessed by magnetic resonance proton spectroscopy (HMRS) which is the most accurate technique for assessment of ectopic fat accumulation. Fasting serum concentration of ceramides was measured in 62 children.NAFLD was diagnosed in 31 children. Significant, positive correlation was found between total serum concentration of ceramides and insulin (r\xa0=\u20090.3, p\xa0=\u20090.02) and HOMA-IR (r\xa0=\u20090.28, p\xa0=\u20090.03). Total ceramide concentration as well as specific -ceramides (FA-ceramides) concentrations, namely: myristic, , palmitoleic, stearic, oleic, behenic and lignoceric were significantly higher (p\xa0=\u20090.004, p\xa0=\u20090.003, p\xa0=\u20090.007, p\xa0<\u20090.001, p\xa0=\u20090.035, p\xa0=\u20090.008, p\xa0=\u20090.003, p\xa0=\u20090.006, respectively) in children with NAFLD compared to controls (n\xa0=\u200914). Moreover, children with NAFLD had significantly higher activity of ALT (p\xa0<\u20090.001) and GGT (p\xa0<\u20090.001), HOMA-IR (p\xa0=\u20090.04), BMI (p\xa0=\u20090.046), waist circumference (p\xa0=\u20090.01) steatosis grade in ultrasound (p\xa0<\u20090.001) and TILC in HMRS (p\xa0<\u20090.001) compared to children without NAFLD. We did not find significant differences in total and FA-ceramide species concentrations between children with mild (grade 1) and advanced steatosis in ultrasonography (grade 2-3).Elevated ceramide concentrations in obese patients together with their significant correlation with insulin resistance parameters suggest their association with molecular pathways involved in insulin signaling impairment known to be strongly linked to pathogenesis of non-alcoholic disease.

Keyword: fatty liver

Protein translation associated to PERK arm is a new target for regulation of metainflammation: A connection with hepatocyte cholesterol.

Endoplasmic reticulum stress is a cellular phenomenon that has been associated with metabolic disorders, contributing to the development of obesity, disease, and dyslipidemias. Under metabolic overload conditions, in cells with a high protein-secretory activity, such as hepatocytes and Langerhans β cells, the unfolded protein response (UPR) is critical in to maintain protein homeostasis (proteostasis). UPR integrated by a tripartite signaling system, through activating transcription factor 6, protein kinase R-like endoplasmic reticulum kinase (PERK), and inositol-requiring enzyme 1, regulates gene transcription and translation to resolve stress and conserve proteostasis. In the current study, we demonstrated in hepatocytes under metabolic overload by saturated and stearic acids, through activation of PERK signaling and CCAAT-enhancer-binding protein homologous protein (CHOP) transcription factor, an association with the expression of cyclooxygenase 2. More important, isolated exosomes from supernatants of macrophages exposed to lipopolysaccharides can also induce a metainflammation phenomenon, and when\xa0treated on hepatocytes, induced a rearrangement in cholesterol metabolism through sterol regulatory element-binding protein 2 (SREBP2), low-density lipoprotein receptor (LDLR), apolipoprotein A-I, and ABCA1. Moreover, we demonstrate the cellular effect of terpene-derived molecules, such as cryptotanshinone, isolated of plant Salvia brandegeei, regulating metainflammatory conditions through PERK pathway in both hepatocytes and β cells. Our data suggest the presence of a modulatory mechanism on specific protein translation process. This effect could be mediated by eukaryotic initiation factor-4A, evaluating salubrinal as a control molecule. Likewise, the protective mechanisms of unsaturated acids, such as oleic and palmitoleic were confirmed. Therefore, modulation of metainflammation suggests a new target through PERK signaling in cells with a high secretory activity, and possibly the regulation of cholesterol in hepatocytes is promoted via exosomes.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Melatonin improves insulin resistance and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with insulin resistance and type 2 diabetes. The aim of this study was to determine whether melatonin improves hepatic insulin resistance and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve insulin resistance in FETUA-overexpressing cells, indicating that improvement in insulin resistance by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced body weight gain to one-third of that of the HFD group and hepatic steatosis. Insulin sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic insulin resistance and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: fatty liver

The BACE1 product sAPPβ induces ER stress and inflammation and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, insulin resistance, PGC-1α down-regulation, and oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and insulin resistance in skeletal muscle and gluconeogenesis in .Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Chronic intake of moderate fat-enriched diet induces and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Disease (NAFLD) is the cause of chronic disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate -- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and function. samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced . We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: fatty liver

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic (NAFL) to non-alcoholic steatohepatitis (NASH), cirrhosis, and cancer. The role of different types of acids in driving the early progression of NAFL to NASH is not understood. Lipid overload causing lipotoxicity and inflammation has been considered as an essential pathogenic factor. To correlate the lipid profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced lipid overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced inflammation and cell death, whereas C16:1 induced significant lipid droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. Lipid profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific acids that affect lipid metabolism and cause lipotoxicity or lipid droplet formation. It indicates that not only saturation or monounsaturation of acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of lipid metabolism and the heterogeneous development of NAFLD.

Keyword: fatty liver

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while collagen content was determined by Sirius Red staining. Both interesterified fats increased collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: fatty liver

Dynamic alterations in the gut microbiota and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut microbiota and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut microbiota dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut microbiota and metabolome.

Keyword: fatty liver

Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high-fat diet-fed mice.

Type 2 diabetes mellitus is frequently accompanied by disease. Lipid accumulation within the is considered as one of the risk factors for insulin resistance. Hepatocyte growth factor (HGF) is used to treat dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting insulin and homeostatic model assessment of insulin resistance were calculated for insulin sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/ synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic insulin resistance and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

C-X-C Motif Chemokine 10 Impairs Autophagy and Autolysosome Formation in Non-alcoholic Steatohepatitis.

C-X-C motif chemokine 10 (CXCL10) is a crucial pro-inflammatory factor in chronic hepatitis. Autophagy dysregulation is known to contribute to hepatic inflammatory injury. Hence, we investigated the regulatory effect of CXCL10 on the autophagosome-lysosome system during non-alcoholic disease (NAFLD) development. The effect of CXCL10 ablation by neutralizing monoclonal antibody (mAb) or genetic knockout on autophagic flux was evaluated in cultured hepatocytes and animal models of NAFLD. Results demonstrated that CXCL10 ablation protected against hepatocyte injury and steatohepatitis development in mice. Autophagic flux impairment was rectified by CXCL10 inhibition using anti-CXCL10 mAb in AML-12 and HepG2 cell lines and primary hepatocytes as evidenced by the attenuated accumulation of p62/SQSTM1 and LC3-II proteins and increased autophagic protein degradation. Impaired autophagic flux was significantly restored by CXCL10 knockout or anti-CXCL10 mAb in mice. Bafilomycin A1, an inhibitor of autolysosome formation, abolished the rectifying effect of anti-CXCL10 mAb or CXCL10 knockdown in AML-12 and primary hepatocytes, indicating CXCL10 impaired late-stage autophagy in NAFLD. Anti-CXCL10 mAb treatment also increased the fusion of LC3-positive autophagosomes with lysosomes in HepG2 cells challenged with , suggesting that CXCL10 ablation restored autolysosome formation. Consistently, the number of autolysosomes was significantly increased by CXCL10 knockout in mice as shown by electron microscopy. In conclusion, upregulated CXCL10 in steatohepatitis impairs autophagic flux by reducing autolysosome formation, thereby inhibiting autophagic protein degradation and the accumulation of ubiquitinated proteins, leading to the development of steatohepatitis.

Keyword: fatty liver

Lipotoxicity and the gut- axis in NASH pathogenesis.

The pathogenesis of non-alcoholic disease, particularly the mechanisms whereby a minority of patients develop a more severe phenotype characterised by hepatocellular damage, inflammation, and fibrosis is still incompletely understood. Herein, we discuss two pivotal aspects of the pathogenesis of NASH. We first analyse the initial mechanisms responsible for hepatocellular damage and inflammation, which derive from the toxic effects of excess lipids. Accumulating data indicate that the total amount of triglycerides stored in hepatocytes is not the major determinant of lipotoxicity, and that specific lipid classes act as damaging agents on cells. In particular, the role of free acids such as , cholesterol, lysophosphatidylcholine and ceramides has recently emerged. These lipotoxic agents affect the cell behaviour via multiple mechanisms, including activation of signalling cascades and death receptors, endoplasmic reticulum stress, modification of mitochondrial function, and oxidative stress. In the second part of this review, the cellular and molecular players involved in the cross-talk between the gut and the are considered. These include modifications to the microbiota, which provide signals through the intestine and bacterial products, as well as hormones produced in the bowel that affect metabolism at different levels including the . Finally, the activation of nuclear receptors by bile acids is analysed.Copyright © 2017. Published by Elsevier B.V.

Keyword: fatty liver

Attenuated lipotoxicity and apoptosis is linked to exogenous and endogenous augmenter of regeneration by different pathways.

Nonalcoholic disease (NAFLD) covers a spectrum from simple steatosis to nonalcoholic steatohepatitis (NASH) and cirrhosis. Free acids (FFA) induce steatosis and lipo-toxicity and correlate with severity of NAFLD. In this study we aimed to investigate the role of exogenous and endogenous ALR (augmenter of regeneration) for FFA induced ER (endoplasmatic reticulum) -stress and lipoapoptosis. Primary human hepatocytes or hepatoma cells either treated with recombinant human ALR (rhALR, 15kDa) or expressing short form ALR (sfALR, 15kDa) were incubated with (PA) and analyzed for lipo-toxicity, -apoptosis, activation of ER-stress response pathways, triacylglycerides (TAG), mRNA and protein expression of lipid metabolizing genes. Both, exogenous rhALR and cytosolic sfALR reduced PA induced caspase 3 activity and Bax protein expression and therefore lipotoxicity. Endogenous sfALR but not rhALR treatment lowered TAG levels, diminished activation of ER-stress mediators C-Jun N-terminal kinase (JNK), X-box binding protein-1 (XBP1) and proapoptotic transcription factor C/EBP-homologous protein (CHOP), and reduced death receptor 5 protein expression. Cellular ALR exerts its lipid lowering and anti-apoptotic actions by enhancing FABP1, which binds toxic FFA, increasing mitochondrial β-oxidation by elevating the mitochondrial FFA transporter CPT1α, and decreasing ELOVL6, which delivers toxic FFA metabolites. We found reduced hepatic mRNA levels of ALR in a high fat diet mouse model, and of ALR and FOXA2, a transcription factor inducing ALR expression, in human steatotic as well as NASH samples, which may explain increased lipid deposition and reduced β-oxidation in NASH patients. Present study shows that exogenous and endogenous ALR reduce PA induced lipoapoptosis. Furthermore, cytosolic sfALR changes mRNA and protein expression of genes regulating lipid metabolism, reduces ER-stress finally impeding progression of NASH.

Keyword: fatty liver

Anticandidal activity of the extract and compounds isolated from Rottb.

The phytochemical screening of showed that carbohydrates and/or glycosides, flavonoids, tannins, sterols and/or triterpenes, and proteins and/or amino acids are present. The profile comprised major; , oleic, heptadecanoic, linoleic and minor; arachidonic, lignoceric, stearic, and myristic . Two compounds; namely, α-amyrin and β-sitosterol were isolated by the fractionation of unsaponifiable matter. The acute toxicity study showed that the reported after oral administration of the alcohol extract ( showed that the plant was highly safe as the LD was more than 4000\u202fmg/kg. These results were well supported by the sub-chronic toxicity, as the administrated to rats for 15 consecutive days at dose 1000\u202fmg/kg showed no alteration in the and kidney functions. Moreover, the extract of the plant exhibited anti-candidal activity against different species. The most potent activity, (23.1\u202f±\u202f2.1, 0.98\u202fµg/ml) and (22.3\u202f±\u202f0.53, 0.98\u202fµg/ml), was obtained by the chloroform and total extract, respectively against .

Keyword: fatty liver

ACSL5 genotype influence on metabolism: a cellular, tissue, and whole-body study.

Acyl-CoA Synthetase Long Chain 5 (ACSL5) gene\'s rs2419621 T/C polymorphism was associated with ACSL5 mRNA expression and response to lifestyle interventions. However, the mechanistic understanding of the increased response in T allele carriers is lacking. Study objectives were to investigate the effect of rs2419621 genotype and ACSL5 human protein isoforms on oxidation and respiration.Human ACSL5 overexpression in C2C12 mouse myoblasts was conducted to measure C oxidation and protein isoform localization in vitro. C oxidation studies and Western blot analysis of ACSL5 proteins were carried out in rectus abdominis primary myotubes from 5 rs2419621 T allele carriers and 4 non-carriers. In addition, mitochondrial high-resolution respirometry was conducted on vastus lateralis muscle biopsies from 4 rs2419621 T allele carriers and 4 non-carriers. Multiple linear regression analysis was conducted to test the association between rs2419621 genotype and respiratory quotient related pre- and post-lifestyle intervention measurements in postmenopausal women with overweight or obesity.In comparison to rs2419621 non-carriers, T allele carriers displayed higher levels of i) 683aa ACSL5 isoform, localized mainly in the mitochondria, playing a greater role in oxidation in comparison to the 739aa protein isoform ii) in vitro CO production in rectus abdominis primary myotubes iii) in vivo oxidation and lower carbohydrate oxidation post-intervention iv) ex vivo complex I and II tissue respiration in vastus lateralis muscle.These results support the conclusion that rs2419621 T allele carriers, are more responsive to lifestyle interventions partly due to an increase in the short ACSL5 protein isoform, increasing cellular, tissue and whole-body utilization. With the increasing effort to develop personalized medicine to combat obesity, our findings provide additional insight into genotypes that can significantly affect whole body metabolism and response to lifestyle interventions.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Loss of toll-like receptor 3 aggravates hepatic inflammation but ameliorates steatosis in mice.

The importance of toll-like receptor (TLR) 4 in the pathogenesis of steatohepatitis has been well documented; however, little is known about the role of TLR3. In this study, we determined whether the depletion of TLR3 modulated hepatic injury in mice and further aimed to provide mechanistic insights into the TLR3-mediated modulation of diet-induced hepatic inflammation and fat accumulation. Hepatic steatosis and inflammatory response were induced by feeding wild-type (WT) or TLR3 knockout mice a high-fat diet for 8 weeks. Primary resident cells, including hepatocytes, Kupffer cells, and hepatic stellate cells (HSCs), were treated with . TLR3 knockout mice fed a high-fat diet showed severe hepatic inflammation accompanied by nuclear factor-κB and IRF3 activation, which is mainly induced by the activation of Kupffer cells. Decreased TLR4 expression was restored in hepatic mononuclear cells and Kupffer cells in TLR3 knockout mice compared to that in the WT. Moreover, hepatic steatosis was decreased in TLR3 knockout mice. Hepatocytes from TLR3 knockout mice exhibited reduced expression of cannabinoid receptors. HSCs from TLR3 knockout mice showed decreased expression of the enzymes involved in endocannabinoid synthesis. In conclusion, this study suggests that the selective modulation of TLR3 could be a novel therapeutic target for the treatment of hepatic inflammation and steatosis.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Tannic , a novel histone acetyltransferase inhibitor, prevents non-alcoholic disease both in\xa0vivo and in\xa0vitro model.

We examined the potential of tannic (TA) as a novel histone acetyltransferase inhibitor (HATi) and demonstrated that TA prevents non-alcoholic disease (NAFLD) by inhibiting HAT activity.The anti-HAT activity of TA was examined using HAT activity assays. An in\xa0vitro NAFLD model was generated by treating HepG2 cells with oleic and acids. Male C57BL/6J mice were fed a control diet (CD) or Western diet (WD) with or without supplementation with either 1% or 3% TA (w/w) for 12 weeks. Finally, the possibility of interacting p300 and TA was simulated.TA suppressed HAT activity both in\xa0vitro and in\xa0vivo. Interestingly, TA abrogated occupancy of p300 on the sterol regulatory element in the synthase and ATP-citrate lyase promoters, eventually inducing hypoacetylation of H3K9 and H3K36. Furthermore, TA decreased acetylation at lysine residues 9 and 36 of histone H3 protein and that of total proteins. Consequently, TA decreased the mRNA expression of lipogenesis-related genes and attenuated lipid accumulation in\xa0vivo. We observed that NAFLD features, including body weight, mass, fat mass, and lipid profile in serum, were improved by TA supplementation in\xa0vivo. Finally, we demonstrated the possibility that TA directly binds to p300 through docking simulation between ligand and protein.Our findings demonstrate that TA, a novel HATi, has potential application for the prevention of NAFLD.Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: fatty liver

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive oxygen species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect inflammation and lipid content in the .The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and inflammation induced by HF diet , leading to improved metabolic disorder and associated injury in a mouse model of non-alcoholic disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: fatty liver

Role of 3-Hydroxy -Induced Hepatic Lipotoxicity in Acute of Pregnancy.

Acute of pregnancy (AFLP), a catastrophic illness for both the mother and the unborn offspring, develops in the last trimester of pregnancy with significant maternal and perinatal mortality. AFLP is also recognized as an obstetric and medical emergency. Maternal AFLP is highly associated with a fetal homozygous mutation (1528G>C) in the gene that encodes for mitochondrial long-chain hydroxy acyl-CoA dehydrogenase (LCHAD). The mutation in LCHAD results in the accumulation of 3-hydroxy acids, such as 3-hydroxy myristic , 3-hydroxy and 3-hydroxy dicarboxylic in the placenta, which are then shunted to the maternal circulation leading to the development of acute injury observed in patients with AFLP. In this review, we will discuss the mechanistic role of increased 3-hydroxy in causing lipotoxicity to the and in inducing oxidative stress, mitochondrial dysfunction and hepatocyte lipoapoptosis. Further, we also review the role of 3-hydroxy acids in causing placental damage, pancreatic islet β-cell glucolipotoxicity, brain damage, and retinal epithelial cells lipoapoptosis in patients with LCHAD deficiency.

Keyword: fatty liver

Lycopus lucidus Turcz. ex Benth. Attenuates free -induced steatosis in HepG2 cells and non-alcoholic disease in high-fat diet-induced obese mice.

Non-alcoholic disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the . Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fatty liver

High-fat diet promotes experimental by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) tight junction regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes in a 2,4,6-trinitrobenzenesulfonic -induced experimental model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: fatty liver

Involvement of sarco/endoplasmic reticulum calcium ATPase-mediated calcium flux in the protective effect of oleic against lipotoxicity in hepatocytes.

Elevated free acids, particularly saturated ones such as , may play an important role in the lipotoxic mechanism of nonalcoholic disease (NAFLD). Saturated acids induce autophagy dysfunction and endoplasmic reticulum (ER) stress leading to apoptosis in hepatocytes. However, unsaturated acids, such as oleic , are nontoxic and can even prevent saturated -induced toxicity in vitro. Although emerging evidence has suggested that ER calcium flux disruption in hepatocytes is involved in NAFLD pathogenesis, the roles of acids in autophagy and ER calcium flux still remain unclear. We demonstrated that oleic ameliorated -induced autophagy arrest and ER stress in parallel with ER calcium depletion in hepatocytes. Moreover, we found that the effect of oleic against autophagy arrest was reversed by the pharmacological inhibition of sarcoplasmic reticulum Ca-ATPase (SERCA), which influxes calcium to ER. These data suggest that SERCA-mediated ER calcium flux is greatly involved in -induced lipotoxicity in hepatocytes, and the prevention of ER calcium depletion may restore saturated -induced autophagy arrest in hepatocytes.Copyright © 2019. Published by Elsevier Inc.

Keyword: fatty liver

Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Impact of miR-140 Deficiency on Non-Alcoholic Disease.

We have previously shown that loss of miR-140 has a pro-fibrotic effect in the mammary gland. This study aims to investigate whether miR-140 loss and obesity act synergistically to promote non-alcoholic disease (NAFLD), and to identify the underlying mechanisms. tissues were isolated from lean-fat-diet and high-fat-diet fed wild-type and miR-140 knockout mice. Using molecular staining and immunohistochemistry techniques, increased development of NAFLD and fibrotic indicators in miR-140 knockout mice were identified. Utilizing an in vitro model system, miR-140 was demonstrated to target TLR-4, and miR-140 overexpression was shown to be sufficient to inhibit signaling through the TLR-4/NFκB pathway.These findings demonstrate that loss of miR-140 results in increased expression of TLR-4, sensitizing cells to signaling and in increased inflammatory activity through the TLR4/NFκB pathway. This signaling axis promotes NAFLD development in a high-fat diet context and indicates the potential utility of miR-140 rescue as a therapeutic strategy in NAFLD.© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

Hydroxyhydroquinone impairs fat utilization in mice by reducing nitric oxide availability.

Habitual consumption of chlorogenic compounds (CGAs) from coffee increases fat catabolism and reduces body fat; however, the contribution of roasted coffee remains unclear. Hydroxyhydroquinone (HHQ) impairs the vasodilatory and antihypertensive effects of CGAs by reducing nitric oxide (NO) bioavailability. Since HHQ also reduces fat catabolism, we hypothesized that HHQ does so by decreasing NO availability. Therefore, we investigated the effect of HHQ on energy metabolism in KKAy mice. In HHQ-treated mice, fat oxidation was significantly low and dose-dependent, serum and urinary hydrogen peroxide were high, and plasma NO metabolites and S-nitrosylated proteins were low. In HHQ-treated mouse hepatocytes, the palmitate-induced increase in cellular oxygen consumption was negatively affected, and HHQ or L-NAME reduced cellular utilization. In conclusion, HHQ can impair fat utilization by reducing NO availability in mice. Protein S-nitrosylation reduction in cells after HHQ consumption may be associated with impaired oxidation.

Keyword: fatty liver

PPARγ alleviated hepatocyte steatosis through reducing SOCS3 by inhibiting JAK2/STAT3 pathway.

Peroxisome proliferator-activated receptor gamma (PPARγ) participates in the process of insulin resistance (IR), a crucial pathophysiology in non-alcoholic disease (NAFLD). Meanwhile, suppressor of cytokine signaling3 (SOCS3) also regulates IR in NAFLD. Both PPARγ and SOCS3 play a role in NAFLD through regulating IR, while it is unclear whether these two proteins interact to regulate hepatic steatosis. PPARγ, SOCS3 and its associated JAK2/STAT3 pathway were analyzed using Kuppfer cells (KCs) treatment with LPS and BRL-3A cells treatment with , KC-conditioned medium (KCCM), PPARγ agonist rosiglitazone (ROZ) or JAK2 inhibitor AG490 to demonstrate the role of PPARγ and SOCS3 in hepatocytes steatosis. As LPS concentration increasing, phagocytosis activity of KCs decreased; but releasing of TNF-α and IL-6 increased. After treatment with KCCM, mRNA level of SOCS3, JAK2 and STAT3 as well as protein expression of SOCS3, p-JAK2 and p-STAT3 in steatosis BRL-3A cells increased significantly, which were inhibited by AG490 or ROZ treatment. Taken together, these results indicated that KCCM attributed to KCs dysfunction facilitated hepatocyte steatosis through promoting expressing SOCS3; but PPARγ agonist ROZ alleviated steatosis through reducing SOCS3 expression by inhibiting JAK2/STAT3 in hepatocytes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Bromide alleviates -induced lipid accumulation in mouse primary hepatocytes through the activation of PPARα signals.

Increased plasma free acids (FFAs) and triglyceride (TG) accumulations have been implicated in the pathogenesis of hepatic steatosis. On the other hand, trace elements function as essential cofactors that are involved in various biochemical processes in mammals, including metabolic homeostasis. Notably, clinical and animal studies suggest that the plasma levels of bromide negatively correlate with those of TG, total cholesterol (TC) and high-density lipoprotein-cholesterol (HDL-C). However, the effect of bromide on lipid accumulation and the direct molecular target responsible for its action remains unknown. Oil red O (ORO) and Nile red staining were used to detect the effect of bromide on lipid accumulation in mouse primary hepatocytes (PHs) treated with different doses of sodium bromide (NaBr) in the presence of FFAs (0.4\xa0mM oleate/ 1:1). Spectrophotometric and fluorometric analyses were performed to assess cellular TG concentrations and rates of oxidation (FAO), respectively, in mouse PHs. We found that bromide decreased FFA-induced lipid accumulation and increased FFA-inhibited oxygen consumptions in mouse PHs in a dose-dependent manner via activation of PPARα. Mechanical studies demonstrated that bromide decreased the phosphorylation levels of JNK. More importantly, the PPARα-specific inhibitor GW6471 partially abolished the beneficial effects of bromide on mouse PHs. Bromide alleviates FFA-induced excessive lipid storage and increases rates of FAO through the activation of PPARα/JNK signals in mouse PHs. Therefore, bromide may serve as a novel drug in the treatment of hepatic steatosis.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

miR-1224-5p Enhances Hepatic Lipogenesis by Targeting Adenosine Monophosphate-Activated Protein Kinase α1 in Male Mice.

MicroRNAs are potential therapeutic targets for metabolic diseases. Here, miR-1224-5p was highly expressed in the livers of mice fed a high-fat diet (HFD) and in obese (ob/ob) mice. To examine the potential role of miR-1224-5p, we constructed -specific adenoviral vectors expressing either an miR-1224-5p inhibitor sequence or miR-1224-5p mimic sequences. After tail-vein vector injection, HFD-fed mice were examined for expression of lipogenic genes. We found that miR-1224-5p inhibitors significantly attenuated hepatic lipogenesis and steatosis in HFD-fed mice, whereas miR-1224-5p mimicked promoted lipid accumulation in the of chow-fed C57BL/6 mice. Additional in vitro studies demonstrated that downregulation of miR-1224-5p in HepG2 and primary hepatocytes led to a reduction of cellular triglycerides after treatment with an oleic and mixture. Importantly, this study also identified adenosine monophosphate-activated protein kinase (AMPK)-α1 as a direct target of miR-1224-5p. miR-1224-5p binding to the 3\' untranslated region of AMPKα1 suppressed expression of the AMPKα1 protein and its downstream molecules. Metformin, an activator of AMPK, also inhibited hepatic expression of miR-1224-5p. Together, these findings indicate that miR-1224-5p promotes hepatic lipogenesis by suppressing AMPKα1 expression and suggest that miR-1224-5p inhibitors warrant further investigation as potential therapeutic tools in the treatment of nonalcoholic disease.

Keyword: fatty liver

Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4-MD2 complex.

Reactive oxygen species (ROS) contribute to the development of non-alcoholic disease. ROS generation by infiltrating macrophages involves multiple mechanisms, including Toll-like receptor 4 (TLR4)-mediated NADPH oxidase (NOX) activation. Here, we show that palmitate-stimulated CD11bF4/80 hepatic infiltrating macrophages, but not CD11bF4/80 Kupffer cells, generate ROS via dynamin-mediated endocytosis of TLR4 and NOX2, independently from MyD88 and TRIF. We demonstrate that differently from LPS-mediated dimerization of the TLR4-MD2 complex, palmitate binds a monomeric TLR4-MD2 complex that triggers endocytosis, ROS generation and increases pro-interleukin-1β expression in macrophages. Palmitate-induced ROS generation in human CD68CD14 macrophages is strongly suppressed by inhibition of dynamin. Furthermore, Nox2-deficient mice are protected against high-fat diet-induced hepatic steatosis and insulin resistance. Therefore, endocytosis of TLR4 and NOX2 into macrophages might be a novel therapeutic target for non-alcoholic disease.

Keyword: fatty liver

Hepatocyte nuclear receptor SHP suppresses inflammation and fibrosis in a mouse model of nonalcoholic steatohepatitis.

Nonalcoholic disease (NAFLD) is a burgeoning health problem worldwide, ranging from nonalcoholic (NAFL, steatosis without hepatocellular injury) to the more aggressive nonalcoholic steatohepatitis (NASH, steatosis with ballooning, inflammation, or fibrosis). Although many studies have greatly contributed to the elucidation of NAFLD pathogenesis, the disease progression from NAFL to NASH remains incompletely understood. Nuclear receptor small heterodimer partner (Nr0b2, ) is a transcriptional regulator critical for the regulation of bile , glucose, and lipid metabolism. Here, we show that SHP levels are decreased in the livers of patients with NASH and in diet-induced mouse NASH. Exposing primary mouse hepatocytes to and lipopolysaccharide , we demonstrated that the suppression of expression in hepatocytes is due to c-Jun N-terminal kinase (JNK) activation, which stimulates c-Jun-mediated transcriptional repression of Interestingly, induction of hepatocyte-specific SHP in steatotic mouse ameliorated NASH progression by attenuating inflammation and fibrosis, but not steatosis. Moreover, a key mechanism linking the anti-inflammatory role of hepatocyte-specific SHP expression to inflammation involved SHP-induced suppression of NF-κB p65-mediated induction of chemokine (C-C motif) ligand 2 (CCL2), which activates macrophage proinflammatory polarization and migration. In summary, our results indicate that a JNK/SHP/NF-κB/CCL2 regulatory network controls communications between hepatocytes and macrophages and contributes to the disease progression from NAFL to NASH. Our findings may benefit the development of new management or prevention strategies for NASH.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: fatty liver

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in . Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: fatty liver

Skeletal muscle miR-34a/SIRT1:AMPK axis is activated in experimental and human non-alcoholic steatohepatitis.

Non-alcoholic disease (NAFLD) pathogenesis associates with intramyocellular lipid deposition and mitochondrial dysfunction. microRNAs (miRs), including pro-apoptotic miR-34a, are modulated during disease progression in tissue and plasma. We aimed to investigate the functional role of the miR-34a/SIRT1:AMP-activated protein kinase (AMPK) pathway in modulating local mitochondrial dysfunction in the skeletal muscle of human and experimental non-alcoholic steatohepatitis. Muscle biopsies were obtained from morbid obese NAFLD patients undergoing bariatric surgery. C57BL/6N mice were fed different NAFLD-inducing diets and C2C12 muscle cells incubated with (PA) in the presence or absence of an AMPK activator, or upon miR-34a functional modulation. Several muscle miRNAs, including miR-34a, were found increased with human NAFLD progression. Activation of the miR-34a/SIRT1:AMPK pathway, concomitant with impairment in insulin signalling mediators and deregulation of mitochondrial-shaping proteins, was evident in C2C12 cells incubated with PA, as well as in the skeletal muscle of all three diet-induced NAFLD mice models. Functional studies established the association between miR-34a- and PA-induced muscle cell deregulation. Of note, activation of AMPK almost completely prevented miR-34a- and PA-induced cellular stress. In addition, the miR-34a/SIRT1:AMPK pathway and mitochondrial dynamics dysfunction were also found amplified in muscle of human NAFLD. Finally, muscle miR-34a expression and mitofusin 2 (Mfn2) protein levels correlated with hallmarks of NAFLD and disease progression. Our results indicate that activation of the miR-34a/SIRT1:AMPK pathway leads to mitochondrial dynamics dysfunction in skeletal muscle of human and experimental NAFLD, representing an appealing prospective target in metabolic syndrome. KEY MESSAGES: Skeletal muscle microRNAs are modulated during NAFLD progression. -induced muscle cell dysfunction occurs, at least in part, through activation of the miR-34a/SIRT1:AMPK pathway. miR-34a/SIRT1:AMPK activation associates with mitochondria dynamics dysfunction in human NAFLD.

Keyword: fatty liver

Moderate chronic ethanol consumption exerts beneficial effects on nonalcoholic in mice fed a high-fat diet: possible role of higher formation of triglycerides enriched in monounsaturated acids.

Several clinical studies suggested that light-to-moderate alcohol intake could alleviate nonalcoholic disease (NAFLD), but the underlying mechanism is still poorly understood.Mice fed a high-fat diet (HFD) were submitted or not to moderate ethanol intake for 3\xa0months (ca. 10\xa0g/kg/day) via drinking water. Biochemical, analytical and transcriptomic analyses were performed in serum and .Serum ethanol concentrations in ethanol-treated HFD mice comprised between 0.5 and 0.7\xa0g/l throughout the experiment. NAFLD improvement was observed in ethanol-treated HFD mice as assessed by reduced serum transaminase activity. This was associated with less microvesicular and more macrovacuolar steatosis, the absence of apoptotic hepatocytes and a trend towards less fibrosis. lipid analysis showed increased amounts of acids incorporated in triglycerides and phospholipids, reduced proportion of in total lipids and higher desaturation index, thus suggesting enhanced stearoyl-coenzyme A desaturase activity. mRNA expression of several glycolytic and lipogenic enzymes was upregulated. Genome-wide expression profiling and gene set enrichment analysis revealed an overall downregulation of the expression of genes involved in collagen fibril organization and leukocyte chemotaxis and an overall upregulation of the expression of genes involved in oxidative phosphorylation and mitochondrial respiratory chain complex assembly. In addition, mRNA expression of several proteasome subunits was upregulated in ethanol-treated HFD mice.Moderate chronic ethanol consumption may alleviate NAFLD by several mechanisms including the generation of non-toxic lipid species, reduced expression of profibrotic and proinflammatory genes, restoration of mitochondrial function and possible stimulation of proteasome activity.

Keyword: fatty liver

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the . Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid metabolism and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic disease.

Keyword: fatty liver

Apigenin reduces the excessive accumulation of lipids induced by via the AMPK signaling pathway in HepG2 cells.

In recent years, increasing attention has been paid to diseases caused by excessive accumulation of lipids in the with therapeutic agents derived from natural products offering an alternative treatment to conventional therapies. Among these therapeutic agents, apigenin, a natural flavonoid, has been proven to exert various beneficial biological effects. In the present study, the antiadipogenic effects of apigenin in HepG2 cells was investigated. It was demonstrated that the treatment of cells with different concentrations of apigenin for 24 h significantly decreased the -induced increases in total cholesterol (TC) and triglyceride (TG) levels as well as intracellular lipid accumulation. In addition, apigenin increased the phosphorylated-AMP-activated protein kinase (AMPK) levels but decreased the expression levels of 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding protein (SREBP)-1, synthase, and SREBP-2 in a concentration-dependent manner. The present findings suggested that apigenin might improve lipid metabolism by activating the AMPK/SREBP pathway to reduce lipid accumulation in the .Copyright: © Lu et al.

Keyword: fatty liver

Role of caveolin-1 in hepatocellular carcinoma arising from non-alcoholic disease.

The molecular features of hepatocellular carcinoma arising from non-alcoholic disease (NAFLD-HCC) are not well known. In this study, we investigated the mechanism by which NAFLD-HCC survives in a fat-rich environment. We found that caveolin (CAV)-1 was overexpressed in clinical specimens from NAFLD-HCC patients. HepG2, HLE, and HuH-7 HCC cell lines showed decreased proliferation in the presence of the saturated acids and stearic , although only HLE cells expressed high levels of CAV-1. HLE cells treated with oleic (OA) showed robust proliferation, whereas CAV-null HepG2 cells showed reduced proliferation and increased apoptosis. CAV-1 knockdown in HLE cells attenuated the OA-induced increase in proliferation and enhanced apoptosis. Liquid chromatography-tandem mass spectrometry analysis revealed that the levels of OA-containing ceramide, a pro-apoptotic factor, were higher in HepG2 and CAV-1-deficient HLE cells than in HLE cells, suggesting that CAV-1 inhibits apoptosis by decreasing the level of OA-containing ceramide. These results indicate that CAV-1 is important for NAFLD-HCC survival in -rich environments and is a potential therapeutic target.© 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

Keyword: fatty liver

Ground Beef High in Total Fat and Saturated Acids Decreases X Receptor Signaling Targets in Peripheral Blood Mononuclear Cells of Men and Women.

We hypothesized that consumption of saturated acids in the form of high-fat ground beef for 5 weeks would depress X receptor signaling targets in peripheral blood mononuclear cells (PBMC) and that changes in gene expression would be associated with the corresponding changes in lipoprotein cholesterol (C) concentrations. Older men (n = 5, age 68.0 ± 4.6 years) and postmenopausal women (n = 7, age 60.9 ± 3.1 years) were assigned randomly to consume ground-beef containing 18% total fat (18F) or 25% total fat (25F), five patties per week for 5 weeks with an intervening 4-week washout period. The 25F and 18F ground-beef increased (p < 0.05) the intake of saturated fat, monounsaturated fat, , and stearic , but the 25F ground-beef increased only the intake of oleic (p < 0.05). The ground-beefs 18F and 25F increased the plasma concentration of (p < 0.05) and decreased the plasma concentrations of arachidonic, eicosapentaenoic, and docosahexaenic acids (p < 0.05). The interventions of 18F and 25F ground-beef decreased very low-density lipoprotein C concentrations and increased particle diameters and low-density lipoprotein (LDL)-I-C and LDL-II-C concentrations (p < 0.05). The ground-beef 25F decreased PBMC mRNA levels for the adenosine triphosphate (ATP) binding cassette A, ATP binding cassette G1, sterol regulatory element binding protein-1, and LDL receptor (LDLR) (p < 0.05). The ground-beef 18F increased mRNA levels for stearoyl-CoA desaturase-1 (p < 0.05). We conclude that the increased LDL particle size and LDL-I-C and LDL-II-C concentrations following the 25F ground-beef intervention may have been caused by decreased hepatic LDLR gene expression.© 2018 AOCS.

Keyword: fatty liver

Picroside II attenuates fatty accumulation in HepG2 cells via modulation of fatty uptake and synthesis.

Hepatic steatosis is caused by an imbalance between free fatty acids (FFAs) uptake, utilization, storage, and disposal. Understanding the molecular mechanisms involved in FFAs accumulation and its modulation could drive the development of potential therapies for Nonalcoholic fatty liver disease. The aim of the current study was to explore the effects of picroside II, a phytoactive found in , on fatty accumulation vis-à-vis silibinin, a known hepatoprotective phytoactive from .HepG2 cells were loaded with FFAs (oleic :/2:1) for 20 hours to mimic hepatic steatosis. The FFAs concentration achieving maximum fat accumulation and minimal cytotoxicity (500 μM) was standardized. HepG2 cells were exposed to the standardized FFAs concentration with and without picroside II pretreatment.Picroside II pretreatment inhibited FFAs-induced lipid accumulation by attenuating the expression of fatty transport protein 5, sterol regulatory element binding protein 1 and stearoyl CoA desaturase. Preatreatment with picroside II was also found to decrease the expression of forkhead box protein O1 and phosphoenolpyruvate carboxykinase.These findings suggest that picroside II effectively attenuated fatty accumulation by decreasing FFAs uptake and lipogenesis. Picroside II also decreased the expression of gluconeogenic genes.

Keyword: gluconeogenesis

CsrA regulates a metabolic switch from amino to glycerolipid metabolism.

CsrA plays a crucial role in the life-stage-specific expression of virulence phenotypes and metabolic activity. However, its exact role is only partly known. To elucidate how CsrA impacts metabolism we analysed the CsrA depended regulation of metabolic functions by comparative C-isotopologue profiling and oxygen consumption experiments of a wild-type (wt) strain and its isogenic mutant. We show that a mutant has significantly lower respiration rates when serine, alanine, pyruvate, α-ketoglutarate or palmitate is the sole carbon source. By contrast, when grown in glucose or glycerol, no differences in respiration were detected. Isotopologue profiling uncovered that the transfer of label from [U-C]serine via pyruvate into the citrate cycle and was lower in the mutant as judged from the labelling patterns of protein-derived amino acids, cell-wall-derived diaminopimelate, sugars and amino sugars and 3-hydroxybutyrate derived from polyhydroxybutyrate (PHB). Similarly, the incorporation of [U-C]glucose via the glycolysis/Entner-Doudoroff (ED) pathway but not via the pentose phosphate pathway was repressed in the mutant. On the other hand, fluxes due to [U-C]glycerol utilization were increased in the mutant. In addition, we showed that exogenous [1,2,3,4-C] is efficiently used for PHB synthesis via C-acetyl-CoA. Taken together, CsrA induces serine catabolism via the tricarboxylic cycle and glucose degradation via the ED pathway, but represses glycerol metabolism, fatty degradation and PHB biosynthesis, in particular during exponential growth. Thus, CsrA has a determining role in substrate usage and carbon partitioning during the life cycle and regulates a switch from amino usage in replicative phase to glycerolipid usage during transmissive growth.© 2017 The Authors.

Keyword: gluconeogenesis

Taurine improves low-level inorganic arsenic-induced insulin resistance by activating PPARγ-mTORC2 signalling and inhibiting hepatic autophagy.

Inorganic arsenic (iAs) is reportedly associated with the increased incidence of type 2 diabetes in the population. Here, we found that iAs exposure significantly decreased the expression of glycolytic genes and glycogen content and increased gene levels in C57/BL6J mice. The expression of peroxisome proliferator-activated receptor γ (PPARγ), and mechanistic target of rapamycin complex 2 (mTORC2) were decreased in the livers of iAs-treated mice. Furthermore, in iAs-treated HepG2 cells, we found that PPARγ agonist rosiglitazone (RGS) increased the expression of mTORC2, inhibited autophagy, and improved glucose metabolism. mTORC2 agonist inhibited autophagy and improved glucose metabolism as well as the autophagosome formation inhibitor 3-methyladenine. Taurine, a natural compound, reversed impaired glucose metabolism and decreased expression of PPARγ and mTORC2 induced by iAs in mice liver and HepG2 cells. These data indicated that taurine administration could ameliorate iAs-induced insulin resistance through activating PPARγ-mTORC2 signalling and subsequently inhibiting hepatic autophagy.© 2018 Wiley Periodicals, Inc.

Keyword: gluconeogenesis

induces central leptin resistance and impairs hepatic glucose and lipid metabolism in male mice.

The consumption of diets rich in saturated fat largely contributes to the development of obesity in modern societies. A diet high in saturated fats can induce inflammation and impair leptin signaling in the hypothalamus. However, the role of saturated fatty acids on hypothalamic leptin signaling, and hepatic glucose and lipid metabolism remains largely undiscovered. In this study, we investigated the effects of intracerebroventricular (icv) administration of a saturated fatty , (PA, C16:0), on central leptin sensitivity, hypothalamic leptin signaling, inflammatory molecules and hepatic energy metabolism in C57BL/6J male mice. We found that the icv administration of PA led to central leptin resistance, evidenced by the inhibition of central leptin's suppression of food intake. Central leptin resistance was concomitant with impaired hypothalamic leptin signaling (JAK2-STAT3, PKB/Akt-FOXO1) and a pro-inflammatory response (TNF-α, IL1-β, IL-6 and pIκBa) in the mediobasal hypothalamus and paraventricular hypothalamic nuclei. Furthermore, the pre-administration of icv PA blunted the effect of leptin-induced decreases in mRNA expression related to (G6Pase and PEPCK), glucose transportation (GLUT2) and lipogenesis (FAS and SCD1) in the liver of mice. Therefore, elevated central PA concentrations can induce pro-inflammatory responses and leptin resistance, which are associated with disorders of energy homeostasis in the liver as a result of diet-induced obesity.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: gluconeogenesis

PCBP2 regulates hepatic insulin sensitivity via HIF-1α and STAT3 pathway in HepG2 cells.

Elevated free fatty acids (FFAs) are fundamental to the pathogenesis of hepatic insulin resistance. However, the molecular mechanisms of insulin resistance remain not completely understood. Transcriptional dysregulation, post-transcriptional modifications and protein degradation contribute to the pathogenesis of insulin resistance. Poly(C) binding proteins (PCBPs) are RNA-binding proteins that are involved in post-transcriptional control pathways. However, there are little studies about the roles of PCBPs in insulin resistance. PCBP2 is the member of the RNA-binding proteins and is thought to participate in regulating hypoxia inducible factor-1 (HIF-1α) and signal transducers and activators of transcription (STAT) pathway which are involved in regulating insulin signaling pathway. Here, we investigated the influence of PCBP2 on hepatic insulin resistance. We showed that the protein and mRNA levels of PCBP2 were down-regulated under insulin-resistant conditions. In addition, we showed that over-expression of PCBP2 ameliorates palmitate (PA)-induced insulin resistance, which was indicated by elevated phosphorylation of protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β). We also found that over-expression of PCBP2 inhibits HIF1α and STAT3 pathway. Furthermore, glucose uptake was found to display a similar tendency with the phosphorylation of Akt. The expressions of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase), two key gluconeogenic enzymes, were down-regulated following Over-expression of PCBP2. Accordingly, PA-induced intracellular lipid accumulation was suppressed in over-expression of PCBP2 HepG2 cells. In addition, we found that over-expression of PCBP2 inhibits HIF1α and STAT3 pathway. Our results demonstrate that PCBP2 was involved in hepatic insulin sensitivity might via HIF-1α and STAT3 pathway in HepG2 cells.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: gluconeogenesis

Pterostilbene reverses mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 diabetes and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: gluconeogenesis

Dipeptidyl peptidase 9 enzymatic activity influences the expression of neonatal metabolic genes.

The success of dipeptidyl peptidase 4 (DPP4) inhibition as a type 2 diabetes therapy has encouraged deeper examination of the post-proline DPP enzymes. DPP9 has been implicated in immunoregulation, disease pathogenesis and metabolism. The DPP9 enzyme-inactive (Dpp9 gene knock-in; Dpp9 gki) mouse displays neonatal lethality, suggesting that DPP9 enzyme activity is essential in neonatal development. Here we present gene expression patterns in these Dpp9 gki neonatal mice. Taqman PCR arrays and sequential qPCR assays on neonatal liver and gut revealed differential expression of genes involved in cell growth, innate immunity and metabolic pathways including long-chain-fatty- uptake and esterification, long-chain fatty acyl-CoA binding, trafficking and transport into mitochondria, lipoprotein metabolism, adipokine transport and in the Dpp9 gki mice compared to wild type. In a liver cell line, Dpp9 knockdown increased AMP-activated protein kinase phosphorylation, which suggests a potential mechanism. DPP9 protein levels in liver cells were altered by treatment with EGF, HGF, insulin or palmitate, suggesting potential natural DPP9 regulators. These gene expression analyses of a mouse strain deficient in DPP9 enzyme activity show, for the first time, that DPP9 enzyme activity regulates metabolic pathways in neonatal liver and gut.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: gluconeogenesis

miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 diabetes mellitus through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 diabetes mellitus (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid metabolism were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of lipid droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and lipid levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic lipid droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid metabolism dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid metabolism in T2DM, which can inhibit hepatic lipogenesis and and elevate glycogen synthesis by targeting STAT3.

Keyword: gluconeogenesis

Resveratrol affects hepatic via histone deacetylase 4.

The aim of this study was to determine whether resveratrol (Rev) affects the expression, phosphorylation, and nuclear and cytoplasmic distribution of histone deacetylase 4 (HDAC4), which in turn affects in hepatocytes under an insulin-resistant state.HepG2 cells were treated with 0.25 mmol/L (PA) to establish an insulin resistance model. The cells were divided into five groups: control, PA, PA + Rev 100 µM, PA + Rev 50 µM, and PA + Rev 20 µM. After treatment for 24 hours, mRNA and protein expression levels of pathway-related molecules and HDAC4 were examined. Next, HepG2 cells were transfected with siRNA-HDAC4. The cells were divided into control, PA, PA + Rev 20 µM, PA + Rev 20 µM +siRNA-HDAC4 negative control, and PA + Rev 20 µM +siRNA-HDAC4 knockdown groups to determine the expression of pathway proteins.Compared with the control group, the pathway-related molecules, glucose-6-phosphatase catalytic subunit (G6PC), phosphoenolpyruvate carboxykinase 1 (PCK1) and forkhead box protein O1 (FOXO1), were increased, and the phosphorylation of FOXO1 decreased after PA treatment. The p-HDAC4 level decreased with the increase in HDAC4 in the nucleus and the decrease in HDAC4 in the cytoplasm in the PA group. Treatment with Rev 20 µM suppressed and promoted HDAC4 shuttling into the cytoplasm from the nucleus. However, 100 and Rev 50 µM exerted the opposite effects. Finally, after HDAC4 knockdown, the expression levels of the key molecules, G6PC, PCK1, and FOXO1, were increased, and p-FOXO1 was decreased, indicating that was enhanced.A low concentration of Rev inhibited under insulin-resistance conditions via translocation of HDAC4 from the nucleus to the cytoplasm.

Keyword: gluconeogenesis

Vernonia amygdalina Delile extract inhibits the hepatic through the activation of adenosine-5'monophosph kinase.

It has been reported that Vernonia amygdalina Delile(VA) presents an anti-diabetic effect, and the effect of VA on lowering glucose is formulated via suppressing the expression of the key hepatic enzyme. Therefore, we further explored the probable mechanism of VA on dismissing hepatic through the activation of adenosine-5' monophosphate kinase (AMPK) in vivo and in vitro.We developed type 2 diabetic mice with STZ and oral administration with VA (50\u202fmg/kg, 100\u202fmg/kg and 150\u202fmg/kg) once a day for 6 weeks. Fasting blood glucose (FBG), fasting insulin (FINS) and oral glucose tolerance tests (OGTT) were conducted. The expression levels of AMPK, phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase) proteins in live were evaluated by western blot. Then, we further explored the mechanism of VA on hepatic in vitro experiments. Glucose production and the expression of AMPK, PEPCK and G6Pase proteins were detected after VA treatment with the presence of the AMPK inhibitor Compound C.VA reduced FBG and caused a significant improvement in glucose tolerance and insulin resistance (HOMA-IR) in STZ-induced mice. VA inhibited the elevated expression of key enzymes (PEPCK and G6Pase) and up-regulated AMPK activity in liver. In (PA)-induced HepG2 cells, VA decreased glucose production and the expression of PEPCK and G6Pase proteins, also activated AMPK pathway. The effects of VA on could be reversed by Compound C.These results reveal that VA suppresses hepatic at least partially through activating the AMPK.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: gluconeogenesis

d- chiro-Inositol Ameliorates High Fat Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic in mice fed a high fat diet and saturated -treated hepatocytes. DCI attenuated free fatty uptake by the liver via lipid trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic . Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic , reduces lipid deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: gluconeogenesis

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: gluconeogenesis

Uncovering sperm metabolome to discover biomarkers for bull fertility.

Subfertility decreases the efficiency of the cattle industry because artificial insemination employs spermatozoa from a single bull to inseminate thousands of cows. Variation in bull fertility has been demonstrated even among those animals exhibiting normal sperm numbers, motility, and morphology. Despite advances in research, molecular and cellular mechanisms underlying the causes of low fertility in some bulls have not been fully elucidated. In this study, we investigated the metabolic profile of bull spermatozoa using non-targeted metabolomics. Statistical analysis and bioinformatic tools were employed to evaluate the metabolic profiles high and low fertility groups. Metabolic pathways associated with the sperm metabolome were also reported.A total of 22 distinct metabolites were detected in spermatozoa from bulls with high fertility (HF) or low fertility (LF) phenotype. The major metabolite classes of bovine sperm were organic acids/derivatives and fatty acids/conjugates. We demonstrated that the abundance ratios of five sperm metabolites were statistically different between HF and LF groups including gamma-aminobutyric (GABA), carbamate, benzoic , lactic , and . Metabolites with different abundances in HF and LF bulls had also VIP scores of greater than 1.5 and AUC- ROC curves of more than 80%. In addition, four metabolic pathways associated with differential metabolites namely alanine, aspartate and glutamate metabolism, β-alanine metabolism, glycolysis or , and pyruvate metabolism were also explored.This is the first study aimed at ascertaining the metabolome of spermatozoa from bulls with different fertility phenotype using gas chromatography-mass spectrometry. We identified five metabolites in the two groups of sires and such molecules can be used, in the future, as key indicators of bull fertility.

Keyword: gluconeogenesis

The BACE1 product sAPPβ induces ER stress and inflammation and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer's disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and fatty oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, insulin resistance, PGC-1α down-regulation, and fatty oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and insulin resistance in skeletal muscle and in liver.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: gluconeogenesis

Exercise performed immediately after fructose ingestion enhances fructose oxidation and suppresses fructose storage.

Exercise prevents the adverse effects of a high-fructose diet through mechanisms that remain unknown.We assessed the hypothesis that exercise prevents fructose-induced increases in very-low-density lipoprotein (VLDL) triglycerides by decreasing the fructose conversion into glucose and VLDL-triglyceride and fructose carbon storage into hepatic glycogen and lipids.Eight healthy men were studied on 3 occasions after 4 d consuming a weight-maintenance, high-fructose diet. On the fifth day, the men ingested an oral (13)C-labeled fructose load (0.75 g/kg), and their total fructose oxidation ((13)CO2 production), fructose storage (fructose ingestion minus (13)C-fructose oxidation), fructose conversion into blood (13)C glucose ( from fructose), blood VLDL-(13)C palmitate (a marker of hepatic de novo lipogenesis), and lactate concentrations were monitored over 7 postprandial h. On one occasion, participants remained lying down throughout the experiment [fructose treatment alone with no exercise condition (NoEx)], and on the other 2 occasions, they performed a 60-min exercise either 75 min before fructose ingestion [exercise, then fructose condition (ExFru)] or 90 min after fructose ingestion [fructose, then exercise condition (FruEx)].Fructose oxidation was significantly (P < 0.001) higher in the FruEx (80% ± 3% of ingested fructose) than in the ExFru (46% ± 1%) and NoEx (49% ± 1%). Consequently, fructose storage was lower in the FruEx than in the other 2 conditions (P < 0.001). Fructose conversion into blood (13)C glucose, VLDL-(13)C palmitate, and postprandial plasma lactate concentrations was not significantly different between conditions.Compared with sedentary conditions, exercise performed immediately after fructose ingestion increases fructose oxidation and decreases fructose storage. In contrast, exercise performed before fructose ingestion does not significantly alter fructose oxidation and storage. In both conditions, exercise did not abolish fructose conversion into glucose or its incorporation into VLDL triglycerides. This trial was registered at clinicaltrials.gov as .© 2016 American Society for Nutrition.

Keyword: gluconeogenesis

lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress by targeting activating transcription factor 4 (ATF4), which regulates by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑fat diet (HFD)‑fed mice to verify the role of MEG3 in hepatic insulin resistance in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 diabetes mellitus.

Keyword: gluconeogenesis

A novel PPARα/γ agonist, propane-2-sulfonic octadec-9-enyl-amide, ameliorates insulin resistance and in vivo and vitro.

Peroxisome proliferator-activated receptor alpha/gamma (PPARα/γ) agonists have emerged as important pharmacological agents for improving insulin action. Propane-2-sulfonic octadec-9-enyl-amide (N15) is a novel PPARα/γ dual agonist synthesized in our laboratory. The present study investigates the efficacy and safety of N15 on insulin resistance regulation in high fat diet (HFD)-and streptozotocin (STZ)-induced diabetic mice and in (PA)-induced HepG2 cells. Our results showed that N15 remarkably ameliorated insulin resistance and dyslipidemia in vivo, as well as rectified the glucose consumption and in vitro. Moreover, the glucose-lowering effect of N15 was associated with PPARγ mediated up-regulation of hepatic glucose consumption and down-regulation of . Meanwhile, N15 exerted advantageous effects on glucose and lipid metabolism without triggering weight gain and hepatotoxicity in mice. In conclusion, our data demonstrated that by alleviating glucose and lipid abnormalities, N15 could be used as a potential prophylactic and therapeutic agent against type 2 diabetes and related metabolic disorders.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: gluconeogenesis

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: gluconeogenesis

Flavin-Containing Monooxygenase 3 Reduces Endoplasmic Reticulum Stress in Lipid-Treated Hepatocytes.

Isoforms of flavin-containing monooxygenase (FMO) are involved in xenobiotic metabolism but have also been implicated in the regulation of glucose and lipid homeostasis and in the development of atherosclerosis. However, we have recently shown that improved insulin action is associated with increased FMO expression in livers of protein kinase C-deficient mice. Here, we investigated whether FMO3 expression affected insulin signaling, glucose metabolism, and endoplasmic reticulum (ER) stress in hepatocytes. HepG2 and IHH hepatocytes were transfected with FMO3 cDNA for overexpression, or small interfering RNA for knockdown. Cells were treated with palmitate to induce insulin resistance and insulin signaling, phosphoenolpyruvate carboxykinase (PEPCK) gene expression and ER stress markers were examined by immunoblotting and RT-PCR. Glycogen synthesis was measured using [(14)C]glucose. Palmitate treatment reduced insulin signaling at the level of Akt phosphorylation and glycogen synthesis, which were little affected by FMO3 overexpression. However, the fatty also increased the levels of several ER stress markers and activation of caspase 3, which were counteracted by FMO3 overexpression and exacerbated by FMO3 knockdown. Although FMO3 expression did not reverse lipid effects on protein thiol redox in hepatocytes, it did prevent up-regulation of the gluconeogenic enzyme PEPCK by pharmacological ER stress inducers or by palmitate. ER stress and PEPCK levels were also reduced in livers of fat-fed protein kinase Cδ-deficient mice. Our data indicate that FMO3 can contribute to the regulation of glucose metabolism in the liver by reducing lipid-induced ER stress and the expression of PEPCK, independently of insulin signal transduction.

Keyword: gluconeogenesis

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Alteration of glycogen and glucose metabolism in ischaemic and post-ischaemic working rat hearts by adenosine A1 receptor stimulation.

1. Cardioprotection by adenosine A1 receptor activation limits infarct size and improves post-ischaemic mechanical function. The mechanisms responsible are unclear but may involve alterations in myocardial glucose metabolism. 2. Since glycogen is an important source of glucose during ischaemia, we examined the effects of N6-cyclohexyladenosine (CHA), an A1 receptor agonist, on glycogen and glucose metabolism during ischaemia as well as reperfusion. 3. Isolated working rat hearts were perfused with Krebs-Henseleit solution containing dual-labelled 5-3H and 14C glucose and palmitate as energy substrates. Rates of and glucose oxidation were measured directly from the production of 3H2O and 14CO2. Glycogen turnover was measured from the rate of change of [5-3H and 14C]glucosyl units in total myocardial glycogen. 4. Following low-flow (0.5 ml min-1) ischaemia (60 min) and reperfusion (30 min), left ventricular minute work (LV work) recovered to 22% of pre-ischaemic values. CHA (0.5 microM) improved the recovery of LV work 2 fold. 5. CHA altered glycogen turnover in post-ischaemic hearts by stimulating glycogen synthesis while having no effects on glycogen degradation. CHA also partially inhibited . These changes accelerated the recovery of glycogen in CHA-treated hearts and reduced proton production. 6. During ischaemia, CHA had no measurable effect on glycogen turnover or glucose metabolism. Glycogen phosphorylase activity, which was elevated after ischaemia, was inhibited by CHA, possibly in response to CHA-induced inhibition of AMP-activated protein kinase activity. 7. These results indicate that CHA-induced cardioprotection is associated with alterations of glycogen turnover during reperfusion as well as improved metabolic coupling of to glucose oxidation.

Keyword: glycolysis

Flux-balance analysis of mitochondrial energy metabolism: consequences of systemic stoichiometric constraints.

Mitochondrial metabolism is a critical component in the functioning and maintenance of cellular organs. The stoichiometry of biochemical reaction networks imposes constraints on mitochondrial function. A modeling framework, flux-balance analysis (FBA), was used to characterize the optimal flux distributions for maximal ATP production in the mitochondrion. The model predicted the expected ATP yields for glucose, lactate, and palmitate. Genetic defects that affect mitochondrial functions have been implicated in several human diseases. FBA can characterize the metabolic behavior due to genetic deletions at the metabolic level, and the effect of mutations in the tricarboxylic (TCA) cycle on mitochondrial ATP production was simulated. The mitochondrial ATP production is severely affected by TCA-cycle mutations. In addition, the model predicts the secretion of TCA-cycle intermediates, which is observed in clinical studies of mitochondriopathies such as those associated with fumarase deficiency. The model provides a systemic perspective to characterize the effect of stoichiometric constraints and specific metabolic fluxes on mitochondrial function.

Keyword: glycolysis

Fat metabolism during exercise in patients with mitochondrial disease.

To determine whether patients with defects of the respiratory chain have metabolic adaptations that promote a preferential use of fats or carbohydrates, similar to what is observed in metabolic myopathies affecting or fat oxidation.Causation and case-control study. Fat metabolism was determined by means of indirect calorimetry and stable isotope technique in patients and healthy subjects. Patients carried various types and loads (mean [SE], 72% [5%]) of mitochondrial DNA (mtDNA) mutations in skeletal muscle. All subjects exercised at the same absolute workload (mean [SE], 65 [10] W), corresponding to 72% (in patients) and 30% (in healthy subjects) of maximum oxygen consumption.Neuromuscular research unit.Ten patients with mtDNA mutations and 10 sex-matched healthy subjects.Fat turnover, plasma concentrations of palmitate and total free fatty acids, glucose mobilization, and total carbohydrate oxidation.Fat turnover and plasma concentrations of palmitate and total free fatty acids were similar in patients and healthy subjects at rest and during exercise. In line with the higher relative workload of the patients, glucose mobilization and total carbohydrate oxidation were higher in the patients compared with the healthy subjects.During moderate-intensity exercise, the balance between fat and carbohydrate use in patients with mtDNA mutations matches that seen in healthy subjects, indicating that manipulating dietary fat and carbohydrate content is not a feasible therapeutic option to improve exercise intolerance in these disorders.

Keyword: glycolysis

Epinephrine increases ATP production in hearts by preferentially increasing glucose metabolism.

Although epinephrine is widely used clinically, its effect on myocardial energy substrate preference in the intact heart has yet to be clearly defined. We determined the effects of epinephrine on glucose and fatty metabolism in isolated working rat hearts perfused with 11 mM glucose, 0.4 mM palmitate, and 100 muU/ml insulin at an 11.5-mmHg left atrial preload and a 60-mmHg aortic afterload. and glucose oxidation were measured in hearts perfused with [5-3H]glucose and [U-14C]glucose, whereas fatty oxidation was measured in hearts perfused with [1-14C]palmitate. Addition of 1 microM epinephrine resulted in a 53% increase in the heart rate-developed pressure product. increased dramatically following addition of epinephrine (a 272% increase), as did glucose oxidation (a 410% increase). In contrast, fatty oxidation increased by only 10%. Epinephrine treatment did not increase the amount of oxygen required to produce an equivalent amount of ATP; however, epinephrine did increase the uncoupling between and glucose oxidation in these fatty -perfused hearts, resulting in a significant increase in H+ production from glucose metabolism. Overall ATP production in epinephrine-treated hearts increased 59%. The contribution of glucose ( and glucose oxidation) to ATP production increased from 13 to 36%, which was accompanied by a reciprocal decrease in the contribution of fatty oxidation to ATP production from 83 to 63%. The increase in glucose oxidation was accompanied by a significant increase in pyruvate dehydrogenase complex activity in the active form. We conclude that the increase in ATP required for contractile function following epinephrine treatment occurs through a preferential increase in glucose use.

Keyword: glycolysis

Metabolic Profiles Reveal Changes in Wild and Cultivated Soybean Seedling Leaves under Salt Stress.

Clarification of the metabolic mechanisms underlying salt stress responses in plants will allow further optimization of crop breeding and cultivation to obtain high yields in saline-alkali land. Here, we characterized 68 differential metabolites of cultivated soybean (Glycine max) and wild soybean (Glycine soja) under neutral-salt and alkali-salt stresses using gas chromatography-mass spectrometry (GC-MS)-based metabolomics, to reveal the physiological and molecular differences in salt tolerance. According to comparisons of growth parameters under the two kinds of salt stresses, the level of inhibition in wild soybean was lower than in cultivated soybean, especially under alkali-salt stress. Moreover, wild soybean contained significantly higher amounts of phenylalanine, asparagine, citraconic , citramalic , citric and α-ketoglutaric under neutral-salt stress, and higher amounts of , lignoceric , glucose, citric and α-ketoglutaric under alkali-salt stress, than cultivated soybean. Further investigations demonstrated that the ability of wild soybean to salt tolerance was mainly based on the synthesis of organic and amino acids, and the more active tricarboxylic cycle under neutral-salt stress. In addition, the metabolite profiling analysis suggested that the generation from β-oxidation, glycolysis and the citric cycle plays important roles under alkali-salt stress. Our results extend the understanding of mechanisms involved in wild soybean salt tolerance and provide an important reference for increasing yields and developing salt-tolerant soybean cultivars.

Keyword: glycolysis

Hemin increases aerobic capacity of cultured regenerating skeletal myotubes.

Regeneration of damaged, mature muscle occurs by differentiation of satellite cells. In culture, satellite cell myoblasts proliferate, align, and fuse to form cross-striated, contracting myotubes. The biochemical changes and the factors that regulate differentiation in satellite cells have not been investigated previously. We report here that no significant differences in glucose uptake rate or glucose oxidation rate were observed between regenerating myoblasts and myotubes, whereas the aerobic oxidation of increased 7.3-fold between these differentiation states. Specific activities of enzymes of critical importance in aerobic metabolism or in production of ATP were increased 2- to 3.5-fold during fusion. Addition of 20 microM hemin to regenerating muscle cultures potentiated the aerobic capacity as evidenced by a 23.6% increase in palmitate oxidation rate. Hemin also increased the specific activities of all nonheme enzymes investigated with the exception of phosphofructokinase. This augmentation of aerobic metabolism together with the time frame of active muscle differentiation suggests a complex role for hemin in myogenesis.

Keyword: glycolysis

CsrA regulates a metabolic switch from amino to glycerolipid metabolism.

CsrA plays a crucial role in the life-stage-specific expression of virulence phenotypes and metabolic activity. However, its exact role is only partly known. To elucidate how CsrA impacts metabolism we analysed the CsrA depended regulation of metabolic functions by comparative C-isotopologue profiling and oxygen consumption experiments of a wild-type (wt) strain and its isogenic mutant. We show that a mutant has significantly lower respiration rates when serine, alanine, pyruvate, α-ketoglutarate or palmitate is the sole carbon source. By contrast, when grown in glucose or glycerol, no differences in respiration were detected. Isotopologue profiling uncovered that the transfer of label from [U-C]serine via pyruvate into the citrate cycle and gluconeogenesis was lower in the mutant as judged from the labelling patterns of protein-derived amino acids, cell-wall-derived diaminopimelate, sugars and amino sugars and 3-hydroxybutyrate derived from polyhydroxybutyrate (PHB). Similarly, the incorporation of [U-C]glucose via the /Entner-Doudoroff (ED) pathway but not via the pentose phosphate pathway was repressed in the mutant. On the other hand, fluxes due to [U-C]glycerol utilization were increased in the mutant. In addition, we showed that exogenous [1,2,3,4-C] is efficiently used for PHB synthesis via C-acetyl-CoA. Taken together, CsrA induces serine catabolism via the tricarboxylic cycle and glucose degradation via the ED pathway, but represses glycerol metabolism, fatty degradation and PHB biosynthesis, in particular during exponential growth. Thus, CsrA has a determining role in substrate usage and carbon partitioning during the life cycle and regulates a switch from amino usage in replicative phase to glycerolipid usage during transmissive growth.© 2017 The Authors.

Keyword: glycolysis

Adiponectin induces insulin secretion in vitro and in vivo at a low glucose concentration.

A decrease in plasma adiponectin levels has been shown to contribute to the development of diabetes. However, it remains uncertain whether adiponectin plays a role in the regulation of insulin secretion. In this study, we investigated whether adiponectin may be involved in the regulation of insulin secretion in vivo and in vitro.The effect of adiponectin on insulin secretion was measured in vitro and in vivo, along with the effects of adiponectin on ATP generation, membrane potentials, Ca2+ currents, cytosolic calcium concentration and state of 5\'-AMP-activated protein kinase (AMPK). In addition, insulin granule transport was measured by membrane capacitance and total internal reflection fluorescence (TIRF) analysis.Adiponectin significantly stimulated insulin secretion from pancreatic islets to approximately 2.3-fold the baseline value in the presence of a glucose concentration of 5.6 mmol/l. Although adiponectin had no effect on ATP generation, membrane potentials, Ca2+ currents, cytosolic calcium concentrations or activation status of AMPK, it caused a significant increase of membrane capacitance to approximately 2.3-fold the baseline value. TIRF analysis revealed that adiponectin induced a significant increase in the number of fusion events in mouse pancreatic beta cells under 5.6 mmol/l glucose loading, without affecting the status of previously docked granules. Moreover, intravenous injection of adiponectin significantly increased insulin secretion to approximately 1.6-fold of baseline in C57BL/6 mice.The above results indicate that adiponectin induces insulin secretion in vitro and in vivo.

Keyword: glycolysis

Multi-omics analysis points to altered platelet functions in severe food-associated respiratory allergy.

Prevalence and severity of allergic diseases have increased worldwide. To date, respiratory allergy phenotypes are not fully characterized and, along with inflammation progression, treatment is increasingly complex and expensive. Profilin sensitization constitutes a good model to study the progression of allergic inflammation. Our aim was to identify the underlying mechanisms and the associated biomarkers of this progression, focusing on severe phenotypes, using transcriptomics and metabolomics.Twenty-five subjects were included in the study. Plasma samples were analyzed using gas and liquid chromatography coupled to mass spectrometry (GC-MS and LC-MS, respectively). Individuals were classified in four groups-"nonallergic," "mild," "moderate," and "severe"-based on their clinical history, their response to an oral challenge test with profilin, and after a refinement using a mathematical metabolomic model. PBMCs were used for microarray analysis.We found a set of transcripts and metabolites that were specific for the "severe" phenotype. By metabolomics, a decrease in carbohydrates and pyruvate and an increase in lactate were detected, suggesting aerobic . Other metabolites were incremented in "severe" group: lysophospholipids, sphingosine-1-phosphate, sphinganine-1-phosphate, and lauric, myristic, , and oleic fatty acids. On the other hand, carnitines were decreased along severity. Significant transcripts in the "severe" group were found to be downregulated and were associated with platelet functions, protein synthesis, histone modification, and fatty metabolism.We have found evidence that points to the association of severe allergic inflammation with platelet functions alteration, together with reduced protein synthesis, and switch of immune cells to aerobic .© 2018 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

Keyword: glycolysis

Metabolic regulation of NLRP3.

A shift in our understanding of macrophage biology has come about as a result of recent discoveries in the area of metabolic reprogramming of macrophages. The NLRP3 inflammasome drives the activation of caspase-1, leading to the production of IL-1β, IL-18, and a type of cell death termed pyroptosis. The NLRP3 inflammasome has been shown to sense metabolites such as palmitate, uric , and cholesterol crystals and is inhibited by ketone bodies produced during metabolic flux. The NLRP3 inflammasome has also been shown to be regulated by mitochondrial reactive oxygen species and components of , such as Hexokinase. Here, we review these findings and discuss their importance for inflammation and furthermore discuss potential therapeutic benefits of targeting NLRP3.© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: glycolysis

Effects of high levels of fatty acids on functional recovery of ischemic hearts from diabetic rats.

In this study we determined the effects of high levels of fatty acids on recovery of heart function when present either during or after ischemia. Isolated working hearts from 6-wk streptozotocin diabetic and control rats perfused with 11 mM glucose were subjected to 25 min of global ischemia followed by 30 min of aerobic reperfusion. Four groups were studied: 1) 1.2 mM palmitate present before, during, and after ischemia; 2) 1.2 mM palmitate present before and during ischemia, followed by reperfusion in the absence of palmitate; 3) no palmitate before and during ischemia, followed by 1.2 mM palmitate during reperfusion; and 4) no palmitate before and during ischemia or during reperfusion. In control hearts, palmitate during reperfusion depressed recovery of function regardless of whether palmitate was present or absent during ischemia. In contrast, palmitate present during reperfusion did not depress recovery of mechanical function in the diabetic rat hearts. However, the presence of palmitate during ischemia itself in diabetic rat hearts was detrimental to recovery of mechanical function. The presence of palmitate during ischemia resulted in an accelerated rate of ATP loss and a decreased rate of lactate accumulation during ischemia, although this effect was similar in control and diabetic rat hearts. Our results demonstrate that high concentrations of fatty acids depress functional recovery of control rat hearts during the reperfusion period but depress recovery of function in diabetic rat hearts when present during ischemia itself.

Keyword: glycolysis

Epinephrine plasma thresholds for lipolytic effects in man: measurements of fatty transport with [l-13C].

To determine the plasma epinephrine thresholds for its lipolytic effect, 60-min epinephrine infusions at nominal rates of 0.1, 0.5, 1.0, 2.5, and 5.0 micrograms/min were performed in each of four normal young adult men while they also received a simultaneous infusion of [1-13C] to estimate inflow transport of plasma free fatty acids. These 20 infusions resulted in steady-state plasma epinephrine concentrations ranging from 12 to 870 pg/ml. Plasma epinephrine thresholds for changes in blood glucose, lactate, and beta-hydroxybutyrate were in the 150--200-pg/ml range reported by us previously (Clutter, W. E., D. M. Bier, S. D. Shah, and P. E. Cryer. 1980. J. Clin. Invest. 66: 94--101.). Increments in plasma glycerol and free fatty acids and in the inflow and outflow transport of palmitate, however, occurred at lower plasma epinephrine thresholds in the range of 75 to 125 pg/ml. Palmitate clearance was unaffected at any steady-state epinephrine level produced. These data indicate that (a) the lipolytic effects of epinephrine occur at plasma levels approximately threefold basal values and (b) lipolysis is more sensitive than glycogenolysis to increments in plasma epinephrine.

Keyword: glycolysis

The AMPK gamma1 R70Q mutant regulates multiple metabolic and growth pathways in neonatal cardiac myocytes.

Although mutations in the gamma-subunit of AMP-activated protein kinase (AMPK) can result in excessive glycogen accumulation and cardiac hypertrophy, the mechanisms by which this occurs have not been well defined. Because >65% of cardiac AMPK activity is associated with the gamma1-subunit of AMPK, we investigated the effects of expression of an AMPK-activating gamma1-subunit mutant (gamma1 R70Q) on regulatory pathways controlling glycogen accumulation and cardiac hypertrophy in neonatal rat cardiac myocytes. Whereas expression of gamma1 R70Q displayed the expected increase in palmitate oxidation rates, rates of were significantly depressed. In addition, glycogen synthase activity was increased in cardiac myocytes expressing gamma1 R70Q, due to both increased expression and decreased phosphorylation of glycogen synthase. The inhibition of and increased glycogen synthase activity were correlated with elevated glycogen levels in gamma1 R70Q-expressing myocytes. In association with the reduced phosphorylation of glycogen synthase, glycogen synthase kinase (GSK)-3beta protein and mRNA levels were profoundly decreased in the gamma1 R70Q-expressing myocytes. Consistent with GSK-3beta negatively regulating hypertrophy via inhibition of nuclear factor of activated T cells (NFAT), the dramatic downregulation of GSK-3beta was associated with increased nuclear activity of NFAT. Together, these data provide important new information about the mechanisms by which a mutation in the gamma-subunit of AMPK causes altered AMPK signaling and identify multiple pathways involved in regulating both cardiac myocyte metabolism and growth that may contribute to the development of the gamma mutant-associated cardiomyopathy.

Keyword: glycolysis

Time-course metabolic profiling in alfalfa leaves under Phoma medicaginis infection.

Information on disease process and pathogenicity mechanisms is important for understanding plant disease. Spring black stem and leaf spot caused by the necrotrophic pathogen Phoma medicaginis var. medicaginis Malbr. & Roum causes large losses to alfalfa. However, till now, little is known about alfalfa-P. medicagnis interactions and the pathogenicity mechanisms of the pathogen. Here, alfalfa inoculated with P. medicaginis was subjected to GC-MS based metabolic profiling. The metabolic response in P. medicaginis-inoculated and mock-inoculated alfalfa leaves was assessed at 2, 4, 6, 8, 12, 16, 20, 24, 26 and 28 days post inoculation. In total, 101 peaks were detected in the control and inoculated groups, from which 70 metabolites were tentatively identified. Using multivariate analysis, 16 significantly regulated compounds, including amino acids, nitrogen-containing compounds and organic acids, polyols, fatty acids, and sugars were tentatively identified (Variable importance values, VIP>1.0 and p <0.05). Among these metabolites, the levels of malate, 5-oxoproline, and stearic were increased significantly in P. medicaginis-infected alfalfa leaves compared to the controls. In contrast, the levels ofγ-aminobutyric and 2-pyrrolidinone were significantly decreased in infected leaves compared to the controls. Further metabolic pathway analysis of the 16 significantly regulated compounds demonstrated that , the tricarboxylic cycle, and β-oxidation of fatty acids were significantly induced in the alfalfa leaves at later stages of P. medicaginis infection. The strong induction of tricarboxylic cycle pathways at later infection stages caused by the pathogen may induce senescence in the alfalfa leaves, leading to plant death. However, intermediate metabolites of these metabolic pathways, and inositol phosphate, glutathione, the metabolic pathways of some amino acids accumulated rapidly and strongly at early stages of infection, which may enhance the ability of alfalfa to resist necrotrophic P. medicaginis disease. Understanding metabolic pathways is essential for understanding pathogenesis.

Keyword: glycolysis

Second window of preconditioning normalizes palmitate use for oxidation and improves function during low-flow ischaemia.

Although a major mechanism for cardioprotection is altered metabolism, little is known regarding metabolic changes in ischaemic preconditioning and subsequent ischaemia. Our objective was to examine the effects of the second window of preconditioning (SWOP), the delayed phase of preconditioning against infarction and stunning, on long-chain free fatty (LCFA) oxidation during ischaemia in chronically instrumented, conscious pigs.We studied three groups: (i) normal baseline perfusion (n = 5); (ii) coronary artery stenosis (CAS; n = 5); (iii) CAS 24 h following 2 × 10 min coronary occlusions and 10 min reperfusion (n = 7). Ischaemia was induced by a left anterior descending (LAD) stenosis (40% flow reduction) for 90 min, dropping systolic wall thickening by 72%. LCFA oxidation was assessed following LAD infusion of (13)C palmitate, i.e. during control or stenosis, by in vitro nuclear magnetic resonance of the sampled myocardium. Stenosis reduced subendocardial blood flow subendocardially, but not subepicardial, yet induced transmural reductions in LCFA oxidation and increased non-oxidative . During stenosis, preconditioned hearts showed normalized contributions of LCFA to oxidative ATP synthesis, despite increased lactate accumulation. SWOP induced a shift towards LCFA oxidation during stenosis, despite increased malonyl-CoA, and marked protection of contractile function with a significant improvement in systolic wall thickening.Thus, the second window of preconditioning normalized oxidative metabolism of LCFA during subsequent ischaemia despite elevated non-oxidative and malonyl-CoA and was linked to protection of regional contractile function resulting in improved mechanical performance. Interestingly, the metabolic responses occurred transmurally while ischaemia was restricted solely to the subendocardium.

Keyword: glycolysis

Contribution of oxidative metabolism and to ATP production in hypertrophied hearts.

The contribution of and oxidative metabolism to ATP production was determined in isolated working hypertrophied hearts perfused with Krebs-Henseleit buffer containing 3% albumin, 0.4 mM palmitate, 0.5 mM lactate, and 11 mM glucose. and glucose oxidation were directly measured by perfusing hearts with [5-3H/U-14C]glucose and by measuring 3H2O and 14CO2 production, respectively. Palmitate and lactate oxidation were determined by simultaneous measurement of 3H2O and 14CO2 in hearts perfused with [9,10-3H]palmitate and [U-14C]lactate. At low workloads (60 mmHg aortic after-load), rates of palmitate oxidation were 47% lower in hypertrophied hearts than in control hearts, but palmitate oxidation remained the primary energy source in both groups, accounting for 55 and 69% of total ATP production, respectively. The contribution of to ATP production was significantly higher in hypertrophied hearts (19%) than in control hearts (7%), whereas that of glucose and lactate oxidation did not differ between groups. During conditions of high work (120 mmHg aortic afterload), the extra ATP production required for mechanical function was obtained primarily from an increase in the oxidation of glucose and lactate in both groups. The contribution of palmitate oxidation to overall ATP production decreased in hypertrophied and control hearts (to 40 and 55% of overall ATP production, respectively) and was no longer significantly depressed in hypertrophied hearts. , on the other hand, was accelerated in control hearts to rates seen in the hypertrophied hearts. Thus a reduced contribution of fatty oxidation to energy production in hypertrophied rat hearts is accompanied by a compensatory increase in during low work conditions.(ABSTRACT TRUNCATED AT 250 WORDS).

Keyword: glycolysis

Effects of anoxia and low free fatty on myocardial energy metabolism in streptozotocin-diabetic rats.

To investigate anaerobic and glucose oxidation in the diabetic heart, we studied the effects of anoxia and low exogenous free fatty (FFA) on myocardial adenosine triphosphate (ATP) content in the isolated perfused heart from streptozotocin-diabetic rats. The perfusate consisted of Krebs-Ringer buffer (pH 7.4) containing 5 mM glucose and 0.30 mM or 0.05 mM FFA, equilibrated with 95% O2 + 5% CO2 or 95% N2 + 5% CO2 gas. Under aerobic conditions, glucose uptake, lactate production and glucose oxidation by the diabetic heart were reduced by 68, 57 and 53% of the control, respectively, and FFA oxidation by the diabetic heart was increased by 67% of the control. Under anoxic conditions, the increments in glucose uptake and lactate production by the diabetic heart were much smaller than those by the control (2.6 and 9.5 mumol/g/30 min vs. 11.4 and 40 mumol/g/30 min, respectively). The ATP content in the diabetic myocardium was decreased more rapidly in the anoxic perfusion than that in the control. In the aerobic perfusion, the ATP content in the control myocardium was maintained for 60 min at low FFA (0.05 mM) in the perfusate. The ATP content in the diabetic myocardium was reduced after 40 min at low FFA but was well maintained for 60 min by the addition of 0.30 mM FFA. Thus the ATP content in aerobic diabetic myocardium was not well maintained when exogenous FFA supply was depleted. These results suggest that diabetic hearts have a primary defect in the stimulation of and glucose oxidation, which does not result from increased FFA utilization.

Keyword: glycolysis

Hydrodynamic and energetic aspects of exogenous free fatty perfusion in the isolated rat heart during high flow anoxia and reoxygenation: a 31P magnetic resonance study.

The aim was to show differences between the effects of various dietary long chain fatty acids (, oleic, linoleic, alpha and gamma linolenic acids) perfused in isolated rat hearts subjected to a sequence of high flow anoxia and subsequent reoxygenation.Isolated working rat hearts perfused with selected exogenous fatty acids were allowed an initial 30 min equilibration period followed by 60 min of high flow anoxia and 40 min of reoxygenation. Ventricular function and tissue contents of phosphorylated metabolites were monitored concomitantly using standard procedures and 31P magnetic resonance spectroscopy respectively.Hearts were removed from male Wistar rats weighing 350-400 g. Results from eight hearts were pooled for each of the five fatty acids perfused.High coronary flow maintained during anoxia led to an increased extracellular washout of lactate and only to mild intracellular acidosis, limiting myocardial damage by metabolites of anaerobic . Under these conditions, marked differences between the classes of perfused fatty acids were observed. Hearts which received oleic showed the most depressed ventricular function and a greater depletion in high energy phosphates content. These deleterious effects were completely reversed by the separate administration of two fatty metabolism blocking agents, nicotinic and oxfenicine. Cardioprotection was enhanced by perfusion of polyunsaturated fatty acids (linoleic , alpha and gamma linolenic acids). Hearts perfused either with glucose or with behaved similarly and showed an intermediate functional and metabolic postanoxic recovery.This study documents the relation between the chemical structure of exogenous fatty acids used in heart perfusion and their ability to improve or impair postanoxic myocardial recovery. The cardioprotective effect of polyunsaturated fatty acids was documented by simultaneous evaluation of mechanical performance and metabolic response.

Keyword: glycolysis

Effect of prolonged starvation on substrate uptake in the isolated perfused rat heart.

The pattern of substrate uptake by the heart in prolonged starvation, when lipid reserves are approaching depletion, has been examined. The classical Langendorff perfused heart preparation was employed to determine substrate uptakes in male rats fed ad libitum or starved for 7 days. Levels of metabolites in "arterial" and "venous" perfusion media and in heart tissue were determined by fluoroenzymatic assays, with the exception of which was analyzed by gas chromatography. It was found that glucose is the principal fuel of oxidation in perfused hearts of ad libitum-fed rats, whereas palmitate (FFA) is the major fuel of oxidation in perfused hearts of starved rats, followed by lactate, glucose, beta-hydroxybutyrate, pyruvate and alanine. Such changes might be related to some of the alterations in the metabolic pathway (e.g., glycolytic inhibition) in prolonged starvation.

Keyword: glycolysis

Pigment epithelium-derived factor (PEDF) regulates metabolism and insulin secretion from a clonal rat pancreatic beta cell line BRIN-BD11 and mouse islets.

Pigment epithelium-derived factor (PEDF) is a multifunctional glycoprotein, associated with lipid catabolism and insulin resistance. In the present study, PEDF increased chronic and acute insulin secretion in a clonal rat β-cell line BRIN-BD11, without alteration of glucose consumption. PEDF also stimulated insulin secretion from primary mouse islets. Seahorse flux analysis demonstrated that PEDF did not change mitochondrial respiration and glycolytic function. The cytosolic presence of the putative PEDF receptor - adipose triglyceride lipase (ATGL) - was identified, and ATGL associated stimulation of glycerol release was robustly enhanced by PEDF, while intracellular ATP levels increased. Addition of palmitate or ex\xa0vivo stimulation with inflammatory mediators induced β-cell dysfunction, effects not altered by the addition of PEDF. In conclusion, PEDF increased insulin secretion in BRIN-BD11 and islet cells, but had no impact on glucose metabolism. Thus elevated lipolysis and enhanced fatty availability may impact insulin secretion following PEDF receptor (ATGL) stimulation.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: glycolysis

Hepatocellular toxicity of benzbromarone: effects on mitochondrial function and structure.

Benzbromarone is an uricosuric structurally related to amiodarone and a known mitochondrial toxicant. The aim of the current study was to improve our understanding in the molecular mechanisms of benzbromarone-associated hepatic mitochondrial toxicity. In HepG2 cells and primary human hepatocytes, ATP levels started to decrease in the presence of 25-50μM benzbromarone for 24-48h, whereas cytotoxicity was observed only at 100μM. In HepG2 cells, benzbromarone decreased the mitochondrial membrane potential starting at 50μM following incubation for 24h. Additionally, in HepG2 cells, 50μM benzbromarone for 24h induced mitochondrial uncoupling,and decreased mitochondrial ATP turnover and maximal respiration. This was accompanied by an increased lactate concentration in the cell culture supernatant, reflecting increased as a compensatory mechanism to maintain cellular ATP. Investigation of the electron transport chain revealed a decreased activity of all relevant enzyme complexes. Furthermore, treatment with benzbromarone was associated with increased cellular ROS production, which could be located specifically to mitochondria. In HepG2 cells and in isolated mouse liver mitochondria, benzbromarone also reduced metabolism due to an inhibition of the long-chain acyl CoA synthetase. In HepG2 cells, benzbromarone disrupted the mitochondrial network, leading to mitochondrial fragmentation and a decreased mitochondrial volume per cell. Cell death occurred by both apoptosis and necrosis. The study demonstrates that benzbromarone not only affects the function of mitochondria in HepG2 cells and human hepatocytes, but is also associated with profound changes in mitochondrial structure which may be associated with apoptosis.Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Keyword: glycolysis

Cardioprotection by GSK-3 inhibition: role of enhanced glycogen synthesis and attenuation of calcium overload.

Glycogen synthase kinase-3 (GSK-3) is a multi-functional kinase that regulates signalling pathways affecting glycogen metabolism, protein synthesis, mitosis, and apoptosis. GSK-3 inhibition limits cardiac ischaemia-reperfusion (IR) injury, but mechanisms are not clearly defined. This study tested the hypothesis that acute GSK-3 inhibition stimulates glycogen synthesis, repartitions glucose away from , reduces proton (H+) production from glucose metabolism, and attenuates intracellular Ca2+ (Ca2+(i)) overload.In isolated perfused working rat hearts subjected to global ischaemia and reperfusion, the selective GSK-3 inhibitor, SB-216763 (SB, 3 micromol/L), when added either prior to ischaemia or at the onset of reperfusion, improved recovery of left-ventricular (LV) work. SB increased glycogen synthesis during reperfusion while and H+ production were reduced. Rates of glucose and palmitate oxidation were improved by SB. Measurement of Ca2+(i) concentration by rapid acquisition indo-1 fluorescence imaging showed that SB, when added either prior to ischaemia or at the onset of reperfusion, reduced diastolic Ca2+(i) overload during reperfusion. In aerobic hearts depleted of glycogen by substrate-free perfusion to a level similar to that measured at the onset of reperfusion, SB accelerated glycogen synthesis and reduced and H+ production independent of changes in LV work.Our study indicates that reduction in H+ production by GSK-3 inhibition is an early and upstream event that lessens Ca2+(i) overload during ischaemia and early reperfusion independent of LV work which enhances the recovery of post-ischaemic LV function and that may ultimately contribute to previously observed reductions in cell death and infarction.

Keyword: glycolysis

Roles and regulation of ketogenesis in cultured astroglia and neurons under hypoxia and hypoglycemia.

Exogenous ketone bodies (KBs), acetoacetate (AA), and β-hydroxybutyrate (BHB) act as alternative energy substrates in neural cells under starvation. The present study examined the endogenous ketogenic capacity of astroglia under hypoxia with/without glucose and the possible roles of KBs in neuronal energy metabolism. Cultured neurons and astroglia were prepared from Sprague-Dawley rats. (PAL) and l-carnitine (LC) were added to the assay medium. The 4- to 24-hr production of AA and BHB was measured using the cyclic thio-NADH method. (14)C-labeled -soluble products (KBs) and (14)CO2 produced from [1-(14)C]PAL were also measured. l-[U-(14)C]lactic ([(14)C]LAC), [1-(14)C]pyruvic ([(14)C]PYR), or β-[1-(14)C]hydroxybutyric ([(14)C]BHB) was used to compare the oxidative metabolism of the end products with that of the KBs. Some cells were placed in a hypoxic chamber (1% O2). PAL and LC induced a higher production of KBs in astroglia than in neurons, while the CO2 production from PAL was less than 5% of the KB production in both astroglia and neurons. KB production in astroglia was augmented by the AMP-activated protein kinase activators, AICAR and metformin, as well as hypoxia with/without glucose. Neuronal KB production increased under hypoxia in the absence of PAL and LC. In neurons, [(14)C]LAC and [(14)C]PYR oxidation decreased after 24\u2009hr of hypoxia, while [(14)C]BHB oxidation was preserved. Astroglia responds to ischemia in\xa0vitro by enhancing KB production, and astroglia-produced KBs derived from fatty might serve as a neuronal energy substrate for the tricarboxylic cycle instead of lactate, as pyruvate dehydrogenase is susceptible to ischemia.© The Author(s) 2014 Reprints and permissions: sagepub.com/journalsPermissions.nav.

Keyword: glycolysis

Effects of proglycosyn (LY177507) on fatty metabolism in rat hepatocytes.

Proglycosyn (LY177507) belongs to a series of powerful agents that stabilize liver glycogen stores by promoting glycogen synthesis from different precursors and inhibiting glycogenolysis and . In the present study we have examined the effects of proglycosyn on fatty metabolism in isolated hepatocytes. Preincubation of hepatocytes with medium containing proglycosyn led to a marked inhibition of fatty synthesis de novo and acetyl-CoA carboxylase activity without affecting fatty synthase. Likewise, proglycosyn depressed the synthesis of triacylglycerols and phospholipids from labeled palmitate. Although octanoate oxidation was unaffected by proglycosyn, mitochondrial palmitate oxidation was notably stimulated. This effect may be attributed to the proglycosyn-induced decrease of intracellular malonyl-CoA levels relative to control incubations and the concomitant relieve of the inhibition of the mitochondrial-outer-membrane carnitine palmitoyl-transferase by malonyl-CoA. By contrast, neither peroxisomal palmitate oxidation nor peroxisomal carnitine palmitoyltransferase activity was changed upon hepatocyte incubation with proglycosyn. Results thus indicate that proglycosyn increases the fatty--oxidation efficiency of the liver at the expense of lipogenesis, and this may contribute to the proglycosyn-induced sparing of liver glycogen stores.

Keyword: glycolysis

Central role for aldose reductase pathway in myocardial ischemic injury.

Aldose reductase (AR), a member of the aldo-keto reductase family, has been implicated in the development of vascular and neurological complications of diabetes. Recently, we demonstrated that aldose reductase is a component of myocardial ischemic injury and that inhibitors of this enzyme protect rat hearts from ischemia-reperfusion injury. To rigorously test the effect of aldose reductase on myocardial ischemia-reperfusion injury, we used transgenic mice broadly overexpressing human aldose reductase (ARTg) driven by the major histocompatibility complex I promoter. Hearts from these ARTg or littermate mice (WT) (n=6 in each group) were isolated, perfused under normoxic conditions, then subjected to 50 min of severe low flow ischemia followed by 60 min of reperfusion. Creatine kinase (CK) release (a marker of ischemic injury) was measured during reperfusion; left ventricular developed pressure (LVDP), end diastolic pressure (EDP), and ATP were measured throughout the protocol. CK release was significantly greater in ARTg mice compared with the WT mice. LVDP recovery was significantly reduced in ARTg mice compared with the WT mice. Furthermore, ATP content was higher in WT mice compared with ARTg mice during ischemia and reperfusion. Infarct size measured by staining techniques and myocardial damage evaluated histologically were also significantly worse in ARTg mice hearts than in controls. Pharmacological inhibition of aldose reductase significantly reduced ischemic injury and improved functional recovery in ARTg mice. These data strongly support key roles for AR in ischemic injury and impairment of functional and metabolic recovery after ischemia. We propose that interventions targeting AR may provide a novel adjunctive approach to protect ischemic myocardium.

Keyword: glycolysis

Palmitoleic (n-7) increases white adipocytes GLUT4 content and glucose uptake in association with AMPK activation.

Palmitoleic was previously shown to improve glucose homeostasis by reducing hepatic glucose production and by enhancing insulin-stimulated glucose uptake in skeletal muscle. Herein we tested the hypothesis that palmitoleic positively modulates glucose uptake and metabolism in adipocytes.For this, both differentiated 3\xa0T3-L1 cells treated with either palmitoleic (16:1n7, 200\xa0μM) or (16:0, 200\xa0μM) for 24\xa0h and primary adipocytes from mice treated with 16:1n7 (300\xa0mg/kg/day) or oleic (18:1n9, 300\xa0mg/kg/day) by gavage for 10\xa0days were evaluated for glucose uptake, oxidation, conversion to lactate and incorporation into fatty acids and glycerol components of TAG along with the activity and expression of lipogenic enzymes.Treatment of adipocytes with palmitoleic, but not oleic (in vivo) or (in vitro) acids, increased basal and insulin-stimulated glucose uptake and GLUT4 mRNA levels and protein content. Along with uptake, palmitoleic enhanced glucose oxidation (aerobic ), conversion to lactate (anaerobic ) and incorporation into glycerol-TAG, but reduced de novo fatty synthesis from glucose and acetate and the activity of lipogenic enzymes glucose 6-phosphate dehydrogenase and ATP-citrate lyase. Importantly, palmitoleic induction of adipocyte glucose uptake and metabolism were associated with AMPK activation as evidenced by the increased protein content of phospho(p)Thr172AMPKα, but no changes in pSer473Akt and pThr308Akt. Importantly, such increase in GLUT4 content induced by 16:1n7, was prevented by pharmacological inhibition of AMPK with compound C.In conclusion, palmitoleic increases glucose uptake and the GLUT4 content in association with AMPK activation.

Keyword: glycolysis

THE INTERRELATIONSHIP BETWEEN GLUCOSE AND OXIDATION IN VITRO BY ASCITES HEPATOMA, AH 130.

Keyword: glycolysis

High levels of fatty acids increase contractile function of neonatal rabbit hearts during reperfusion following ischemia.

In the neonatal heart the transition from using carbohydrates to using fatty acids has not fully matured and oxidative metabolism/ATP generation may be limiting contractile function after ischemia. This study tested the hypothesis that increasing fatty availability increases recovery of left ventricular (LV) work by increasing palmitate oxidation, tricarboxylic (TCA) cycle activity, and ATP generation. Isolated working hearts from 7-day-old rabbits were perfused with Krebs solution containing low (0.4 mM) or high (2.4 mM) palmitate and 5.5 mM glucose. Hearts were subjected to 35-min global ischemia before 40-min reperfusion, and rates of , glucose oxidation, and palmitate oxidation were assessed. LV work was similar before ischemia but was greater during reperfusion in hearts perfused with 2.4 mM palmitate compared with hearts perfused with 0.4 mM palmitate [6.98 +/- 0.14 (n = 15) vs. 3.01 +/- 0.23 (n = 16) mJ.beat(-1).g dry wt(-1); P < 0.05]. This was accompanied by increased LV energy expenditure during reperfusion [35.98 +/- 0.16 (n = 8) vs. 19.92 +/- 0.18 (n = 6) mJ.beat(-1).g dry wt(-1); P < 0.05]. During reperfusion the rates of palmitate oxidation [237.5 +/- 28.10 (n = 7) vs. 86.0 +/- 9.7 (n = 6) nmol.g dry wt(-1).min(-1); P < 0.05], total TCA cycle activity [2.65 +/- 0.39 (n = 7) vs. 1.36 +/- 0.14 (n = 6) micromol acetyl-CoA.g dry wt(-1).min(-1); P < 0.05], and ATP generation attributable to palmitate oxidation [26.6 +/- 3.1 (n = 7) vs. 12.6 +/- 1.7 (n = 6) micromol.g dry wt(-1).min(-1); P < 0.05] were greater in hearts perfused with 2.4 mM palmitate. These data indicate that the neonatal heart has decreased energy reserve, and, in contrast to the mature heart, increasing availability of fatty substrate increases energy production and improves recovery of function after ischemia.

Keyword: glycolysis

Role of glycolytic flux in effect of glucose in decreasing fatty--induced release of lactate dehydrogenase from isolated coronary ligated rat heart.

The mechanisms whereby glucose reduces fatty -induced release of enzyme from the coronary-ligated isolated perfused working rat heart are investigated. Alterations in the tissue contents of ATP, phosphocreatine, or glycogen could be excluded as possible mechanisms for the beneficial effect of glucose in this system. Provision of glycolytic ATP from increased glycolytic flux may be one important factor.

Keyword: glycolysis

The contribution of , glucose oxidation, lactate oxidation, and fatty oxidation to ATP production in isolated biventricular working hearts from 2-week-old rabbits.

Immediately after birth, and lactate oxidation are the major source of ATP production in the rabbit heart. Although the ability of heart to oxidize fatty increases within days, glucose oxidation rates remain low in the first week after birth. To further examine the changes in energy substrate use in the newborn period, we developed a right and left ventricular isolated working heart model, in which 2-wk-old rabbit hearts were perfused with buffer containing 11 mM glucose, 0.8 mM palmitate, 0.5 mM lactate, and 100 microU/mL insulin. Hearts were perfused at a 7.5 mm Hg left atrial preload, a 4.5 mm Hg superior vena cava preload, a 30 mm Hg aortic afterload, and a 4.5 mm Hg pulmonary artery afterload. Glycolytic rates [measured as 3H2O production from (5-3H)-glucose] were 791 +/- 108 nmol/g dry weight.min-1 (mean +/- SEM). Oxidation rates of glucose, lactate, and palmitate (measured as 14CO2 production from 14C substrates) were 94 +/- 15, 126 +/- 13, and 60 +/- 8 nmol/g dry weight.min-1, respectively. In these hearts, the majority of ATP production derived from exogenous sources was obtained from fatty oxidation (52%), whereas 11, 23, and 15% of ATP requirements were derived from , glucose oxidation, and lactate oxidation, respectively. These studies demonstrate that by 2 wks of age in rabbits, fatty acids are the major source of energy in the heart. However, although the contribution of glucose oxidation to ATP production has increased compared with 1- or 7-d-old rabbit hearts, glucose oxidation rates are still low compared with adult hearts.

Keyword: glycolysis

NADPH oxidase-derived reactive oxygen species increases expression of monocyte chemotactic factor genes in cultured adipocytes.

Excess glucose and free fatty acids delivered to adipose tissue causes local inflammation, which contributes to insulin resistance. Glucose and palmitate generate reactive oxygen species (ROS) in adipocytes, leading to monocyte chemotactic factor gene expression. Docosahexaenoate (DHA) has the opposite effect. In this study, we evaluated the potential sources of ROS in the presence of excess nutrients. Differentiated 3T3-L1 adipocytes were exposed to palmitate and DHA (250 μM) in either 5 or 25 mM glucose to evaluate the relative roles of mitochondrial electron transport and NADPH oxidases (NOX) as sources of ROS. Excess glucose and palmitate did not increase mitochondrial oxidative phosphorylation. However, glucose exposure increased . Of the NOX family members, only NOX4 was expressed in adipocytes. Moreover, its activity was increased by excess glucose and palmitate and decreased by DHA. Silencing NOX4 inhibited palmitate- and glucose-stimulated ROS generation and monocyte chemotactic factor gene expression. NADPH, a substrate for NOX, and pentose phosphate pathway activity increased with glucose but not palmitate and decreased with DHA exposure. Inhibition of the pentose phosphate pathway by glucose-6-phosphate dehydrogenase inhibitors and siRNA suppressed ROS generation and monocyte chemotactic factor gene expression induced by both glucose and palmitate. Finally, both high glucose and palmitate induced NOX4 translocation into lipid rafts, effects that were blocked by DHA. Excess glucose and palmitate generate ROS via NOX4 rather than by mitochondrial oxidation in cultured adipocytes. NOX4 is regulated by both NADPH generated in the PPP and translocation of NOX4 into lipid rafts, leading to expression of monocyte chemotactic factors.

Keyword: glycolysis

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic metabolism-mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 diabetes.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: glycolysis

The role of fatty acids in ischemic tissue injury: difference between oleic and .

Guinea pig hearts were subjected to low-flow perfusion (0.3 ml/g fresh weight/min) with an oxygen depleted perfusate. Fatty acids ( or oleic ), added to the perfusate, accelerated in a dose-dependent manner the anoxic decay of creatine phosphate and ATP, impaired lactate production and augmented enzyme release (lactate dehydrogenase, malate dehydrogenase). and oleic , however, differed distinctly in their deleterious effect, this being greater for oleic . After 60 min anoxic low-flow perfusion with 11 mM glucose and 0.2 mM of either fatty , complexed in 5:1 molar relationship to albumin, the creatine phosphate content with palmitate is 39% greater than with oleate, the ATP content 23%, lactate production 15% greater, and release of malate dehydrogenase 24% lower, but the elevated contents of long-chain acyl CoA and acyl carnitine are not significantly different for the two fatty acids. These results accord with earlier experiences on subcellular systems showing that the physicochemical effects of the oleyl residue are more harmful than those of the palmityl residue.

Keyword: glycolysis

Exogenous substrate utilization by isolated myocytes from chronically diabetic rats.

The effect of chronic streptozotocin-induced diabetes on the utilization of exogenous substrates by freshly isolated, Ca2+-tolerant nonbeating myocytes was investigated. The rates of glucose (5 or 25 mM) and lactate (1 mM) oxidation were significantly reduced in myocytes of diabetic rats, whereas palmitate (0.4 or 1 mM) oxidation was similar to the controls. Glucose oxidation in diabetic (but not in control) and palmitate oxidation in control (but not in diabetic) myocytes were increased by raising the respective substrate concentrations in the medium to levels found in vivo in diabetic rats. Inhibition of glucose and lactate oxidation in the presence of competing substrates were generally similar between control and diabetic myocytes. However, the inhibitory effect of glucose on lactate oxidation was greater in control cells. The rate of palmitate oxidation was diminished by glucose in the controls, but this was not observed in the diabetic myocytes. Oxygen consumption by the myocytes of diabetic rats was below that of control cells when lactate or palmitate was present in the medium. ATP and phosphocreatine contents were similar in the myocytes of diabetic and control rats. All the observed changes in myocytes prepared from diabetic rats were reversed by in vivo insulin treatment.

Keyword: glycolysis

Lipid metabolism in pigs fed supplemental conjugated linoleic and/or dietary arginine.

We proposed that the combination of conjugated linoleic (CLA) and arginine would decrease adiposity by depressing lipid synthesis in liver and adipose tissues of growing pigs. Pigs were allotted to treatments in a 2 × 2 factorial design with two lipids (CLA or canola oil) and two amino acids [L-arginine or L-alanine (isonitrogenous control)]; supplements were provided from 80 to 110 kg body weight (approximately 4 weeks). Treatment groups (n = 4) were: control (2.05% L-alanine plus 1% canola oil); CLA (2.05% L-alanine plus 1% CLA); arginine (1.0% L-arginine plus 1.0% canola oil); arginine plus CLA (1.0% arginine plus 1.0% CLA). Arginine increased backfat thickness (P = 0.07) in the absence or presence of CLA, and arginine supplementation increased subcutaneous and retroperitoneal adipocyte volume, especially in combination with dietary CLA (interaction P = 0.001). Arginine increased palmitate incorporation into total lipids by over 60% in liver (P = 0.07). Dietary CLA increased palmitate incorporation into lipids in longissimus muscle by over 100% (P = 0.01), and CLA increased longissimus muscle lipid by nearly 20%. CLA increased glucose oxidation to CO(2) by over 80% in retroperitoneal and subcutaneous adipose tissues (P = 0.04), and doubled palmitate oxidation to CO(2) in intestinal duodenal mucosal cells (P = 0.07). Arginine supplementation decreased muscle pH at 45 min postmortem (P = 0.001), indicating elevated early postmortem , and CLA and arginine independently increased PGC-1α gene expression in longissimus muscle. CLA but not arginine depressed mTOR gene expression in intestinal duodenal mucosal cells. CLA decreased serum insulin by 50% (P = 0.02) but increased serum triacylglycerols by over 40%. CLA supplementation increased (P ≤ 0.01) total saturated fatty acids in liver and adipose tissue. In conclusion, neither CLA nor arginine depressed tissue lipid synthesis in growing/finishing pigs, and in fact dietary CLA promoted elevated intramuscular lipid and arginine increased carcass adiposity.

Keyword: glycolysis

Evaluation of human blood lymphocytes as a model to study the effects of drugs on human mitochondria. Effects of low concentrations of amiodarone on fatty oxidation, ATP levels and cell survival.

Human lymphocytes were assessed as a cellular model for determining the effects of drugs on human mitochondria. Formation of total oxidized 14C-products was maximal with 1 mM [U-14C], was linear for 90 min, linear with the number of lymphocytes, and decreased by 95% and 77% in the presence of 30 microM rotenone and 2 mM KCN. Seven drugs were tested which had previously been shown to inhibit beta-oxidation in animals; all decreased formation of total oxidized 14C-products by human lymphocytes, but with different IC50 values: 35 microM with amiodarone, 2.75 mM with tetracycline and amineptine, 3.75 mM with tianeptine, and more than 10 mM for valproic and the ibuprofen enantiomers. Formation of [14C]CO2 either increased or decreased, in relation to the various effects of these drugs on coupling, beta-oxidation, and the tricarboxylic cycle. There was a general trend for some relationship between inhibition of fatty oxidation and loss of cellular ATP. Those compounds, however, which uncoupled oxidative phosphorylation (2,4-dinitrophenol, amiodarone, ibuprofen) and/or inhibited the mitochondrial respiratory chain (amiodarone, rotenone, KCN) resulted in comparatively higher ATP depletion. Amiodarone, a drug which produces several effects (uncoupling, inhibition of beta-oxidation, of the tricarboxylic cycle and of the respiratory chain), caused a dramatic decrease in cellular ATP and cell viability at low concentrations (20-100 microM). Both these effects were prevented by the addition of 5 mM glucose, a substrate for anaerobic . We conclude that human lymphocytes may be a useful model for assessing the effects of drugs on human mitochondrial function. IC50 values determined with this model may not necessarily apply, however, to other cells.

Keyword: glycolysis

Capric Up-Regulates UCP3 Expression without PDK4 Induction in Mouse C2C12 Myotubes.

Uncoupling protein 3 (UCP3) and pyruvate dehydrogenase kinase 4 (PDK4) in skeletal muscle are key regulators of the glucose and lipid metabolic processes that are involved in insulin resistance. Medium-chain fatty acids (MCFAs) have anti-obesogenic effects in rodents and humans, while long-chain fatty acids (LCFAs) cause increases in body weight and insulin resistance. To clarify the beneficial effects of MCFAs, we examined UCP3 and PDK4 expression in skeletal muscles of mice fed a MCFA- or LCFA-enriched high-fat diet (HFD). Five-week feeding of the LCFA-enriched HFD caused high body weight gain and induced glucose intolerance in mice, compared with those in mice fed the MCFA-enriched HFD. However, the amounts of UCP3 and PDK4 transcripts in the skeletal muscle of mice fed the MCFA- or LCFA-enriched HFD were similar. To further elucidate the specific effects of MCFAs, such as capric (C10:0), on lipid metabolism in skeletal muscles, we examined the effects of various FAs on expression of UCP3 and PDK4, in mouse C2C12 myocytes. Although (C16:0) and lauric (C12:0) significantly induced expression of both UCP3 and PDK4, capric (C10:0) upregulated only UCP3 expression via activation of peroxisome proliferator-activated receptor-δ. Furthermore, (C16:0) disturbed the insulin-induced phosphorylation of Akt, while MCFAs, including lauric (C12:0), capric (C10:0), and caprylic (C12:0), did not. These results suggest that capric (C10:0) increases the capacity for fatty oxidation without inhibiting glycolysis in skeletal muscle.

Keyword: glycolysis

Effects of ethanol on brain metabolism.

The influence of acute or chronic ethanol administration on the biochemical processes in brain and cerebral metabolic pathways has been discussed. Ethanol seems to affect cerebral carbohydrate metabolism mainly through increased glycogenolysis, although the possibility of decreased cerebral glucose utilization remains eminent. Ethanol affects the consumption of oxygen by the brain tissue presumably through alterations in the brain cell membranes. Inhibition of Na+-K+-ATP-ase observed during ethanol intoxication is suspected to result in alterations in the membranes of the nerve cells. Isotope studies in addition to total respiratory carbon dioxide production strongly suggest the inhibition of citric cycle function during ethanol metabolism. Although synthetic pathways for lipids do not seem to be affected by ethanol, lipid oxidation in the cerebral tissue is significantly inhibited. In addition to above mentioned alterations in the cerebral metabolic processes, ethanol also affects ionic transport processes, adenine nucleotides, and amino and protein metabolism. The metabolic consequences of such effects of ethanol have been discussed.

Keyword: glycolysis

Short-term manipulation of plasma free fatty acids does not change skeletal muscle concentrations of ceramide and glucosylceramide in lean and overweight subjects.

Increased plasma free fatty (FFA) concentrations may be in part responsible for the increased levels of ceramide in skeletal muscle of obese subjects.We studied the effect of lowering and increasing plasma FFA levels on muscle ceramide and glucosylceramide concentrations in lean and obese subjects.Plasma FFAs were either increased or decreased for 6 h by infusing a lipid emulsion or using Acipimox, respectively. Muscle biopsies were performed before and after the intervention for measurements of ceramide and glucosylceramide.Eight lean [body mass index 21.9 (range, 19.6-24.6) kg/m2] and six overweight/obese [body mass index 34.4 (27.8-42.5) kg/m2] subjects without type 2 diabetes mellitus participated in the study.Differences in muscle ceramide and glucosylceramide upon manipulation of plasma FFAs were measured.There were no differences in muscle ceramide and glucosylceramide between lean and obese subjects, respectively. Increasing or decreasing plasma FFAs for 6 h had no effect on ceramide [high FFAs: 24 (19-25) vs. 24 (22-27) pmol/mg muscle, P=0.46; and 22 (20-28) vs. 24 (18-26) pmol/mg muscle, P=0.89 in lean and obese, respectively; low FFAs: 26 (24-35) vs. 23 (18-27) pmol/mg muscle, P=0.17 and 24 (15-44) vs. 24 (19-42) pmol/mg muscle, P=0.6 in lean and obese, respectively] and glucosylceramide [high FFAs: 2.0 (1.7-4.3) vs. 3.4 (2.1-4.6) pmol/mg muscle, P=0.17; and 3.0 (1.3-6.7) vs. 2.6 (1.2-3.9) pmol/mg muscle, P=0.89 in lean and obese, respectively; low FFAs: 2.2 (1.0-4.4) vs. 1.7 (1.4-3.0) pmol/mg muscle, P=0.92; and 6.6 (1.0-25.0) vs. 4.3 (1.3-7.6) pmol/mg muscle, P=0.7 in lean and obese, respectively] concentrations in skeletal muscle.Short-term manipulation of plasma FFAs has no effect on ceramide and glucosylceramide concentrations in skeletal muscle from lean and obese subjects.

Keyword: glycolysis

Computer simulation of metabolism in palmitate-perfused rat heart. II. Behavior of complete model.

Intermediary metabolism in rat hearts perfused with 11 mM glucose plus 1 mM palmitate was simulated by a computer model. Several enzyme submodels in a previous version of the isolated rat heart computer model were improved, and a new fatty oxidation pathway model was added. Compartmentation of metabolites in a pseudo-stationary state was calculated, and its implications are discussed, e.g., citrate level may not regulate because it is mostly mitochondrial. Citrate synthetase, controlled largely by its inhibitors, is of key importance in regulating fatty metabolism. The response of aconitase to the mitochondrial Mg2+ level is of major importance in setting both the mitochondrial citrate and isocitrate levels. Pyruvate dehydrogenase is about 96% in the inactive phosphorylated form, and the active form is also 15% inhibited by products, severely limiting pyruvate oxidation and causing preferential utilization of palmitate as the metabolic fuel. The simulation is consistent with a creatine phosphate shuttle which delivers high energy phosphate to the site of its utilization for mechanical work.

Keyword: glycolysis

Measurements of fatty and carbohydrate metabolism in the isolated working rat heart.

The isolated working rat heart is a useful experimental model which allows contractile function to be measured in hearts perfused at physiologically relevant workloads. To maintain these high workloads the heart is required to generate a tremendous amount of energy. In vivo this energy is derived primarily from the oxidation of fatty acids. In many experimental situations it is desirable to perfuse the isolated working heart in the presence of physiologically relevant concentrations of fatty acids. This is particularly important when studying energy metabolism in the heart, or in determining how fatty acids alter the outcome of myocardial ischemic injury [1, 2]. The other major source of energy for the heart is derived from the oxidation of carbohydrates (glucose and lactate), with a smaller amount of ATP also being derived from . Two byproducts of both fatty and carbohydrate metabolism are H2O and CO2. By labeling the glucose, lactate, or fatty acids in the perfusate with 3H or 14C the experimenter can quantitatively collect either 3H2O or 14CO2 produced by the heart. By using radioisotopes that are labeled at specific hydrogen or carbon molecules on the various energy substrates, and by knowing the specific activity of the radiolabeled substrate used, it is possible to determine the actual rate of flux through these individual pathways. This paper will describe the experimental protocols for directly measuring fatty and carbohydrate metabolism in isolated working rat hearts.

Keyword: glycolysis

[Planar scintigraphy versus PET in measuring fatty metabolism of the heart].

In addition to radionuclide ventriculography and thallium scintigraphy, already well established in nuclear medicine, assessment of myocardial metabolism is also of interest for diagnosis and follow-up observations in heart disease. Under aerobic conditions and in the fasting state, the heart muscle primarily oxidizes fatty acids; during ischemia, in contrast, there is slowing of fatty turnover and increased anaerobic . With 11C-, in humans, reduced fatty uptake has been documented in infarcted myocardial regions. The analysis of 11C- in dogs showed a three-phased elimination curve in normal myocardium. In ischemic myocardium, there was diminished utilization of free fatty acids and the glucose utilization was concomitantly increased. After insulin-glucose infusion, as well, there was increased glucose utilization and a reduction in fatty utilization. Studies with 11C- require the equipment for positron emission tomography (PET); because of the short half-life of 20.3 minutes, the nuclide must be generated by a cyclotron in the immediate vicinity. In the search for well-suited isotopes for use in planar scintigraphy employing a gamma camera, the uptake and elimination of a variety of isotopically-marked fatty acids were measured and compared with the characteristics of 14C-. For 17-123I-heptadecanic (IHA) the elimination curve was similar to that of 14C-palmitate: disadvantage, however, was the relatively high percentage of water soluble marked catabolites which required dual parameter analysis by means of 99-m-technetium pertechnetate or 123I sodium iodide to quantify the amount of myocardial fatty utilization through subtraction of the externally measured water soluble catabolite from the externally measured total activity. In studies with the gamma camera in fasting patients in whom 2 to 3 mCi IHA was injected intravenously after symptom limited bicycle ergometry, in healthy subjects the elimination halftime for the first rapid phase was 24.4 +/- 4.7 minutes. Patients with angiographically-documented coronary artery disease, in the afflicted myocardial segments, had diminished fatty uptake and prolonged elimination halftime as compared with normally perfused segments. In patients with dilated cardiomyopathy there was an inhomogeneous distribution of activity in the myocardium and, in contrast to coronary artery disease, a discordance between local fatty uptake and turnover rate. After chronic and acute alcohol consumption there were comparable findings which were shown to be reversible after several weeks of abstinence.(ABSTRACT TRUNCATED AT 400 WORDS).

Keyword: glycolysis

Sodium stibogluconate (Pentostam) inhibition of glucose catabolism via the glycolytic pathway, and fatty beta-oxidation in Leishmania mexicana amastigotes.

The biochemical mechanism of action of antimony (Sb) in pentavalent form complexed to gluconic (sodium stibogluconate)--the drug of choice for the leishmaniases--has been only slightly investigated. We recently reported that, in stibogluconate-exposed Leishmania mexicana amastigotes, there is a dose-dependent decrease in the ATP/ADP ratio [Berman et al., Antimicrob. Agents Chemother. 27, 916 (1985)]. To investigate mechanisms by which ADP phosphorylation to ATP might be inhibited, stibogluconate-exposed amastigotes were incubated with [14C]glucose, fatty , or acetate, and 14CO2 production was determined. In organisms exposed to 500 micrograms Sb/ml, formation of 14CO2 from [6-14C]glucose and [1-14C]palmitate was inhibited 69 and 67% respectively. In comparison, formation of 14CO2 from [1-14C]glucose and [2-14C]acetate was inhibited less than 15%. These results suggest that glucose catabolism via glycolytic enzymes and fatty beta-oxidation, but not glucose metabolism via the hexosemonophosphate shunt or the citric cycle, is specifically inhibited in stibogluconate-exposed Leishmania mexicana amastigotes. Inhibition of these pathways suggests a mechanism for the inhibition of ADP phosphorylation previously reported.

Keyword: glycolysis

Purinergic inhibition of glucose transport in cardiomyocytes.

ATP is known to act as an extracellular signal in many organs. In the heart, extracellular ATP modulates ionic processes and contractile function. This study describes a novel, metabolic effect of exogenous ATP in isolated rat cardiomyocytes. In these quiescent (i.e. noncontracting) cells, micromolar concentrations of ATP depressed the rate of basal, catecholamine-stimulated, or insulin-stimulated glucose transport by up to 60% (IC50 for inhibition of insulin-dependent glucose transport, 4 microM). ATP decreased the amount of glucose transporters (GLUT1 and GLUT4) in the plasma membrane, with a concomitant increase in intracellular microsomal membranes. A similar glucose transport inhibition was produced by P2 purinergic agonists with the following rank of potencies: ATP approximately ATPgammaS approximately 2-methylthio-ATP (P2Y-selective) > ADP > alpha,betameATP (P2X-selective), whereas the P1 purinoceptor agonist adenosine was ineffective. The effect of ATP was suppressed by the poorly subtype-selective P2 antagonist pyridoxal-phosphate-6-azophenyl-2\', 4\'-disulfonic but, surprisingly, not by the nonselective antagonist suramin nor by the P2Y-specific Reactive Blue 2. Glucose transport inhibition by ATP was not affected by a drastic reduction of the extracellular concentrations of calcium (down to 10(-9) M) or sodium (down to 0 mM), and it was not mimicked by a potassium-induced depolarization, indicating that purinoceptors of the P2X family (which are nonselective cation channels whose activation leads to a depolarizing sodium and calcium influx) are not involved. Inhibition was specific for the transmembrane transport of glucose because ATP did not inhibit (i) the rate of under conditions where the transport step is no longer rate-limiting nor (ii) the rate of [1-14C]pyruvate decarboxylation. In conclusion, extracellular ATP markedly inhibits glucose transport in rat cardiomyocytes by promoting a redistribution of glucose transporters from the cell surface to an intracellular compartment. This effect of ATP is mediated by P2 purinoceptors, possibly by a yet unknown subtype of the P2Y purinoceptor family.

Keyword: glycolysis

Dominant uptake of fatty over glucose by prostate cells: a potential new diagnostic and therapeutic approach.

Prostate cancer is characterized by a low rate of and glucose uptake. We hypothesize that fatty is dominant over glucose in uptake by prostate cells.One benign (RWPE1) and two malignant (LNCaP and PC3) prostate cell lines were assayed for their in vitro uptake of radiolabeled glucose analogs (3)H-fluoro-2-deoxyglucose and (18)F-fluoro-2-deoxyglucose, and the long-chain fatty (3)H-. Fractional uptake was standardized to viable cell numbers.Uptake of palmitate in all 3 prostate cell lines was significantly higher than that of glucose at all incubation times (p<0.01). But in malignant cell lines, neither glucose nor palmitate uptake was quantitatively higher than that in the benign cell line. The uptake of fatty by prostate cells is a dynamic, active process mediated by the membrane receptors.Prostate cells are characterized by a dominant uptake of fatty over glucose, suggesting that future development of new diagnostic and therapeutic approaches in prostatic cancer should focus on fatty substrate. Fatty imaging may be useful in detection of recurrence and metastasis, but not in differentiating malignant from benign prostate tissue.

Keyword: glycolysis

Metabolism of palmitate by anoxic and reoxygenated heart cell cultures.

Under anoxic incubation conditions heart cell cultures showed enhanced uptake of [U-14C] into neutral lipids, while incorporation into phospholipids was unaltered. Fractionation of the neutral lipids showed greatest incorporation of radiolabel into the triglyceride fraction. Uptake of fatty in normoxic cultures may be dependent upon the supply of glycerol 3-phosphate from , as 2-deoxyglucose and L-lactate, respectively, inhibited and stimulated incorporation of fatty into neutral lipid fractions. When previously anoxic cultures were reoxygenated, oxidation of fatty was depressed and the mitochondrial function of anoxic cultures appeared to be more readily uncoupled by 2,4-dinitrophenol, in comparison with cultures maintained under normoxic conditions. Similar behaviour was seen when oxidation of endogenous lipid or oxidation of glucose was examined. Previously anoxic cultures show a preference for oxidation of endogenous rather than exogenous lipid substrates. The results suggest that anoxia-stimulated lipid accumulation may prove injurious to subsequent mitochondrial function and may be a contributory factor in the pathological processes associated with hypoxic injury of cardiac tissue.

Keyword: glycolysis

Glucose and lipid metabolism disorders in the chickens with dexamethasone-induced oxidative stress.

The purpose of this study was to investigate the effects of long-term treatment with dexamethasone (DEX) on the antioxidation and nutrition metabolism in broiler chickens. Broilers were placed on a high-nutrient diet for 41\xa0days, and half were given orally DEX-supplemented water at 20\xa0mg/L every other day from 19 to 41\xa0days of age. DEX treatment downregulated superoxide dismutase activity as well as the mRNA expression of CuZn-superoxide dismutase and glutathione peroxidase with a decrease in GSH/GSSG ratio and an increase in malondialdehyde level in the liver of broilers. DEX treatment aggravated oxidative damage in the liver and, therefore, increased the sensitivity of broilers to ascites syndrome with higher mortality and reduced growth performance. Serum metabolomics analysis showed that DEX treatment significantly increased the levels of glucose, intermediates in protein metabolism (valine, proline, serine, threonine and urea) and lipid metabolism-related products (, stearic and cholesterol) while decreasing the levels of β-hydroxy butyric , succinic and malic , demonstrating that DEX treatment inhibited the Krebs cycle and the oxidation of fatty acids, and promoted the de novo synthesis of fatty acids as well as protein decomposition in the liver of broilers. Additionally, detection of metabolism-related enzymes\xa0revealed\xa0that DEX treatment inhibited and promoted glycogen decomposition. In summary, DEX treatment resulted in oxidative stress and glucose and lipid\xa0metabolism disorders in the broilers.© 2017 Blackwell Verlag GmbH.

Keyword: glycolysis

Fatty -induced nuclear translocation of heparanase uncouples glucose metabolism in endothelial cells.

Heparanase is an endoglycosidase that specifically cleaves carbohydrate chains of heparan sulfate. We have recently reported that high fatty increased the nuclear content of endothelial heparanase. Here, we examined the mechanism and the consequences behind this nuclear translocation of heparanase.Bovine coronary artery endothelial cells were grown to confluence and incubated with . induced rapid nuclear accumulation of heparanase that was dependent on Bax activation and lysosome permeabilization. Heat shock protein 90 was an important mediator of -induced shuttling of heparanase to the nucleus. Nuclear heparanase promoted cleavage of heparan sulfate, a potent inhibitor of histone acetyltransferase activity and gene transcription. A TaqMan gene expression assay revealed an increase in genes related to glucose metabolism and inflammation. In addition, was uncoupled from glucose oxidation, resulting in accumulation of lactate.The results presented in this study demonstrate that fatty can provoke lysosomal release of heparanase, its nuclear translocation, activation of genes controlling glucose metabolism, and accumulation of lactate. Given that lactate and inflammation have been implicated in the progression of atherosclerosis, our data may serve to reduce the associated cardiovascular complications seen during diabetes.

Keyword: glycolysis

Palmitate-induced changes in protein expression of insulin secreting INS-1E cells.

Elevated blood levels of glucose and lipids in individuals with type 2 diabetes mellitus have been observed to cause impairment of insulin secretion from pancreatic beta-cells. Chronic exposure to either of the circulating fatty oleate or palmitate has different effects on the beta-cell. Whereas palmitate causes functional impairment of the beta-cell and apoptosis, oleate has only minor negative effects on beta-cell function and mass. The aim of the present study was to delineate mechanisms by which the fatty acids affect the beta-cell differently. In particular, the aim was to identify beta-cell proteins exclusively regulated by palmitate. INS-1E cells were cultured for 24h in medium supplemented with palmitate or oleate. After culture, cells were lysed and subjected to two-dimensional gel electrophoresis. Proteins specifically regulated by palmitate were excised from the gel and identified by peptide mass fingerprinting using MALDI-TOF MS. Proteins exclusively regulated by palmitate were classified into proteins of carbohydrate or protein metabolism and Ca(2+) or mRNA binding proteins. The specific palmitate-induced down-regulation of enzymes of , proteins of protein turnover and anti-apoptotic protein may contribute to explain the different effects exerted by palmitate and oleate on beta-cell function and mass.Copyright 2010 Elsevier B.V. All rights reserved.

Keyword: glycolysis

Regulation of mammalian pyruvate dehydrogenase.

In mammalian tissues, two types of regulation of the pyruvate dehydrogenase complex have been described: end product inhibition by acetyl CoA and NADH: and the interconversion of an inactive phosphorylated form and an active nonphosphorylated form by an ATP requiring kinase and a specific phosphatase. This article is largely concerned with the latter type of regulation of the complex in adipose tissue by insulin (and other hormones) and in heart muscle by lipid fuels. Effectors of the two interconverting enzymes include pyruvate and ADP which inhibit the kinase, acetoin which activates the kinase and Ca2+ and Mg2+ which both activate the phosphatase and inhibit the kinase. Evidence is presented that all components of the pyruvate dehydrogenase complex including the phosphatase and kinase are located within the inner mitochondrial membrane. Direct measurements of the matrix concentration of substrates and effectors is not possible by techniques presently available. This is the key problem in the identification of the mechansims involved in the alterations in pyruvate dehydrogenase activity observed in adipose tissue and muscle. A number of indirect approaches have been used and these are reviewed. Most hopeful is the recent finding in this laboratory that in both adipose tissue and heart muscle, differences in activity of pyruvate dehydrogenase in the intact tissue persist during preparation and subsequent incubation of mitochondria.

Keyword: glycolysis

Comparison of the oxidation of glutamine, glucose, ketone bodies and fatty acids by human diploid fibroblasts.

The contribution of glutamine, glucose, ketone bodies and fatty acids to the oxidative energy metabolism of human diploid fibroblasts ws studied. The rate of glutamine oxidation by fibroblasts was 98 nmol/h per mg cell protein compared to 2 nmol/h per mg cell protein or less for glucose, acetoacetate, D-3-hydroxybutyrate, octanoic and . Glucose inhibited glutamine oxidation by 85%, while the other substrates had no effect. Therefore, these cells meet their energy requirement almost solely by anaerobic and glutamine oxidation.

Keyword: glycolysis

Enhanced myocardial preservation by nicotinic , an antilipolytic compound: mechanism of action.

The cardioprotective effects of an antilipolytic compound, nicotinic , on arrested-reperfused myocardium were investigated in the isolated in situ pig heart preparation. Hearts were preperfused for 15 min in the presence of (5-3H)-glucose and (U-14C)-. Half of the hearts were then perfused with 0.08 mM nicotinic for an additional 15-min period, while the remaining control hearts received unmodified perfusion. Arrest was then induced in all animals for 2 h using hypothermic K+ cardioplegia, followed by 60 min of normothermic reperfusion. In control hearts, there were significantly greater levels of long-chain acyl Co-A and acyl carnitine and lower levels of membrane phospholipids than in the nicotinic group. While nicotinic inhibited beta-oxidation during pre-ischemia and reperfusion, it also prevented the degradation of membrane phospholipids. The net result was a reduction of free fatty accumulation during arrest and reperfusion in the nicotinic group. , as reflected in 3H2O production, was significantly increased by nicotinic administration. In the control heart as compared to the nicotinic group, the incorporation of 14C-label from palmitate into triglyceride and cholesterol during arrest was enhanced, while incorporation into phospholipids was depressed. The cardioprotective effects of nicotinic were demonstrated by decreased release of creatine kinase and improved coronary blood flow, and cardiac contractility in the reperfused myocardium supplemented with nicotinic compared to the control group. These results suggest that nicotinic significantly protects the arrested-reperfused myocardium by a) preventing elevation of myocardial fatty levels, b) stimulating by limiting fatty oxidation, c) inhibiting degradation of membrane phospholipids, and d) preventing accumulation of fatty metabolites with harmful detergent properties.

Keyword: glycolysis

The influence of a novel propolis on mutans streptococci biofilms and caries development in rats.

A flavonoids-free Brazilian propolis (type 6) showed biological effects against mutans streptococci and inhibited the activity of glucosyltransferases. This study evaluated the influence of the ethanolic extract of a novel type of propolis (EEP) and its purified hexane fraction (EEH) on mutans streptococci biofilms and the development of dental caries in rats. The chemical composition of the propolis extracts were examined by gas chromatography/mass spectrometry. The effects of EEP and EEH on Streptococcus mutans UA159 and Streptococcus sobrinus 6715 biofilms were analysed by time-kill and glycolytic pH drop assays. Their influence on proton-translocating F-ATPase activity was also tested. In the animal study, the rats were infected with S. sobrinus 6715 and fed with cariogenic diet 2000. The rats were treated topically twice a day with each of the extracts (or control) for 5 weeks. After the experimental period, the microbial composition of their dental plaque and their caries scores were determined. The results showed that fatty acids (oleic, , linoleic and stearic) were the main compounds identified in EEP and EEH. These extracts did not show major effects on the viability of mutans streptococci biofilms. However, EEP and EEH significantly reduced production by the biofilms and also inhibited the activity of F-ATPase (60-65%). Furthermore, both extracts significantly reduced the incidence of smooth surface caries in vivo without displaying a reduction of the percentage of S. sobriuns in the animals\' plaque (P < 0.05). However, only EEH was able to reduce the incidence and severity of sulcal surface caries (P < 0.05). The data suggest that the cariostatic properties of propolis type 6 are related to its effect on production and tolerance of cariogenic streptococci; the biological activities may be attributed to its high content of fatty acids.

Keyword: glycolysis

Effects of Stanniocalcin-1 on glucose flux in rat brown adipose tissue.

The present work assesses in\xa0vitro the role of human Stanniocalcin 1 (hSTC-1) in C-glucose metabolism in brown adipose tissue (BAT) from fed rat. In the fed state, hSTC-1 decreases the incorporation of C from glucose into lipids in the rat BAT. The data support the hypothesis that the capacity of the glycerol-3-phosphate (G3P)-generating pathway (glycolysis) from glucose is regulated by hSTC-1, decreasing the adequate supply of G3P needed for fatty esterification and triacylglycerol (TG) storage in BAT. The results also suggest the effect of hSTC-1 on de novo fatty synthesis from pyruvate generated by C-glucose in the glycolysis pathway. In addition, by decreasing lipogenesis, hSTC-1 increased ATP levels and these two factors may decrease BAT thermogenic function. The presence of hSTC-1 in the incubation medium did not alter C-glucose and C-1- oxidation. The uncoupling protein 1 (UCP-1) expression was not altered by hSTC-1 either. In conclusion, hSTC-1 is one of the hormonal factors that control glucose metabolism in BAT in the fed state. The decrease of TG capacity synthesis from C-glucose by hSTC-1 compromises the BAT thermogenic capacity. Furthermore, the increase in ATP levels would inhibit a futile cycle via UCP-1, which dissipates oxidative as heat.Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: glycolysis

Control of oxidative metabolism in volume-overloaded rat hearts: effect of propionyl-L-carnitine.

The objective of the present work was the assessment of metabolic events responsible for the improvement of hemodynamic function of volume-overloaded hearts from rats receiving propionyl-L-carnitine. A severe cardiac hypertrophy was induced in 2-mo-old rats by surgical opening of an aortocaval communication. Three months later, during in vitro perfusions with 1.2 mM palmitate, 11 mM glucose, and 10 IU/l insulin, the mechanical performance and overall energy turnover (myocardial O2 consumption) of hypertrophied rat hearts were significantly decreased under conditions of moderate and high workloads. These changes in cardiac energetics paralleled the decrease in total tissue carnitine content and alterations in exogenous palmitate oxidation. The oxidative utilization of glucose was also slightly depressed in volume-overloaded hearts while steady-state rates increased, especially in hearts subjected to high mechanical loads. This slowing of metabolic pathways involved in acetyl-CoA generation resulted in decreased NADH availability and in an apparent substrate limitation of oxidative phosphorylation suggested by a failure of cytosolic unbound ADP to drive respiration. Long-term administration of propionyl-L-carnitine normalized the degree of reduction of mitochondrial pyridine nucleotides and improved the kinetics of mitochondrial ATP production in volume-overloaded hearts. The resulting acceleration of energy turnover was essentially related to improved oxidative utilization of glucose, but steady-state palmitate oxidation rates also increased in severely hypertrophied hearts. This concomitant acceleration of glucose and palmitate oxidation may be related to the particular experimental conditions (high exogenous palmitate concentrations, elevated workloads) used in this study. We assume that the increase in intracellular carnitine, together with a stimulation of acetyl-CoA demands related to high workloads, creates conditions that are compatible with the simultaneous relief of pyruvate dehydrogenase and carnitine palmitoyltransferase I. The resulting increase in the rate of steady-state ATP production improves, in turn, the mechanical activity of volume-overloaded hearts.

Keyword: glycolysis

Liver composition and lipid metabolism in NZB/W F1 female mice fed dehydroisoandrosterone.

The beneficial effects obtained with dehydroisoandrosterone (DHA) feeding in the treatment of murine systemic lupus erythematosus are similar to those obtained with caloric restriction or with dietary manipulation of essential fatty availability. In this study, the fatty composition of selected tissues was examined in NZB/W F1 mice fed a diet containing 0.4% DHA. The effect of the DHA diet on liver composition and the activity of key hepatic enzymes involved in fatty synthesis and glucose metabolism was also investigated. The content of the essential fatty , arachidonate, was decreased in plasma cholesteryl esters and liver and kidney phospholipids in mice fed the DHA diet, yet no significant decrease in arachidonate content was observed in plasma phospholipid. The most striking change in both plasma and liver phospholipid was an increase in and a decrease in stearic , which could result from a decreased ability for fatty elongation. The liver mass was dramatically increased in the mice fed DHA, primarily from parenchymal cell hypertrophy, and contained little lipid. Significant changes in the activities of malic enzyme, glucose-6-phosphate dehydrogenase and pyruvate kinase, similar to those changes which occur with fasting, were observed during the initial adaptation to the DHA diet. The pyruvate kinase activity remained low, suggesting a decrease in liver . These results are consistent with the concept that diets containing DHA result in an altered metabolism with a decreased dependence on carbohydrate metabolism and an increased metabolism of lipids.

Keyword: glycolysis

Acute regulation of fatty oxidation and amp-activated protein kinase in human umbilical vein endothelial cells.

It is generally accepted that endothelial cells generate most of their ATP by anaerobic and that very little ATP is derived from the oxidation of fatty acids or glucose. Previously, we have reported that, in cultured human umbilical vein endothelial cells (HUVECs), activation of AMP-activated protein kinase (AMPK) by the cell-permeable activator 5-aminoimidazole-4-carboximide riboside (AICAR) is associated with an increase in the oxidation of (3)H-palmitate. In the present study, experiments carried out with cultured HUVECs revealed the following: (1) AICAR-induced increases in palmitate oxidation during a 2-hour incubation are associated with a decrease in the concentration of malonyl coenzyme A (CoA) (an inhibitor of carnitine palmitoyl transferase 1), which temporally parallels the increase in AMPK activity and a decrease in the activity of acetyl CoA carboxylase (ACC). (2) AICAR does not stimulate either palmitate oxidation when carnitine is omitted from the medium or oxidation of the medium-chain fatty octanoate. (3) When intracellular lipid pools are prelabeled with (3)H-palmitate, the measured rate of palmitate oxidation is 3-fold higher, and in the presence of AICAR, it accounts for nearly 40% of calculated ATP generation. (4) Incubation of HUVECs in a glucose-free medium for 2 hours causes the same changes in AMPK, ACC, malonyl CoA, and palmitate oxidation as does AICAR. (5) Under all conditions studied, the contribution of glucose oxidation to ATP production is minimal. The results indicate that the AMPK-ACC-malonyl CoA-carnitine palmitoyl transferase 1 mechanism plays a key role in the physiological regulation of fatty oxidation in HUVECs. They also indicate that HUVECs oxidize fatty acids from both intracellular and extracellular sources, and that when this is taken into account, fatty acids can be a major substrate for ATP generation. Finally, they suggest that AMPK is likely to be a major factor in modulating the response of the endothelium to stresses that alter its energy state.

Keyword: glycolysis

Effect of palmitate on carbohydrate utilization and Na/K-ATPase activity in aortic vascular smooth muscle from diabetic rats.

Several investigators have reported that carbohydrate metabolism is suppressed in blood vessels from diabetic (Db) rats. However, it is not known if metabolites from the reciprocal increase in oxidation of long-chain fatty acids that accompanies insulin-deficiency exacerbates the suppression of this pathway in the Db blood vessels. Such inhibition may have particularly deleterious consequences in vascular smooth muscle since aerobic is believed to preferentially fuel the sarcolemmal Na/K ATPase in this tissue. Therefore, this study evaluated the effect of physiological (0.4 mM) and elevated (1.2 mM) concentrations of the long-chain fatty palmitate on both carbohydrate utilization and Na/K-ATPase activity in aorta from insulin-deficient Db rat. Thoracic aorta were removed from 10 week Db (streptozotocin 60 mg/Kg , i.v.) or control (C) rats and intima-media aortic preparations were incubated in the absence or presence of palmitate. (microM/g dry wt/h) and glucose oxidation (microM/g dry wt/h) were quantified using 3H-glucose and 14C-glucose, respectively. Na/K-ATPase activity was estimated by the measurement of 86rubidium uptake in the absence and presence of 2 mM ouabain. In the absence of exogenous palmitate, (p < 0.05), glucose oxidation (p < 0.01) and the estimated ATP production from exogenous glucose were decreased in aorta from Db rat. However, despite this diminished rate of , Na/K ATPase activity was similar in Db and C aorta. Palmitate (0.4 mM) inhibited Na/K ATPase activity and glucose oxidation to a similar extent in both Db and C but had no effect on in either group. Elevation of palmitate to 1.2 mM had no additional inhibitory effect on glucose oxidation, Na/K ATPase activity or in either the Db or C aorta. The metabolism of exogenous palmitate restored the ATP production in Db to control values. These data demonstrate that, despite the diminished and glucose oxidation demonstrated in the Db tissue, Na/K ATPase activity was comparable in the C and Db aorta, in the absence or presence of exogenous long-chain fatty . Therefore, the accelerated oxidation of palmitate in diabetic vascular smooth muscle had no additional inhibitory effect on or Na/K ATPase activity. These data suggest that Na/K ATPase activity in vascular smooth muscle is not impaired by the altered pattern of substrate utilization that occurs in insulin-deficient Db rats.

Keyword: glycolysis

Palmitate-induced cardiac apoptosis is mediated through CPT-1 but not influenced by glucose and insulin.

To test the hypothesis that regulation of palmitate metabolism, through carnitine palmitoyl transferase-1 (CPT-1) or through alterations of , was involved in the pathway of palmitate-mediated cell death, cardiomyocytes were cultured from 7-day-old chick embryos. Palmitate-induced cell death, assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, was enhanced by carnitine, a cofactor needed for palmitate transport into mitochondria via CPT-1. Carnitine co-incubation with palmitate significantly (P < 0.01) increased the amount of apoptotic cells, assessed by propidium iodine staining and fluorescent-activated cell sorting analysis compared with treatment with either palmitate or carnitine alone. The CPT-1 inhibitor oxfenicine significantly (P < 0.05) blocked the cell death induced by the combination of palmitate and carnitine. The short-chain saturated fatty capric (100 microM), which is not likely transported by CPT-1, did not significantly affect cell viability, whereas the C18 saturated fatty stearic (100 microM) significantly (P < 0.01) reduced cell viability and to a similar extent as palmitate. In contrast, there was no significant alteration of palmitate-induced cell death by cotreatment with 100 nM insulin + 2 g/l glucose or 1 mM lactate, which promote ATP generation by rather than fatty oxidation. Fumonisin did not alter palmitate-induced cell death or apoptosis, suggesting that the effect of palmitate was not operative through increased ceramide synthesis. These results suggest that oxidation of palmitate through CPT-1 is involved in the production of apoptosis in cardiomyocytes.

Keyword: glycolysis

Mechanism of antitumor action of dimethanesulfonylthiopentane.

Keyword: glycolysis

miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and . In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: glycolysis

Isolation of intestinal mucosa cells.

Keyword: glycolysis

Effects of endoglycoceramidase or D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol on glucose uptake, , and mitochondrial respiration in HL60 cells.

The glycosphingolipid content of HL60 cells was reduced by endoglycoceramidase, an enzyme which specifically hydrolyzes glycosphingolipids on the cell surface, or by D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol, an inhibitor which specifically reduces the activity of UDP-glucose:ceramide glucosyltransferase. Reduction of the glycosphingolipid content by both reagents resulted in enhancement of glucose uptake and . Neither of these effects was observed in the presence of cytochalasin B, an inhibitor of facilitated glucose transport. The uptake of radiolabeled 3-O-methylglucose by the cells was not affected by treatment with either of the reagents, indicating no activation of the glucose transporter. On the other hand, both reagents decreased the level of ATP and CO2 production. The molecule mediating these effects appeared to be ceramide, since both treatments actually increased the intracellular ceramide content, and the cell-permeable short-chain ceramide N-acetylsphingosine, but not sphingosine, sphinganine, or , mimicked the effects of both reagents to comparable extents. Finally, the function of electron transport in isolated mitochondria fractions was found to be reduced by treatment of the cells with N-acetylsphingosine. These results strongly suggest that ceramide may affect mitochondrial respiration.Copyright 1998 Academic Press.

Keyword: glycolysis

Acetyl-CoA carboxylase regulation of fatty oxidation in the heart.

The role of acetyl-coenzyme A carboxylase (ACC) in regulating fatty oxidation was investigated in isolated fatty perfused working rat hearts. Overall fatty oxidation rates were determined by addition of 1.2 mM [3H]palmitate to the perfusate of hearts in which the endogenous triglyceride pool was prelabeled with [14C]palmitate. Rates of both exogenous and endogenous fatty oxidation were measured by simultaneous measurement of 3H2O and 14CO2 production, respectively. A second series of hearts were perfused under similar conditions except that [U-14C]glucose was present in the perfusate for measurement of glucose oxidation rates. Addition of dichloroacetate (DCA, 1 mM) to the perfusate resulted in a dramatic stimulation of glucose oxidation (a 411% increase), with a parallel decrease in fatty oxidation (from 305 +/- 51 to 206 +/- 40 nmol/g dry weight.min.unit work). DCA treatment increased the contribution of glucose oxidation to ATP production from 7.1 to 30.6%, while decreasing the contribution of overall fatty oxidation from 92.9 to 69.4%. Tissue levels of malonyl-CoA in hearts treated with DCA were higher compared to controls (14.0 +/- 0.6 and 10.0 +/- 0.7 nmol/g dry weight, respectively) and were negatively correlated (r = -0.85) with overall fatty oxidation rates. Acetyl-CoA levels were also significantly higher in DCA-treated hearts, and a positive correlation (r = 0.88) was seen between myocardial acetyl-CoA and malonyl-CoA levels. This suggests that DCA treatment increased the supply of acetyl-CoA for ACC. Western blots revealed the presence of both the 280-kDa (ACC-280) and the 265-kDa (ACC-265) isoforms of ACC in cardiac tissue, with a predominance of ACC-280. The activity of ACC extracted from hearts was similar in both groups when assayed under optimal conditions of acetyl-CoA and citrate. However, using affinity purified ACC, it was demonstrated that heart ACC (predominantly ACC-280) had a higher Km for acetyl-CoA than ACC isolated from white adipose tissue (predominantly ACC-265). We conclude that ACC is an important regulator of fatty oxidation in the heart and that acetyl-CoA supply is a key determinant of heart ACC-280 activity. As acetyl-CoA levels increase, ACC-280 is activated resulting in an increase in malonyl-CoA inhibition of fatty oxidation.

Keyword: glycolysis

Computer simulation of metabolism in palmitate-perfused rat heart. III. Sensitivity analysis.

The behavior of a computer model of metabolism in glucose- and palmitate-perfused rat hearts was interpreted by sensitivity analysis to explain why the heart preferentially utilizes fatty acids as fuel even in the presence of substantial exogenous glucose. The sensitivity functions identified those metabolites and enzymes which were most important in regulating the metabolic rate and determined which enzymes set the levels of the critical metabolites. Control of the mitochondrial redox potential and the distribution of coenzyme A thioesters regulated the rate of fatty utilization while strong inhibition of citrate synthetase resulted in accumulation of acetyl CoA and suppression of pyruvate oxidation. was limited by the cytosolic ATP/ADP ratio set largely by the creatine shuttle. Metabolic control appears to be widely distributed rather than localized at "key" enzymes. Metabolite levels are usually set by enzymes controlled by modifiers whereas metabolic flux is regulated by the enzymes that produce ligands for the modifier-controlled enzymes.

Keyword: glycolysis

Chronic activation of PPARalpha is detrimental to cardiac recovery after ischemia.

High fatty oxidation (FAO) rates contribute to ischemia-reperfusion injury of the myocardium. Because peroxisome proliferator-activated receptor (PPAR)alpha regulates transcription of several FAO enzymes in the heart, we examined the response of mice with cardiac-restricted overexpression of PPARalpha (MHC-PPARalpha) or whole body PPARalpha deletion including the heart (PPARalpha-/-) to myocardial ischemia-reperfusion injury. Isolated working hearts from MHC-PPARalpha and nontransgenic (NTG) littermates were subjected to no-flow global ischemia followed by reperfusion. MHC-PPARalpha hearts had significantly higher FAO rates during aerobic and postischemic reperfusion (aerobic 1,479 +/- 171 vs. 699 +/- 117, reperfusion 1,062 +/- 214 vs. 601 +/- 70 nmol x g dry wt(-1) x min(-1); P < 0.05) and significantly lower glucose oxidation rates compared with NTG hearts (aerobic 225 +/- 36 vs. 1,563 +/- 165, reperfusion 402 +/- 54 vs. 1,758 +/- 165 nmol x g dry wt(-1) x min(-1); P < 0.05). In hearts from PPARalpha-/- mice, FAO was significantly lower during aerobic and reperfusion (aerobic 235 +/- 36 vs. 442 +/- 75, reperfusion 205 +/- 25 vs. 346 +/- 38 nmol x g dry wt(-1) x min(-1); P < 0.05) whereas glucose oxidation was significantly higher compared with wild-type (WT) hearts (aerobic 2,491 +/- 631 vs. 901 +/- 119, reperfusion 2,690 +/- 562 vs. 1,315 +/- 172 nmol x g dry wt(-1) x min(-1); P < 0.05). Increased FAO rates in MHC-PPARalpha hearts were associated with a markedly lower recovery of cardiac power (45 +/- 9% vs. 71 +/- 6% of preischemic levels in NTG hearts; P < 0.05). In contrast, the percent recovery of cardiac power of PPARalpha-/- hearts was not significantly different from that of WT hearts (80 +/- 8% vs. 75 +/- 9%). This study demonstrates that chronic activation of PPARalpha is detrimental to the cardiac recovery during reperfusion after ischemia.

Keyword: glycolysis

Effects of maternal diabetes on embryogenesis.

Through the use of the in vitro technique of mammalian whole embryo culture, the factors and mechanisms responsible for the increased incidence of congenital malformations among infants of diabetic mothers have been investigated. During early stages of embryogenesis, serum from streptozotocin-induced diabetic rats, hyperglycemia, hypoglycemia, hyperketonemia (beta-hydroxybutyrate), and low molecular weight somatomedin inhibitors are teratogenic or growth inhibitory, or both. Furthermore, combinations of these factors interact to increase the risk of a malformation occurring. In contrast, other factors, such as hyperinsulinemia and excess leucine, , and acetoacetate, have little or no teratogenic potential. Mechanisms of action of the factors are varied and may include alterations of arachidonic metabolism (hyperglycemia), (hypoglycemia), DNA synthesis (beta-hydroxybutyrate), and embryonic nutrition (somatomedin inhibitors). The results demonstrate that the origin of the diabetic embryopathy is multifactorial, that many of the congenital defects are induced early in gestation before the diabetic woman may realize she is pregnant, and that insulin therapy reduces the risk.

Keyword: glycolysis

Evidence of separate pathways for lactate uptake and release by the perfused rat heart.

The simultaneous release and uptake of lactate by the heart has been observed both in vivo and ex vivo; however, the pathways underlying these observations have not been satisfactorily explained. Consequently, the purpose of this study was to test the hypothesis that hearts release lactate from while simultaneously taking up exogenous lactate. Therefore, we determined the effects of fatty acids and diabetes on the regulation of lactate uptake and release. Hearts from control and 1-wk diabetic animals were perfused with 5 mM glucose, 0.5 mM [3-(13)C]lactate, and 0, 0.1, 0.32, or 1.0 mM palmitate. Parameters measured include perfusate lactate concentrations, fractional enrichment, and coronary flow rates, which enabled the simultaneous, but independent, measurements of the rates of 1) uptake of exogenous [(13)C]lactate and 2) efflux of unlabeled lactate from metabolism of glucose. Although the rates of lactate uptake and efflux were both similarly inhibited by the addition of palmitate, (i.e., the ratio of lactate uptake to efflux remained constant), the ratio of lactate uptake to efflux was significantly higher in the controls compared with the diabetic group (1.00 +/- 0.14 vs. 0.50 +/- 0.07, P < 0.002). These data, combined with heterogeneous (13)C enrichment of tissue lactate, pyruvate, and alanine, suggest that glycolytically derived lactate production and oxidation of exogenous lactate operate as functionally separate metabolic pathways. These results are consistent with the concept of an intracellular lactate shuttle.

Keyword: glycolysis

Glucose metabolism in perfused mouse hearts overexpressing human GLUT-4 glucose transporter.

Glucose and fatty metabolism was assessed in isolated working hearts from control C57BL/KsJ-m+/+db mice and transgenic mice overexpressing the human GLUT-4 glucose transporter (db/+-hGLUT-4). Heart rate, coronary flow, cardiac output, and cardiac power did not differ between control hearts and hearts overexpressing GLUT-4. Hearts overexpressing GLUT-4 had significantly higher rates of glucose uptake and and higher levels of glycogen after perfusion than control hearts, but rates of glucose and palmitate oxidation were not different. Insulin (1 mU/ml) significantly increased glycogen levels in both groups. Insulin increased in control hearts but not in GLUT-4 hearts, whereas glucose oxidation was increased by insulin in both groups. Therefore, GLUT-4 overexpression increases , but not glucose oxidation, in the heart. Although control hearts responded to insulin with increased rates of , the enhanced entry of glucose in the GLUT-4 hearts was already sufficient to maximally activate under basal conditions such that insulin could not further stimulate the glycolytic rate.

Keyword: glycolysis

De novo lipogenesis in Atlantic salmon adipocytes.

Carnivorous teleost fish utilize glucose poorly, and the reason for this is not known. It is possible that the capacity of adipocytes to synthesize lipids from carbohydrate precursors through a process known as "de novo lipogenesis" (DNL) is one of the factors that contributes to glucose intolerance in Atlantic salmon.Primary adipocytes from Atlantic salmon differentiated in vitro were incubated with radiolabelled glucose in order to explore the capacity of salmon adipocytes to synthesize and deposit lipids from glucose through DNL. The lipid-storage capacity of adipocytes incubated with glucose was compared with that of cells incubated with the fatty . Quantitative PCR and immunohistochemistry were used to assess changes of genes and proteins involved in glucose and lipid transport and metabolism.Less than 0.1% of the radiolabelled glucose was metabolized to the fatty acids 16:0 and the stearoyl-CoA desaturase products 16:1 and 18:1 by DNL, whereas approximately 40% was converted to glycerol to form the triacylglycerol backbone of lipids. Transcriptional analysis indicated that adipocytes ensure the availability of necessary cofactors and other substrates for lipid synthesis and storage from , the pentose phosphate pathway and glyceroneogenesis.We have shown for the first time that the DNL pathway is active in fish adipocytes. The capacity of the pathway to convert glucose into cellular lipids for storage is relatively low.The limited capacity of adipocytes to utilize glucose as a substrate for lipid deposition may contribute to glucose intolerance in salmonids.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: glycolysis

Differences in fuel utilization between trout and human thrombocytes in physiological media.

Cell culture preparations now play a significant and essential role in physiological and biochemical studies of cell biology. However, the fuels offered in cell culture media are only glucose and glutamine, plus whatever might be in the added sera. It is currently difficult to find a rational way forward on this problem, as there are few data on what fuels cells use in vivo or even in an in vitro physiological situation. A recent study on human platelets redressed the situation somewhat by finding that 75% of ATP turnover could be accounted for by aerobic , and by the oxidation of glucose, hydroxybutyrate, acetate, glutamine, palmitate and oleate. In the present study we used a similar strategy to investigate fuel choices by trout thymocytes, cells with a similar function but from a different phylogenetic group. When these cells were presented with a physiological medium, we found that aerobic accounted for 9% of total ATP turnover, glucose and glutamine oxidation made a combined contribution of 2.3%, oleate and palmitate oxidation accounted for 15%, and 74% was unaccounted for. These patterns of fuel use are very different from that in human platelets. They demonstrate the cell- and animal-specific nature of cellular metabolism and again expose the inadequacy of the fuel component in culture media.

Keyword: glycolysis

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, , oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: glycolysis

Role of fatty acids in the recovery of cardiac function during resuscitation from hemorrhagic shock.

This study tested the hypothesis that removal of fatty acids as a fuel source would improve cardiac efficiency at the expense of reduced cardiac contractile function in the isolated working heart after hemorrhage-retransfusion. Non-heparinized male Sprague-Dawley rats were anesthetized with ketamine-xylazine and were hemorrhaged to a mean arterial blood pressure of 40 mmHg for 1 h. Two-thirds volume of shed blood was reinfused together with 0.9% NaCl in a volume equal to 2.3 times the shed blood volume, followed by continuous infusion of 0.9% NaCl at 10 mL/kg per h for 3 h. Hearts were removed and perfused in closed, recirculating working mode for 60 min to measure hydraulic work and cardiac efficiency. Rates of and glucose oxidation were assessed with [5-3H/U-14C] glucose (11 mM) in the absence or presence of 0.4 mM palmitate. Compared to baseline measurements, hemorrhage-retransfusion significantly reduced arterial blood glucose (228+/-7 versus 118+/-12 mg/dL) and non-esterified fatty concentrations (0.36+/-0.01 versus 0.30+/-0.02 mM), while elevating blood lactate (0.8+/-0.1 versus 2.5+/-0.4 mM). Perfusion of sham hearts with glucose-only did not alter cardiac work compared to shams perfused with glucose plus palmitate. However, shocked hearts perfused with glucose-only demonstrated a significant reduction in cardiac work compared to shocked hearts perfused with glucose plus palmitate and compared to sham hearts perfused with glucose only (P < 0.05, repeated measures ANOVA). Shocked hearts perfused with glucose plus palmitate showed no reduction in cardiac work compared to shams. Shocked hearts perfused with glucose-only had increased glucose oxidation rates compared to shams perfused with glucose plus palmitate. In sham hearts perfused with glucose-only, myocardial glycogen and triacylglycerol contents were significantly reduced compared to hearts freeze-clamped in situ. These endogenous fuels were not decreased in shocked hearts. These data indicate that hemorrhagic shock renders the heart unable to mobilize endogenous fuels, and suggest that withdrawal of fatty oxidation will impair myocardial energy metabolism during resuscitation.

Keyword: glycolysis

Hyperlipidaemia is associated with increased insulin-mediated glucose metabolism, reduced fatty metabolism and normal blood pressure in transgenic mice overexpressing human apolipoprotein C1.

Insulin resistance for glucose metabolism is associated with hyperlipidaemia and high blood pressure. In this study we investigated the effect of primary hyperlipidaemia on basal and insulin-mediated glucose and on non-esterified fatty (NEFA) metabolism and mean arterial pressure in hyperlipidaemic transgenic mice overexpressing apolipoprotein C1 (APOC1). Previous studies have shown that APOC1 transgenic mice develop hyperlipidaemia primarily because of an impaired hepatic uptake of very low density lipoprotein (VLDL).Basal and hyperinsulinaemic (6 mU.kg-1.min-1), euglycaemic (7 mmol/l) clamps with 3(-)3H-glucose or 9,10(-)3H- infusions and in situ freeze clamped tissue collection were carried out.The APOC1 mice showed increased basal plasma cholesterol, triglyceride, NEFA and decreased glucose concentrations compared with wild-type mice (7.0 +/- 1.2 vs 1.6 +/- 0.1, 9.1 +/- 2.3 vs 0.6 +/- 0.1, 1.9 +/- 0.2 vs 0.9 +/- 0.1 and 7.0 +/- 1.0 vs 10.0 +/- 1.1 mmol/l, respectively, p < 0.05). Basal whole body glucose clearance was increased twofold in APOC1 mice compared with wild-type mice (18 +/- 2 vs 10 +/- 1 ml.kg-1.min-1, p < 0.05). Insulin-mediated whole body glucose uptake, (generation of 3H2O) and glucose storage increased in APOC1 mice compared with wild-type mice (339 +/- 28 vs 200 +/- 11; 183 +/- 39 vs 128 +/- 17 and 156 +/- 44 vs 72 +/- 17 mumol.kg-1.min-1, p < 0.05, respectively), corresponding with a twofold to threefold increase in skeletal muscle glycogenesis and de novo lipogenesis from 3-(3)H-glucose in skeletal muscle and adipose tissue (p < 0.05). Basal whole body NEFA clearance was decreased threefold in APOC1 mice compared with wild-type mice (98 +/- 21 vs 314 +/- 88 ml.kg-1.min-1, p < 0.05). Insulin-mediated whole body NEFA uptake, NEFA oxidation (generation of 3H2O) and NEFA storage were lower in APOC1 mice than in wild-type mice (15 +/- 3 vs 33 +/- 6; 3 +/- 2 vs 11 +/- 4 and 12 +/- 2 vs 22 +/- 4 mumol.kg-1.min-1, p < 0.05) in the face of higher plasma NEFA concentrations (1.3 +/- 0.3 vs 0.5 +/- 0.1 mmol/l, p < 0.05), respectively. Mean arterial pressure and heart rate were similar in APOC1 vs wild-type mice (82 +/- 4 vs 85 +/- 3 mm Hg and 459 +/- 14 vs 484 +/- 11 beats.min-1).1) Hyperlipidaemic APOC1 mice show reduced NEFA and increased glucose metabolism under both basal and insulin-mediated conditions, suggesting an intrinsic defect in NEFA metabolism. Primary hyperlipidaemia alone in APOC1 mice does not lead to insulin resistance for glucose metabolism and high blood pressure.

Keyword: glycolysis

Differences in myocardial ischemic tolerance between 1- and 7-day-old rabbits.

Between 1 and 7 days of life, the newborn rabbit heart shifts from predominantly using carbohydrates to predominantly using fatty acids as an energy substrate. We therefore used isolated working hearts from 1- or 7-day-old rabbits to determine the effects of fatty acids on myocardial glucose use and the ability of hearts to recover following various periods of transient no-flow ischemia. One-day-old hearts were perfused via the inferior vena cava and ejected buffer through the cannulated aorta and pulmonary artery. Seven-day-old hearts were perfused via the left atrium and ejected buffer through the cannulated aorta. To measure glucose use, hearts were perfused with 11 mM [3H, 14C]glucose, 3% albumin, and 500 microU insulin/mL, in the presence or absence of 0.4 mM palmitate. In the absence of fatty acids, glycolytic rates were similar in 1- and 7-day-old hearts, whereas glucose oxidation rates were 5 times greater in 7-day-old hearts. Palmitate did not have any major effects on overall glucose use in 1-day-old hearts, but did markedly inhibit and glucose oxidation in 7-day-old hearts. A series of hearts were also subjected to periods (25-60 min) of no-flow ischemia, followed by 30 min of aerobic reperfusion. In the absence of palmitate, 1-day-old hearts subjected to ischemic periods of up to 60 min recovered some degree of mechanical function during reperfusion, whereas 7-day-old rabbit hearts failed to recover if hearts were subjected to ischemic periods of 35 min or longer.(ABSTRACT TRUNCATED AT 250 WORDS).

Keyword: glycolysis

Effects of fatty oxidation on glucose utilization by isolated hepatocytes.

We have studied the inhibitory action of long- and short-chain fatty acids on hepatic glucose utilization in hepatocytes isolated from fasted rats. The rates of hepatic glucose phosphorylation and were determined from the tritiated products of [2-3H] and [6-3H]glucose metabolism, respectively. The difference between these was taken as an estimate of the \'cycling\' between glucose and glucose-6-phosphate. In the presence of 40 mM glucose this cycling was estimated at 0.68 mumol/min/g wet wt. Glucose phosphorylation was unaffected during palmitate and hexanoate oxidation to ketone bodies but was inhibited. The rate of glucose cycling was increased during this phase to 1.25 mumol/min/g. Following the complete metabolism of the fatty acids, was reinstated and cycling rates returned to control levels. Hepatic glucose cycling appears to be an important component of the glucose/fatty cycle.

Keyword: glycolysis

Metabolomic and transcriptomic responses induced in the livers of pigs by the long-term intake of resistant starch.

The present study investigated metabolomic and transcriptomic responses in the livers of pigs to evaluate the effects of resistant starch on the body\'s metabolism at the extraintestinal level. Thirty-six Duroc× Landrace × Large White growing barrows (70 d of age) were randomly allocated to either the corn starch (CS) group or the raw potato starch (RPS) group with a randomized complete block design; each group consisted of 6 replicates (pens), with 3 pigs per pen. Pigs in the CS group were offered a corn-soybean-based diet, whereas pigs in the RPS group were put on a diet in which 230 (growing) or 280 g/kg (finishing) purified CS was replaced with purified RPS during a 100-d trial. The livers of pigs were collected for metabolome and gene expression analysis. Gas chromatography-mass spectrometry analysis showed that compared with the CS diet, the RPS diet decreased ( < 0.05) cholesterol and as well as increased ( < 0.05) 3-hydroxybutyric , which indicated the reduction of adipose weight and fatty biosynthesis and the elevation of fatty β-oxidation. In addition, 2-ketoglutaric and glucose-6-phosphate were increased (< 0.05) although pyruvic was decreased ( < 0.05) in the RPS group, indicating the upregulated capacity of glucose phosphorylation and glycolysis. Microarray analysis showed that the mRNA expression of (), (), and () were downregulated ( < 0.05) whereas (), (), and () were upregulated ( < 0.05) in the RPS diet, indicating a decrease in fatty intake and synthesis and an increase in fatty oxidation and glycerophospholipid synthesis. The results demonstrated that the long-term consumption of RPS could modulate hepatic lipid metabolism by decreasing fatty synthesis as well as increasing lipid oxidation and glycerophospholipid synthesis.

Keyword: glycolysis

Insulin regulation of fatty synthase gene transcription: roles of USF and SREBP-1c.

The transcriptional regulation of lipogenesis is a highly coordinated process occurring in concert with transcriptional as well as post-transcriptional regulation of enzymes involved in and gluconeogenesis. Fatty synthase (FAS) plays a central role in de novo lipogenesis by converting acetyl-CoA and malonyl-CoA into the final end product, palmitate, which can subsequently be esterified into triacylglycerols and then stored in adipose tissue. Ultimately, this helps to prevent buildup of excess glucose in other types of cells and tissues, the effects of which can be readily observed in the pathophysiology of disease states such as Type-11 diabetes and obesity. Thus, elucidating the transcriptional mechanisms of lipogenic enzyme genes is important for understanding the normal regulation of lipogenesis and ultimately the dysregulation that may occur in certain metabolic disease. In this review, we discuss advances in our understanding of the regulation of lipogenesis at the genetic level, with a special emphasis on the common cis- and trans-acting factors involved in regulation of FAS. Two transcription factors, Upstream Stimulatory Factor (USF) and Sterol Regulatory Element Binding Protein-lc (SREBP-lc), seem to play a dominant and possibly cooperative role in regulating FAS transcription.

Keyword: glycolysis

Substrate utilization of adult cardiac myocytes.

Cultured adult myocytes are in a state of basal metabolism. When glucose is the only exogenous substrate, they produce lactate over CO2 at a constant rate of 2.7 from this substrate. Increase of oxygen tension does not change this behaviour. Insulin preferentially increases lactate formation, dichloroacetate only CO2 production. The fact that the lactate/CO2 ratio can be varied from 0.5 to 16 indicates that there is no close coupling between glycolytic flux and pyruvate oxidation. Both exogenous lactate and fatty acids are used preferentially over glucose. But increase of fatty oxidation and inhibition of glucose oxidation are not complementary. Glycolytic flux is only slightly decreased when fatty oxidation is already saturated. The results indicate that fatty acids interact with glucose oxidation primarily by inactivation of the pyruvate dehydrogenase. Neither insulin nor dichloroacetate in the presence of glucose inhibit fatty oxidation.

Keyword: glycolysis

Short-term cigarette smoke exposure leads to metabolic alterations in lung alveolar cells.

Cigarette smoke (CS)-induced alveolar destruction and energy metabolism changes are known contributors to the pathophysiology of chronic obstructive pulmonary disease (COPD). This study examines the effect of CS exposure on metabolism in alveolar type II cells. Male A/J mice (8 wk old) were exposed to CS generated from a smoking machine for 4 or 8 weeks, and a recovery group was exposed to CS for 8 weeks and allowed to recover for 2 weeks. Alveolar type II cells were isolated from air- or CS- exposed mice. Acute CS exposure led to a reversible airspace enlargement in A/J mice as measured by the increase in mean linear intercept, indicative of alveolar destruction. The effect of CS exposure on cellular respiration was studied using the XF Extracellular Flux Analyzer. A decrease in respiration while metabolizing glucose was observed in the CS-exposed group, indicating altered that was compensated by an increase in palmitate utilization; palmitate utilization was accompanied by an increase in the expression of CD36 and carnitine-palmitoyl transferase 1 in type II alveolar cells for the transport of palmitate into the cells and into mitochondria, respectively. The increase in palmitate use for energy production likely affects the surfactant biosynthesis pathway, as evidenced by the decrease in phosphatidylcholine levels and the increase in phospholipase A2 activity after CS exposure. These findings help our understanding of the mechanism underlying the surfactant deficiency observed in smokers and provide a target to delay the onset of COPD.

Keyword: glycolysis

Skeletal muscle fibers in suspension: a new approach to the study of oxidative and glycolytic metabolism in differentiated muscle.

A preparation of suspended fibers from m. flexor digitorum brevis of the rat was characterized with respect to morphological features, and its relevance for the study of muscular metabolism investigated. The activities of oxidative (palmitate and pyruvate oxidation) and glycolytic (lactate formation) pathways were enhanced in myofiber suspensions when compared to intact whole muscle. The rate of was stimulated about two-fold by insulin in both the myofiber suspensions and intact muscle. Parameters of oxidative metabolism responded similarly to metabolic effectors in the myofiber suspensions and in intact muscle. It is concluded that the myofiber suspensions have distinct advantages over intact muscle for biochemical and pharmacological studies.

Keyword: glycolysis

Global metabolic profiling of infection by an oncogenic virus: KSHV induces and requires lipogenesis for survival of latent infection.

Like cancer cells, virally infected cells have dramatically altered metabolic requirements. We analyzed global metabolic changes induced by latent infection with an oncogenic virus, Kaposi\'s Sarcoma-associated herpesvirus (KSHV). KSHV is the etiologic agent of Kaposi\'s Sarcoma (KS), the most common tumor of AIDS patients. Approximately one-third of the nearly 200 measured metabolites were altered following latent infection of endothelial cells by KSHV, including many metabolites of anabolic pathways common to most cancer cells. KSHV induced pathways that are commonly altered in cancer cells including , the pentose phosphate pathway, amino production and fatty synthesis. Interestingly, over half of the detectable long chain fatty acids detected in our screen were significantly increased by latent KSHV infection. KSHV infection leads to the elevation of metabolites involved in the synthesis of fatty acids, not degradation from phospholipids, and leads to increased lipid droplet organelle formation in the infected cells. Fatty synthesis is required for the survival of latently infected endothelial cells, as inhibition of key enzymes in this pathway led to apoptosis of infected cells. Addition of to latently infected cells treated with a fatty synthesis inhibitor protected the cells from death indicating that the products of this pathway are essential. Our metabolomic analysis of KSHV-infected cells provides insight as to how oncogenic viruses can induce metabolic alterations common to cancer cells. Furthermore, this analysis raises the possibility that metabolic pathways may provide novel therapeutic targets for the inhibition of latent KSHV infection and ultimately KS tumors.

Keyword: glycolysis

Lipid metabolism in cancer cachexia.

The effect of cancer cachexia on the oxidative metabolism of lipids has been studied in mice transplanted either with the MAC16 adenocarcinoma, which induces profound loss of body weight and depletion of lipid stores, or the MAC13 adenocarcinoma, which is the same histological type, but which grows without an effect on host body weight or lipid stores. While oxidation of D-[U-14C]glucose did not differ between animals bearing tumours of either type and non-tumour bearing controls, oxidation of [1-14C]triolein administered by intragastric intubation was significantly (P less than 0.05) higher in animals bearing the MAC16 tumour than in either non tumour-bearing controls or in animals bearing the MAC13 tumour. Intestinal absorption of [14C]lipid was significantly (P less than 0.05) reduced in animals bearing the MAC13 tumour when compared with either non tumour-bearing animals or MAC16 tumour-bearing animals, but was not significantly different in the latter two groups. The level of labelled lipids in heart and adipose tissue after an oral [14C]lipid load was significantly lower in animals bearing the MAC16 tumour compared with the other two groups. The level of tumour lipids was also higher in the MAC16 than in the MAC13 tumour after both an oral [14C]lipid load or by direct injection of [U-14C]palmitate complexed to albumin into epididymal fat pads. Oxidation of [U-14C]palmitate was also significantly enhanced in liver and heart homogenates from animals bearing the MAC16 tumour. These results suggest that in cachectic tumour-bearing animals mobilisation of body lipids is accompanied by an increased utilisation.

Keyword: glycolysis

A stimulatory effect of FFA on unmasked in cells with impaired oxidative capacity.

The physiological importance of the glucose fatty cycle has been controversial. Many studies have failed to demonstrate an inhibitory effect of free fatty acids (FFA) on glucose utilization. Using both hepatoma cells (Hep G2) and human erythrocytes, which have poor oxidative capacity and metabolize glucose primarily anaerobically, we have demonstrated a unique stimulatory effect of FFA on . Fructose 2,6-bisphosphate (F-2,6-P2) concentrations also increased significantly in Hep G2 cells incubated with . In contrast, F-2,6-P2 concentrations fell in primary cultured hepatocytes incubated with in association with increased oxidation of FFA and accumulation of beta-hydroxybutyrate. We propose that a stimulatory effect of FFA on reported here for the first time may have been masked in previous studies performed in tissues in which the oxidation of FFA and the accumulation of intermediates such as citrate may have decreased F-2,6-P2 concentrations. We conclude that the spectrum of FFA effects in probably depends on tissue oxidative capacity.

Keyword: glycolysis

Fatty synthase activity in tumor cells.

While normal tissues are tightly regulated by nutrition and a carefully balanced system of and fatty synthesis, tumor cells are under significant evolutionary pressure to bypass many of the checks and balances afforded normally. Cancer cells have high energy expenditure from heightened proliferation and metabolism and often show increased lipogenesis. Fatty synthase (FASN), the enzyme responsible for catalyzing the ultimate steps of fatty synthesis in cells, is expressed at high levels in tumor cells and is mostly absent in corresponding normal cells. Because of the unique expression profile of FASN, there is considerable interest not only in understanding its contribution to tumor cell growth and proliferation, but also in developing inhibitors that target FASN specifically as an anti-tumor modality. Pharmacological blockade of FASN activity has identified a pleiotropic role for FASN in mediating aspects of proliferation, growth and survival. As a result, a clearer understanding of the role of FASN in tumor cells has been developed.

Keyword: glycolysis

Fatty and glucose oxidation by cultured rat heart cells.

Keyword: glycolysis

Metabolic studies on isolated hair follicles: hair follicles engage in aerobic and do not demonstrate the glucose fatty cycle.

The matrix cells of the hair follicle have one of the highest rates of cell division in the mammalian body, but their fuel metabolism is poorly understood, due mainly to the difficulty in obtaining viable intact follicles from the skin. We have previously shown that viable and intact rat hair follicles can be isolated by shearing, and in this study we now report on their fuel metabolism. In this study we have shown that the hair follicle exhibits aerobic , in that of the total glucose utilized by the hair follicle, only 10% is oxidized to CO2. We have also shown that, in the absence of glucose, the hair follicle is capable of utilizing other fuels such as palmitate and beta-hydroxybutyrate. However, neither palmitate or beta-hydroxybutyrate had any effect on the rate of glucose utilization or on [U-14C] glucose oxidation, showing that glucose sparing via the glucose fatty cycle does not operate in the hair follicle. Measurements of glucose flux through the pentose phosphate pathway accounted for only 3% of the total glucose utilized by the hair follicle, although this value represented 32% of the total glucose oxidized. Both palmitate and beta-hydroxybutyrate inhibited glucose flux through the pentose phosphate pathway.

Keyword: glycolysis

Glucose and carbachol generate 1,2-diacylglycerols by different mechanisms in pancreatic islets.

Diacylglycerols (DAG) modulate secretory responses by the activation of protein kinase C. Early changes in DAG formation induced by the muscarinic receptor agonist carbachol were compared to those caused by the nutrient secretagogue glucose in pancreatic islets. Turnover rates of DAG were investigated in radiolabeling experiments, whereas changes in total mass and fatty composition of DAG were assessed by gas-liquid chromatography. When islet lipids were labeled to steady state in tissue culture with [3H]glycerol, carbachol induced a rapid (10 s) and sustained increase of [3H]DAG generation. In contrast, glucose stimulation failed to increase [3H]glycerol containing DAG, and this was probably due to the isotopic dilution of the label secondary to enhanced . This was substantiated by following the transfer of 14C from glucose into DAG. Within 1 min of acute exposure of islets to D-[U-14C]-glucose at stimulatory concentrations, DAG labeling increased fivefold representing up to 2% of total glucose usage. Similar stimulation of 14C incorporation into other neutral lipids and inositol phospholipids was observed, suggesting the enhanced de novo synthesis of phosphatidic , the common precursor for DAG, and inositol phospholipids from glycolytic intermediates. Transfer of 14C from glucose was not stimulated by agents such as carbachol and exogenous phospholipase C that act primarily on inositol phospholipid breakdown. The total mass of islet DAG was increased by 60% after both carbachol and glucose stimulation. However, analysis of the fatty composition of carbachol-generated DAG revealed at the early time point (10 s) a prevalent stearoyl-arachidonoyl configuration similar to that reported for inositol phospholipids. This pattern shifted to a DAG enriched in at a later time point. Glucose-stimulated islets displayed a predominance of containing DAG, indicating increased de novo synthesis of the putative second messenger rather than its formation by inositol phospholipid hydrolysis. Indeed, steady-state labeling of these phospholipids with [3H]inositol confirmed this idea since only carbachol caused detectable inositol phospholipid hydrolysis. Thus, although protein kinase C may be activated by both carbachol and glucose, the two secretagogues generate diacylglycerols through different mechanisms.

Keyword: glycolysis

Changes in beta-oxidation and related enzymes during the life cycle of Strongyloides ratti (Nematoda).

Keyword: glycolysis

Integration of proteomics and metabolomics data of early and middle season Hass avocados under heat treatment.

Ripening heterogeneity of Hass avocados results in inconsistent quality fruit delivered to the triggered and ready to eat markets. This research aimed to understand the effect of a heat shock (HS) prior to controlled atmosphere (CA) storage on the reduction of ripening heterogeneity. HS prior to CA storage reduces more drastically the ripening heterogeneity in middle season fruit. Via correlation network analysis we show the different metabolomics networks between HS and CA. High throughput proteomics revealed 135 differentially expressed proteins unique to middle season fruit triggered by HS. Further integration of metabolomics and proteomics data revealed that HS reduced the glycolytic throughput and induced protein degradation to deliver energy for the alternative ripening pathways. l-isoleucine, l-valine, l-aspartic and ubiquitin carboxyl-terminal hydrolase involved in protein degradation were positively correlated to HS samples. Our study provides new insights into the effectiveness of HS in synchronizing ripening of Hass avocados.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: glycolysis

AIF loss deregulates hematopoiesis and reveals different adaptive metabolic responses in bone marrow cells and thymocytes.

Mitochondrial metabolism is a tightly regulated process that plays a central role throughout the lifespan of hematopoietic cells. Herein, we analyze the consequences of the mitochondrial oxidative phosphorylation (OXPHOS)/metabolism disorder associated with the cell-specific hematopoietic ablation of apoptosis-inducing factor (AIF). AIF-null (AIF ) mice developed pancytopenia that was associated with hypocellular bone marrow (BM) and thymus atrophy. Although myeloid cells were relatively spared, the B-cell and erythroid lineages were altered with increased frequencies of precursor B cells, pro-erythroblasts I, and basophilic erythroblasts II. T-cell populations were dramatically reduced with a thymopoiesis blockade at a double negative (DN) immature state, with DN1 accumulation and delayed DN2/DN3 and DN3/DN4 transitions. In BM cells, the OXPHOS/metabolism dysfunction provoked by the loss of AIF was counterbalanced by the augmentation of the mitochondrial biogenesis and a shift towards anaerobic . Nevertheless, in a caspase-independent process, the resulting excess of reactive oxygen species compromised the viability of the hematopoietic stem cells (HSC) and progenitors. This led to the progressive exhaustion of the HSC pool, a reduced capacity of the BM progenitors to differentiate into colonies in methylcellulose assays, and the absence of cell-autonomous HSC repopulating potential in vivo. In contrast to BM cells, AIF thymocytes compensated for the OXPHOS breakdown by enhancing fatty β-oxidation. By over-expressing CPT1, ACADL and PDK4, three key enzymes facilitating fatty β-oxidation (e.g., assimilation), the AIF thymocytes retrieved the ATP levels of the AIF cells. As a consequence, it was possible to significantly reestablish AIF thymopoiesis in vivo by feeding the animals with a high-fat diet complemented with an antioxidant. Overall, our data reveal that the mitochondrial signals regulated by AIF are critical to hematopoietic decision-making. Emerging as a link between mitochondrial metabolism and hematopoietic cell fate, AIF-mediated OXPHOS regulation represents a target for the development of new immunomodulatory therapeutics.

Keyword: glycolysis

Increase in fatty oxidation in calvaria cells cultured with diphosphonates.

1. Cultured calvaria cells oxidized palmitate and octanoate to CO2 and water-soluble products. 2. When these cells were treated for 6 days with 0.025 and 0.25 mM-dichloromethanediphosphonate, oxidation of palmitate was increased, whereas that of octanoate was influenced less. 3. When the rate of oxidation was raised by increasing the palmitate concentration in the medium, the effect of the diphosphonate was decreased and finally disappeared. 4. 1-Hydroxyethane-1,1-diphosphonate had only minor effects. 5. The increase in palmitate oxidation appeared 2 days after the addition of dichloromethanediphosphonate, simultaneously with a fall in lactate production. (Inhibition of by diphosphonates has already been shown.) 6. Cycloheximide, an inhibitor of protein synthesis, did not influence the effect of dichloromethanediphosphonate on the oxidation of palmitate and the production of lactate. 7. Cells cultured with dichloromethanediphosphonate showed a faster uptake of than did control cells. However, this observation did not explain the increased palmitate oxidation, since uptake was much faster than oxidation, and was therefore not the rate-limiting step. 8. 2-Bromopalmitate, an inhibitor of fatty oxidation, did not influence the inhibition of by the diphosphonates. This inhibition, therefore, did not result from the increased oxidation of palmitate. It is also unlikely that the increased oxidation of palmitate is connected with the inhibition of .

Keyword: glycolysis

THE EFFECTS OF QUINIDINE ON THE METABOLISM OF GLUCOSE-U-C14 AND PALMITATE-U-C14 BY HEART MUSCLE.

Keyword: glycolysis

The effect of emetine on metabolism and contractility of the isolated rat heart.

Keyword: glycolysis

Stages in the incorporation of fatty acids into red blood cells.

Mature human erythrocytes were incubated with (14)C-labeled bound to crystalline human albumin. Energy-dependent incorporation of the labeled into cell membrane phospholipids occurred, and various stages in this incorporation were defined. Initially the was rapidy transferred from the albumin to a "superficial" membrane pool of free fatty (F-1), which was removable when the cells were washed with defatted albumin. This process was independent of red cell metabolism. The labeled fatty then passed into a second "deeper" membrane pool of free fatty acids (F-2), which was not extractable with albumin. This process was energy-dependent and proceeded at a slower rate than the initial transfer from albumin to F-1. Ultimately the labeled fatty was incorporated into phosphatides (PL). This process also was dependent upon cellular metabolism. The kinetics of pulse label studies suggest that the processes observed were sequential and that precursor-product relationships exist between the F-1 and F-2 pools and the F-2 and PL pools. [Formula: see text] From the size and specific activities of these pools, calculations of the extent of phospholipid turnover were made. An approximate figure of 2% /hr or 30 nmoles/ml of packed red blood cells per hr was obtained. The figure was further calculated to represent an energy cost to the red blood cell of approximately 5% of the energy available from .

Keyword: glycolysis

Fatty acids alter glycerolipid metabolism and induce lipid droplet formation, syncytialisation and cytokine production in human trophoblasts with minimal glucose effect or interaction.

The diabetic pregnancy is characterized by maternal hyperglycaemia and dyslipidaemia, such that placental trophoblast cells are exposed to both. The objective was to determine the effects of hyperglycaemia, elevated non-esterified fatty acids (NEFA) and their interactions on trophoblast cell metabolism and function. Trophoblasts were isolated from normal term human placentas and established in culture for 16 h prior to experiments. Glucose utilisation, fatty oxidation and fatty esterification were determined using radiolabelled metabolic tracer methodology at various glucose and NEFA concentrations. Trophoblast lipid droplet formation including adipophilin mRNA expression, viability, apoptosis, syncytialisation, secretion of hormones and pro-inflammatory cytokines were also assessed. Glucose utilisation via was near maximal at the low physiological glucose concentration of 4mM; whereas NEFA esterification into triacylglycerol and diacylglycerol increased linearly with increasing NEFA concentrations without evidence of plateau. Culture of trophoblasts in 0.25 mM NEFA for 24h upregulated fatty esterification processes, inhibited fatty oxidation, inhibited glycerol release (a marker of lipolysis) and promoted adipophilin and lipid droplet formation, all consistent with upregulation of fatty storage and buffering capacity. NEFA also promoted trophoblast syncytialisation and TNFalpha, IL-1beta, IL-6 and IL-10 production without effects on cell viability, apoptosis or hormone secretion. Hyperglycaemia caused intracellular glycogen accumulation and reduced lipid droplet formation, but had no other effects on trophoblast metabolism or function. NEFA have effects on trophoblast metabolism and function, mostly independent of glucose, that may have protective as well as pathophysiological roles in pregnancies complicated by diabetes and/or obesity.Copyright 2009 Elsevier Ltd. All rights reserved.

Keyword: glycolysis

Metabolomic and lipidomic analysis of serum from mice exposed to an internal emitter, cesium-137, using a shotgun LC-MS(E) approach.

In this study ultra performance liquid chromatography (UPLC) coupled to time-of-flight mass spectrometry in the MS(E) mode was used for rapid and comprehensive analysis of metabolites in the serum of mice exposed to internal exposure by Cesium-137 ((137)Cs). The effects of exposure to (137)Cs were studied at several time points after injection of (137)CsCl in mice. Over 1800 spectral features were detected in the serum of mice in positive and negative electrospray ionization modes combined. Detailed statistical analysis revealed that several metabolites associated with amino metabolism, fatty metabolism, and the TCA cycle were significantly perturbed in the serum of (137)Cs-exposed mice compared with that of control mice. While metabolites associated with the TCA cycle and increased in their serum abundances, fatty acids such as linoleic and were detected at lower levels in serum after (137)Cs exposure. Furthermore, phosphatidylcholines (PCs) were among the most perturbed ions in the serum of (137)Cs-exposed mice. This is the first study on the effects of exposure by an internal emitter in serum using a UPLC-MS(E) approach. The results have put forth a panel of metabolites, which may serve as potential serum markers to (137)Cs exposure.

Keyword: glycolysis

Crabtree effect in tumoral pancreatic islet cells.

The relationship between and respiration was examined in a model of pancreatic B-cell dysfunction, namely in tumoral insulin-producing cells of the RINm5F line. A rise in D-glucose concentration from 2.8 to 16.7 mM increased the utilization of D-[5-3H]glucose and production of [14C]lactate from D-[U-14C]glucose, whereas decreasing the oxidation of either D-[U-14C]glucose or D-[6-14C]glucose. Whereas 2.8 mM D-glucose augmented O2 uptake above basal value, a further rise in D-glucose concentration to 16.7 mM decreased respiration, which remained higher, however, than basal value. Whether at low or high concentration, D-glucose exerted a pronounced sparing action upon the oxidation of endogenous nutrients in cells prelabeled with either L-[U-14C]glutamine or [14C]palmitate and, nevertheless, augmented above basal value the rate of lipogenesis, ATP/ADP content, adenylate charge, and cytosolic NADH/NAD+ and NADPH/NADP+ ratios. The generation of ATP resulting from the catabolism of either exogenous D-glucose or endogenous nutrients was not affected by the rise in hexose concentration from 2.8 to 16.7 mM. Thus, in sharp contrast with the situation found in normal islet cells, a rise in D-glucose concentration, instead of stimulating mitochondrial oxidative events, caused, through a Crabtree effect, inhibition of hexose oxidation and O2 consumption in tumoral islet cells.

Keyword: glycolysis

Energetic response of coronary endothelial cells to hypoxia.

The response of endothelial energy metabolism to oxygen supply was studied in cultured coronary endothelial cells from the rat at defined PO2 levels between 0.1 and 100 Torr. In the presence of glucose (5 mM), endothelial respiration (4 nmol O2.min-1.mg protein-1) was independent of the exterior PO2 greater than 3 Torr; oxygen consumption was half maximal at 0.8 Torr. At 100 Torr, lactate production was 26 nmol.min-1.mg protein-1; the decrease of the PO2 to 0.1 Torr resulted in a 2.2-fold increase in lactate production. The contents of ATP, ADP, and AMP were 21, 4, and 2 nmol/mg protein, respectively; they remained constant for 2.5-h incubations at PO2 levels between 0.1 and 100 Torr. In the presence of palmitate (100 microM) plus glutamine (0.5 mM), oxygen consumption was 8 nmol.min-1.mg protein-1 at PO2 levels greater than 3 Torr, and the half-maximal rate was again observed at 0.8 Torr. Lactate production was negligible. At PO2 levels greater than 3 Torr, the cells remained well energized. Below 3 Torr, however, the adenine nucleotide contents rapidly declined. These results demonstrate that the oxygen demand of coronary endothelial cells is low compared with the beating myocardium. In the presence of glucose, aerobic is pronounced and the Pasteur effect small. In severe hypoxia (PO2 less than 0.1 Torr) the energetic state remained stable. In the absence of glucose, the energetic state of coronary endothelial cells is sensitive to the exterior PO2 less than 3 Torr, declining concomitantly with the decrease in respiration.

Keyword: glycolysis

Insulin acutely improves mitochondrial function of rat and human skeletal muscle by increasing coupling efficiency of oxidative phosphorylation.

Insulin is essential for the regulation of fuel metabolism and triggers the uptake of glucose by skeletal muscle. The imported glucose is either stored or broken down, as insulin stimulates glycogenesis and ATP synthesis. The mechanism by which ATP production is increased is incompletely understood at present and, generally, relatively little functional information is available on the effect of insulin on mitochondrial function. In this paper we have exploited extracellular flux technology to investigate insulin effects on the bioenergetics of rat (L6) and human skeletal muscle myoblasts and myotubes. We demonstrate that a 20-min insulin exposure significantly increases (i) the cell respiratory control ratio, (ii) the coupling efficiency of oxidative phosphorylation, and (iii) the glucose sensitivity of anaerobic . The improvement of mitochondrial function is explained by an insulin-induced immediate decrease of mitochondrial proton leak. Palmitate exposure annuls the beneficial mitochondrial effects of insulin. Our data improve the mechanistic understanding of insulin-stimulated ATP synthesis, and reveal a hitherto undisclosed insulin sensitivity of cellular bioenergetics that suggests a novel way of detecting insulin responsiveness of cells.© 2013.

Keyword: glycolysis

Inhibition of very long chain acyl-CoA dehydrogenase during cardiac ischemia.

The heart utilizes primarily fatty acids for energy production. During ischemia, however, diminished oxygen supply necessitates a switch from beta-oxidation of fatty acids to glucose utilization and . Molecular mechanisms responsible for these alterations in metabolism are not fully understood. Mitochondrial acyl-CoA dehydrogenase catalyzes the first committed step in the beta-oxidation of fatty acids. In the current study, an in vivo rat model of myocardial ischemia was utilized to determine whether specific acyl-CoA dehydrogenases exhibit ischemia-induced alterations in activity, identify mechanisms responsible for changes in enzyme function, and assess the effects on mitochondrial respiration. Very long chain acyl-CoA dehydrogenase (VLCAD) activity declined 34% during 30 min of ischemia. Loss in activity appeared specific to VLCAD as medium chain acyl-CoA dehydrogenase activity remained constant. Loss in VLCAD activity during ischemia was not due to loss in protein content. In addition, activity was restored in the presence of the detergent Triton X-100, suggesting that changes in the interaction between the protein and inner mitochondrial membrane are responsible for ischemia-induced loss in activity. Palmitoyl-carnitine supported ADP-dependent state 3 respiration declined as a result of ischemia. When octanoyl-carnitine was utilized state 3 respiration remained unchanged. State 4 respiration increased during ischemia, an increase that appears specific to fatty utilization. Thus, VLCAD represents a likely site for the modulation of substrate utilization during myocardial ischemia. However, the dramatic increase in mitochondrial state 4 respiration would be predicted to accentuate the imbalance between energy production and utilization.

Keyword: glycolysis

Macrophages with a deletion of the () gene have a more proinflammatory phenotype.

Phosphoenolpyruvate carboxykinase (Pck1) is a metabolic enzyme that is integral to the gluconeogenic and glyceroneogenic pathways. However, Pck1\'s role in macrophage metabolism and function is unknown. Using stable isotopomer MS analysis in a mouse model with a myeloid cell-specific deletion, we show here that this deletion increases the proinflammatory phenotype in macrophages. Incubation of LPS-stimulated bone marrow-derived macrophages (BMDM) with [U-C]glucose revealed reduced C labeling of citrate and malate and increased C labeling of lactate in Pck1-deleted bone marrow-derived macrophages. We also found that the Pck1 deletion in the myeloid cells increases reactive oxygen species (ROS). Of note, this altered macrophage metabolism increased expression of the M1 cytokines TNFα, IL-1β, and IL-6. We therefore conclude that contributes to M1 polarization in macrophages. Our findings provide important insights into the factors determining the macrophage inflammatory response and indicate that Pck1 activity contributes to metabolic reprogramming and polarization in macrophages.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: glycolysis

Metformin influences cardiomyocyte cell death by pathways that are dependent and independent of caspase-3.

Metformin has been shown to increase fatty oxidation, an effect mediated by AMP activated protein kinase (AMPK). We hypothesised that metformin could prevent both caspase-3 activation and apoptosis when induced by .Cardiomyocytes were incubated with 1 mmol/l , in the absence or presence of metformin (1-5 mmol/l). Following 1 to 16 h, cell damage was evaluated by measuring lactate dehydrogenase released into the incubation medium, and Hoechst staining. To investigate the mechanism of metformin\'s effect on cardiomyocytes, substrate utilisation and phosphorylation of AMPK and acetyl-CoA carboxylase were measured. Intracellular mediators of apoptosis were also evaluated.Incubation of myocytes with for 16 h increased apoptosis, an effect that was partly blunted by 1 and 2 mmol/l metformin. This beneficial effect of metformin was associated with increased AMPK phosphorylation, oxidation and suppression of high-fat-induced increases in (1) long chain base biosynthesis protein 1 levels, (2) ceramide levels, and (3) caspase-3 activity. Unexpectedly, 5 mmol/l metformin dramatically increased apoptosis in myocytes incubated with high fat. This effect was associated with a robust increase in , lactate accumulation, and a significant drop of pH in the myocyte incubation medium.Our study demonstrates that metformin reduces high-fat-induced cardiac cell death, probably through inhibition of ceramide synthesis. However, at high concentrations, metformin causes proton and lactate accumulation, leading to cell damage that is independent of caspase-3.

Keyword: glycolysis

Altered metabolism causes cardiac dysfunction in perfused hearts from diabetic (db/db) mice.

Contractile function and substrate metabolism were characterized in perfused hearts from genetically diabetic C57BL/KsJ-lepr(db)/lepr(db) (db/db) mice and their non-diabetic lean littermates. Contractility was assessed in working hearts by measuring left ventricular pressures and cardiac power. Rates of , glucose oxidation, and fatty oxidation were measured using radiolabeled substrates ([5-(3)H]glucose, [U-(14)C]glucose, and [9,10-(3)H]palmitate) in the perfusate. Contractile dysfunction in db/db hearts was evident, with increased left ventricular end diastolic pressure and decreased left ventricular developed pressure, cardiac output, and cardiac power. The rate of from exogenous glucose in diabetic hearts was 48% of control, whereas glucose oxidation was depressed to only 16% of control. In contrast, palmitate oxidation was increased twofold in db/db hearts. The hypothesis that altered metabolism plays a causative role in diabetes-induced contractile dysfunction was tested using perfused hearts from transgenic db/db mice that overexpress GLUT-4 glucose transporters. Both glucose metabolism and palmitate metabolism were normalized in hearts from db/db-human insulin-regulatable glucose transporter (hGLUT-4) hearts, as was contractile function. These findings strongly support a causative role of impaired metabolism in the cardiomyopathy observed in db/db diabetic hearts.

Keyword: glycolysis

Recruitment of compensatory pathways to sustain oxidative flux with reduced carnitine palmitoyltransferase I activity characterizes inefficiency in energy metabolism in hypertrophied hearts.

Transport rates of long-chain free fatty acids into mitochondria via carnitine palmitoyltransferase I relative to overall oxidative rates in hypertrophied hearts remain poorly understood. Furthermore, the extent of glucose oxidation, despite increased in hypertrophy, remains controversial. The present study explores potential compensatory mechanisms to sustain tricarboxylic cycle flux that resolve the apparent discrepancy of reduced fatty oxidation without increased glucose oxidation through pyruvate dehydrogenase complex in the energy-poor, hypertrophied heart.We studied flux through the oxidative metabolism of intact adult rat hearts subjected to 10 weeks of pressure overload (hypertrophied; n=9) or sham operation (sham; n=8) using dynamic 13C-nuclear magnetic resonance. Isolated hearts were perfused with [2,4,6,8,10,12,14,16-(13)C8] palmitate (0.4 mmol/L) plus glucose (5 mmol/L) in a 14.1-T nuclear magnetic resonance magnet. At similar tricarboxylic cycle rates, flux through carnitine palmitoyltransferase I was 23% lower in hypertrophied (P<0.04) compared with sham hearts and corresponded to a shift toward increased expression of the L-carnitine palmitoyltransferase I isoform. Glucose oxidation via pyruvate dehydrogenase complex did not compensate for reduced palmitate oxidation rates. However, hypertrophied rats displayed an 83% increase in anaplerotic flux into the tricarboxylic cycle (P<0.03) that was supported by glycolytic pyruvate, coincident with increased mRNA transcript levels for malic enzyme.In cardiac hypertrophy, fatty oxidation rates are reduced, whereas compensatory increases in anaplerosis maintain tricarboxylic cycle flux and account for a greater portion of glucose oxidation than previously recognized. The shift away from acetyl coenzyme A production toward carbon influx via anaplerosis bypasses energy, yielding reactions contributing to a less energy-efficient heart.

Keyword: glycolysis

Role of glucose metabolism in the recovery of postischemic LV mechanical function: effects of insulin and other metabolic modulators.

The role of proton (H+) production from glucose metabolism in the recovery of myocardial function during postischemic reperfusion and its alteration by insulin and other metabolic modulators were examined. Rat hearts were perfused in vitro with Krebs-Henseleit solution containing palmitate (1.2 mmol/l) and glucose (11 mmol/l) under nonischemic conditions or during reperfusion following no-flow ischemia. Perfusate contained normal insulin (n-Ins, 50 mU/l), zero insulin (0-Ins), or supplemental insulin (s-Ins, 1,000 mU/l) or other metabolic modulators [dichloroacetate (DCA) at 3 mmol/l, oxfenicine at 1 mmol/l, and N6-cyclohexyladenosine (CHA) at 0.5 micromol/l]. Relative to n-Ins, 0-Ins depressed rates of and glucose oxidation in nonischemic hearts and impaired recovery of postischemic function. Relative to n-Ins, s-Ins did not affect aerobic glucose metabolism and did not improve recovery when present during reperfusion. When present during ischemia and reperfusion, s-Ins impaired recovery. Combinations of metabolic modulators with s-Ins stimulated glucose oxidation approximately 2.5-fold in nonischemic hearts and reduced H+ production. DCA and CHA, in combination with s-Ins, improved recovery of function, but addition of oxfenicine to this combination provided no further benefit. Although DCA and CHA were each partially protective in hearts perfused with n-Ins, optimal protection was achieved with DCA + CHA; recovery of function was inversely proportional to H+ production during reperfusion. Although supplemental insulin is not beneficial, elimination of H+ production from glucose metabolism by simultaneous inhibition of and stimulation of glucose oxidation optimizes recovery of postischemic mechanical function.

Keyword: glycolysis

Reversible high affinity inhibition of phosphofructokinase-1 by acyl-CoA: a mechanism integrating glycolytic flux with lipid metabolism.

The enzyme phosphofructokinase-1 (PFK-1) catalyzes the first committed step of and is regulated by a complex array of allosteric effectors that integrate glycolytic flux with cellular bioenergetics. Here, we demonstrate the direct, potent, and reversible inhibition of purified rabbit muscle PFK-1 by low micromolar concentrations of long chain fatty acyl-CoAs (apparent Ki∼1 μM). In sharp contrast, short chain acyl-CoAs, palmitoylcarnitine, and in the presence of CoASH were without effect. Remarkably, MgAMP and MgADP but not MgATP protected PFK-1 against inhibition by palmitoyl-CoA indicating that acyl-CoAs regulate PFK-1 activity in concert with cellular high energy phosphate status. Furthermore, incubation of PFK-1 with [1-(14)C]palmitoyl-CoA resulted in robust acylation of the enzyme that was reversible by incubation with acyl-protein thioesterase-1 (APT1). Importantly, APT1 reversed palmitoyl-CoA-mediated inhibition of PFK-1 activity. Mass spectrometric analyses of palmitoylated PFK-1 revealed four sites of acylation, including Cys-114, Cys-170, Cys-351, and Cys-577. PFK-1 in both skeletal muscle extracts and in purified form was inhibited by S-hexadecyl-CoA, a nonhydrolyzable palmitoyl-CoA analog, demonstrating that covalent acylation of PFK-1 was not required for inhibition. Tryptic footprinting suggested that S-hexadecyl-CoA induced a conformational change in PFK-1. Both palmitoyl-CoA and S-hexadecyl-CoA increased the association of PFK-1 with Ca2+/calmodulin, which attenuated the binding of palmitoylated PFK-1 to membrane vesicles. Collectively, these results demonstrate that fatty acyl-CoA modulates phosphofructokinase activity through both covalent and noncovalent interactions to regulate glycolytic flux and enzyme membrane localization via the branch point metabolic node that mediates lipid flux through anabolic and catabolic pathways.

Keyword: glycolysis

inhibition by palmitate in renal cells cultured in a two-chamber system.

A major shortcoming of renal proximal tubular cells (RPTC) in culture is the gradual modification of their energy metabolism from the oxidative type to the glycolytic type. To test the possible reduction of by naturally occurring long-chain fatty acids, RPTC were cultured in a two-chamber system, with albumin-bound palmitate (0.4 mM) added to the basolateral chamber after confluency. Twenty-four hours of contact with palmitate decreased by 38% provided that carnitine was present; lactate production was decreased by 38%, and the decrease in resulted from a similar decrease of basolateral and apical net uptake of glucose. In contrast to the previously described effect of the nonphysiological oxidative substrate heptanoate, palmitate promoted a long-term decrease in lactate production and sustained excellent cellular growth. After 4 days of contact, decreased was maintained even in the absence of carnitine and resulted from a decrease of basolateral uptake only, suggestive of long-term regulation different from the earlier effects. Thus, although cultured RPTC lost their oxidative phenotype, they exhibited a type of regulation (Randle effect) that is found in the oxidative-type but not in the glycolytic-type tissues, therefore unmasking a regulative capacity barely detectable in fresh RPTC. Low PO2 (50 mmHg in the apical chamber) could be a major cause of elevated and could hinder the effects of palmitate.

Keyword: glycolysis

Palmitate inhibits liver . Involvement of fructose 2,6-bisphosphate in the glucose/fatty cycle.

In hepatocytes from overnight-fasted rats incubated with glucose, palmitate decreased the production of lactate, the detritiation of [2-3H]- and [3-3H]-glucose, and the concentration of fructose 2,6-bisphosphate. Similarly, perfusion of hearts from fed rats with beta-hydroxybutyrate resulted in an inhibition of the detritiation of [3-3H]glucose and a fall in fructose 2,6-bisphosphate concentration. This fall could result from an increase in citrate (hepatocytes and heart) and sn-glycerol 3-bisphosphate concentration. It is suggested that a fall in fructose 2,6-bisphosphate concentration participates in the inhibition of by fatty acids and ketone bodies.

Keyword: glycolysis

Calcium regulation of , glucose oxidation, and fatty oxidation in the aerobic and ischemic heart.

Although Ca2+ is an important regulator of energy metabolism, the effects of increasing extracellular [Ca2+] on energy substrate preference are not clear. We determined the relationship between [Ca2+], fatty acids, and ischemia on rates of , glucose oxidation, and palmitate oxidation in isolated working rat hearts. Hearts were perfused with Krebs-Henseleit buffer containing 11 mM glucose, 100 microU/mL insulin, and either 1.25 or 2.5 mM Ca2+, in the presence or absence of 1.2 mM palmitate. Rates of and glucose oxidation or palmitate oxidation were measured in the hearts using [5-3H,14C(U)]glucose or [1-14C]palmitate, respectively. In the absence of fatty acids, and glucose oxidation rates were similar, regardless of whether [Ca2+ was 1.25 or 2.5 mM. Addition of 1.2 mM palmitate to the perfusate of hearts perfused with 1.25 mM Ca2+ significantly decreased rates of both (from 4623 +/- 438 to 1378 +/- 238 nmol.min-1.g-1 dry weight) and glucose oxidation (from 1392 +/- 219 to 114 +/- 22 nmol.min-1.g-1 dry weight). When [Ca2+] was increased from 1.25 to 2.5 mM in hearts perfused with 1.2 mM palmitate, and glucose oxidation increased by 164 and 271%, respectively, with no change in palmitate oxidation rates. Increasing [Ca2+] from 1.25 to 2.5 mM increased the contribution of glucose to ATP production from 9.3 to 18.7%. When hearts were subjected to low-flow ischemia (by reducing coronary flow to 0.5 mL.min-1) oxidative metabolism was essentially abolished. Under these conditions, glycolytic rates were not dependent on either [Ca2+] or the presence or absence of fatty acids. These results demonstrate that perfusate [Ca2+] is an important determinant of myocardial glucose metabolism in aerobic hearts, and that and glucose oxidation are more responsive to changes in [Ca2+] than is fatty oxidation.

Keyword: glycolysis

Effects of dichloroacetate on the metabolism of glucose, pyruvate, acetate, 3-hydroxybutyrate and palmitate in rat diaphragm and heart muscle in vitro and on extraction of glucose, lactate, pyruvate and free fatty acids by dog heart in vivo.

1. The extractions of glucose, lactate, pyruvate and free fatty acids by dog heart in vivo were calculated from measurements of their arterial and coronary sinus blood concentration. Elevation of plasma free fatty concentrations by infusion of intralipid and heparin resulted in increased extraction of free fatty acids and diminished extractions of glucose, lactate and pyruvate by the heart. It is suggested that metabolism of free fatty acids by the heart in vivo, as in vitro, may impair utilization of these substrates. These effects of elevated plasma free fatty concentrations on extractions by the heart in vivo were reversed by injection of dichloroacetate, which also improved extraction of lactate and pyruvate by the heart in vivo in alloxan diabetes. 2. Sodium dichloroacetate increased glucose oxidation and pyruvate oxidation in hearts from fed normal or alloxan-diabetic rats perfused with glucose and insulin. Dichloroacetate inhibited oxidation of acetate and 3-hydroxybutyrate and partially reversed inhibitory effects of these substrates on the oxidation of glucose. In rat diaphragm muscle dichloroacetate inhibited oxidation of acetate, 3-hydroxybutyrate and palmitate and increased glucose oxidation and pyruvate oxidation in diaphragms from alloxan-diabetic rats. Dichloroacetate increased the rate of in hearts perfused with glucose, insulin and acetate and evidence is given that this results from a lowering of the citrate concentration within the cell, with a consequent activation of phosphofructokinase. 3. In hearts from normal rats perfused with glucose and insulin, dichloroacetate increased cell concentrations of acetyl-CoA, acetylcarnitine and glutamate and lowered those of aspartate and malate. In perfusions with glucose, insulin and acetate, dichloroacetate lowered the cell citrate concentration without lowering the acetyl-CoA or acetylcarnitine concentrations. Measurements of specific radioactivities of acetyl-CoA, acetylcarnitine and citrate in perfusions with [1-(14)C]acetate indicated that dichloroacetate lowered the specific radio-activity of these substrates in the perfused heart. Evidence is given that dichloroacetate may not be metabolized by the heart to dichloroacetyl-CoA or dichloroacetylcarnitine or citrate or CO(2). 4. We suggest that dichloroacetate may activate pyruvate dehydrogenase, thus increasing the oxidation of pyruvate to acetyl-CoA and acetylcarnitine and the conversion of acetyl-CoA into glutamate, with consumption of aspartate and malate. Possible mechanisms for the changes in cell citrate concentration and for inhibitory effects of dichloroacetate on the oxidation of acetate, 3-hydroxybutyrate and palmitate are discussed.

Keyword: glycolysis

Acetyl-CoA from inflammation-induced fatty acids oxidation promotes hepatic malate-aspartate shuttle activity and .

Hepatic metabolic syndrome is associated with inflammation, as inflammation stimulates the reprogramming of nutrient metabolism and hepatic mitochondria-generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and fatty acids, in the condition of inflammation is still unclear. Here, we used an acute inflammation model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic and fatty oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs liver showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain . With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived fatty oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by fatty oxidation promotes MAS activity and via nonenzymatic acetylation during the inflammatory stress response.

Keyword: glycolysis

Effects of exposure to a simulated altitude of 5500 m on energy metabolic pathways in rats.

We examined the effect of exposure to 5500 m on three closely related metabolic pathways: anaerobic , the pentose phosphate shunt (PPS), and fatty metabolism. Rats were exposed to simulated altitude of 5500 m for up to 3 months. The maximal rate of lactate production in tissue homogenates, tissue lactic dehydrogenase and blood lactate levels were measured to evaluate the capacity for anaerobic . The uptake of 14C-1-palmitate, oxidation of 14C-1-palmitate to 14CO2, incorporation of 14C-1-palmitate into tissue lipids, plasma and tissue free fatty acids (FFA) levels and total lipid contents were measured to assess the magnitude of lipid metabolism. Activities of glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phophogluconate dehydrogenase (6-PGD) in the PPS pathway were measured to assess the capacity to generate reducing power. Acute and chronic hypoxia did not affect most of the measurements of anaerobic , but depressed lactate production in liver and kidney. Chronic hypoxia enhanced all aspects of lipid metabolism in liver and enhanced the uptake and oxidation to CO2 of palmitate in skeletal muscle. Chronic hypoxia did not alter the activity of the G-6-PD in any tissue studied, but the activity of 6-PGD was depressed in heart, kidney, thymus and adrenal gland. The lack of major changes in the capacities of anaerobic glycolytic pathways and the activities of the PPS dehydrogenases is consistent with the maintenance of normal aerobic metabolism in rats at 5500 m. We found no evidence that anaerobic metabolic processes were upregulated to sustain energy consumption during chronic hypoxia. On the other hand, enhanced fatty metabolism may spare carbohydrate for metabolic fuel under conditions of extreme hypoxic limitation.

Keyword: glycolysis

Tissue metabolomics of hepatocellular carcinoma: tumor energy metabolism and the role of transcriptomic classification.

Hepatocellular carcinoma (HCC) is one of the commonest causes of death from cancer. A plethora of metabolomic investigations of HCC have yielded molecules in biofluids that are both up- and down-regulated but no real consensus has emerged regarding exploitable biomarkers for early detection of HCC. We report here a different approach, a combined transcriptomics and metabolomics study of energy metabolism in HCC. A panel of 31 pairs of HCC tumors and corresponding nontumor liver tissues from the same patients was investigated by gas chromatography-mass spectrometry (GCMS)-based metabolomics. HCC was characterized by ∼2-fold depletion of glucose, glycerol 3- and 2-phosphate, malate, alanine, myo-inositol, and linoleic . Data are consistent with a metabolic remodeling involving a 4-fold increase in over mitochondrial oxidative phosphorylation. A second panel of 59 HCC that had been typed by transcriptomics and classified in G1 to G6 subgroups was also subjected to GCMS tissue metabolomics. No differences in glucose, lactate, alanine, glycerol 3-phosphate, malate, myo-inositol, or stearic tissue concentrations were found, suggesting that the Wnt/β-catenin pathway activated by CTNNB1 mutation in subgroups G5 and G6 did not exhibit specific metabolic remodeling. However, subgroup G1 had markedly reduced tissue concentrations of 1-stearoylglycerol, 1-palmitoylglycerol, and , suggesting that the high serum α-fetoprotein phenotype of G1, associated with the known overexpression of lipid catabolic enzymes, could be detected through metabolomics as increased lipid catabolism.Tissue metabolomics yielded precise biochemical information regarding HCC tumor metabolic remodeling from mitochondrial oxidation to aerobic and the impact of molecular subtypes on this process.Copyright © 2013 American Association for the Study of Liver Diseases.

Keyword: glycolysis

Cardiac efficiency is improved after ischemia by altering both the source and fate of protons.

Cardiac efficiency is decreased in hearts after severe ischemia. We determined whether reducing the production of H+ from glucose metabolism or inhibiting the clearance of H+ via Na(+)-H+ exchange could increase cardiac efficiency during reperfusion. This was achieved using dichloroacetate (DCA) to stimulate glucose oxidation and 5-(N,N-dimethyl)-amiloride (DMA) to inhibit Na(+)-H+ exchange, respectively. Isolated working rat hearts were subjected to 30 minutes of global ischemia and 60 minutes of reperfusion. and oxidation rates of glucose, lactate, and palmitate were measured. Recovery of cardiac work, O2 consumption (MVO2), and rates of acetyl-coenzyme A and ATP production during reperfusion were determined. After ischemia, cardiac work recovered to 35 +/- 5% of preischemic values in control hearts (n = 23), although MVO2, tricarboxylic (TCA) cycle activity, and ATP production from and oxidative metabolism rapidly recovered to preischemic levels. This decrease in cardiac efficiency was accompanied by a substantial production of H+ from glucose metabolism DCA caused a 2.2-fold increase in glucose oxidation, a 46 +/- 17% decrease in H+ production, a 1.6-fold increase in cardiac efficiency, and a 2.0-fold increase in cardiac work during reperfusion (n = 17). Inhibition of Na(+)-H+ exchange with DMA did not alter TCA cycle activity and ATP production rates but did result in a 1.8-fold increase in cardiac efficiency and a 1.7-fold increase in cardiac work (n = 12). These data show that cardiac efficiency and the contractile function after ischemia can be improved by either reducing the rate of H+ production from glucose metabolism during reperfusion or inhibiting the clearance of H+ via Na(+)-H+ exchange. Our data suggest that an increased requirement for ATP to restore ischemia-reperfusion-induced alterations in ion homeostasis contributes to the decrease in cardiac efficiency and contractile function after ischemia.

Keyword: glycolysis

Fatty oxidation by human platelets and its stimulation by thrombin.

Keyword: glycolysis

Investigating Cellular Quiescence of T Lymphocytes and Antigen-Induced Exit from Quiescence.

Naïve T cells are in a quiescent state under homeostasis but respond to antigen stimulation by exiting from quiescence and entering the cell cycle. Appropriate regulation of quiescence is crucial for maintaining T cell homeostasis at steady state and initiating proper T cell responses to antigen stimulation. Emerging evidence indicates that signaling by mechanistic target of rapamycin (mTOR) plays a central role in the control of T cell quiescence and antigen-induced exit from quiescence through coordinating immune signals, cellular metabolic programs, and cell cycle machinery. The mTOR-dependent regulation of quiescence has also been implicated in the differentiation and function of memory T cells. In this chapter, we describe techniques to assess quiescent state of naïve T cells under steady state and exit from quiescence upon TCR stimulation.

Keyword: glycolysis

Uncovering sperm metabolome to discover biomarkers for bull fertility.

Subfertility decreases the efficiency of the cattle industry because artificial insemination employs spermatozoa from a single bull to inseminate thousands of cows. Variation in bull fertility has been demonstrated even among those animals exhibiting normal sperm numbers, motility, and morphology. Despite advances in research, molecular and cellular mechanisms underlying the causes of low fertility in some bulls have not been fully elucidated. In this study, we investigated the metabolic profile of bull spermatozoa using non-targeted metabolomics. Statistical analysis and bioinformatic tools were employed to evaluate the metabolic profiles high and low fertility groups. Metabolic pathways associated with the sperm metabolome were also reported.A total of 22 distinct metabolites were detected in spermatozoa from bulls with high fertility (HF) or low fertility (LF) phenotype. The major metabolite classes of bovine sperm were organic acids/derivatives and fatty acids/conjugates. We demonstrated that the abundance ratios of five sperm metabolites were statistically different between HF and LF groups including gamma-aminobutyric (GABA), carbamate, benzoic , lactic , and . Metabolites with different abundances in HF and LF bulls had also VIP scores of greater than 1.5 and AUC- ROC curves of more than 80%. In addition, four metabolic pathways associated with differential metabolites namely alanine, aspartate and glutamate metabolism, β-alanine metabolism, or gluconeogenesis, and pyruvate metabolism were also explored.This is the first study aimed at ascertaining the metabolome of spermatozoa from bulls with different fertility phenotype using gas chromatography-mass spectrometry. We identified five metabolites in the two groups of sires and such molecules can be used, in the future, as key indicators of bull fertility.

Keyword: glycolysis

Skeletal muscle lipids and glycogen mask substrate competition (Randle cycle).

The glucose-free fatty (FFA) cycle (Randle) was examined in soleus muscle, a red muscle with a high lipid oxidation rate, and extensor digitorum longus (EDL) muscle, a white muscle with a low lipid oxidation rate, using a carnitine palmethyltransferase (CPT-I) inhibitor as a probe. Exogenous palmitate by itself had little if any effect on or glycogen accumulation in the two muscle types. The CPT-I inhibitor markedly decreased glycogen accumulation in both muscles (from fed rats), but increased (lactate formation) and glucose oxidation to carbon dioxide only in the red muscle. When the muscles were made more dependent on FFA oxidation by prior fasting or exercise, the CPT-I stimulatory effect on and glucose oxidation in white muscle was unmasked. In conclusion, the competition between lipid and carbohydrate utilization (Randle cycle) is easily demonstrated in both red and white muscle using a CPT-I inhibitor as a probe. The difficulties encountered in showing this competition in other studies using exogenous FFA may be explained by a combination of factors, including (1) low tissue lipid oxidation rates, (2) competition between exogenous and endogenous lipids such that provision of exogenous lipids fails to increase overall lipid oxidation, and (3) preferential utilization of exogenous glucose with glycogen sparing in the presence of FFA.

Keyword: glycolysis

Glycerolipid biosynthesis in isolated rat intestinal epithelial cells.

Intestinal epithelial cells were prepared from fasted rats by dispersion with collagenase (EC 3.4.24.3). The structural and metabolic integrity of the cells was verified by electron microscopy, a high percentage of Trypan Blue exclusion, a low degree of release of lactate dehydrogenase (EC 1.1.1.27) in the medium, and by the retention of sensitivity to agents known to modify metabolic and transport activity in everted sacs of intestinal mucosa. The isolated intestinal epithelial cells were used to study glycerolipid biosynthesis from glucose, glycerol, 2-monoacylglycerol, and free fatty acids. The cells actively incorporated the labeled precursors into glycerolipids without specific cofactor requirements. Addition of fatty acids stimulated the incorporation of both glucose and glycerol into triacylglycerols and glycerophospholipids, the greatest effect being observed with palmitate. The stimulation of monoacylglycerol acylation appeared to depend on both the nature of the monoacylglycerol and fatty supplied. Stereospecific analyses of the diacylglycerols formed from 2-monoacylglycerols and free fatty acids showed that 1,2-diacyl-sn-glycerols (62-70%) were the major and that 2,3-diacyl-sn-glycerols (30-38%) the minor intermediates in triacylglycerol biosynthesis. The data indicate that isolated intestinal epithelial cells exhibit a total capacity of glycerolipid synthesis and a stereochemical course of reaction which is comparable to that observed for triacylglycerol formation in everted sacs of intestinal mucosa, but much less specific than that seen in microsomal preparations of intestinal mucosa.

Keyword: glycolysis

Carvedilol suppresses fatty oxidation and stimulates in C2C12 cells.

Beta adrenergic receptor blocking drugs (β-blockers) are used chronically in many cardiovascular diseases such as hypertension, ischemic heart disease, arrhythmia, and heart failure. Beneficial effects are associated with the inhibition of symphathetic nervous system hyperactivity, reduction of heart rate, and remodeling by blocking the mitogenic activity of catecholamines. A possible effect of β-blockers on substrate metabolism has also been suggested. The direct effects of β-blockers on mouse C2C12 cells were investigated in this study. C2C12 cells were grown in Dulbecco\'s modified Eagle\'s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and differentiated into myotubes in the same medium that contained 1% FBS. oxidation and were measured by using [9,10-(3)H]palmitate and [5-(3)H]glucose, respectively. The amount of (3)H(2)O was measured as an indicator of substrate usage. Carvedilol (100 µmol/L) inhibited palmitate oxidation and increased by nearly 50%. Prazosin altered substrate metabolism in a similar fashion as carvedilol, whereas propranolol or bisoprolol were devoid of metabolic effects. When added to mimic sympathetic activation, epinephrine stimulated but did not alter fatty oxidation. Based on these results, carvedilol appears to have direct effects on substrate metabolism that are related to the blockade of α1 adrenergic receptors.

Keyword: glycolysis

[Carbohydrate and metabolism in the myocardium of the hypoxic rat].

Keyword: glycolysis

Effect of ATP depletion on the palmitoylation of myelin proteolipid protein in young and adult rats.

The present study was designed to determine whether the palmitoylation of the hydrophobic myelin proteolipid protein (PLP) is dependent on cellular energy. To this end, brain slices from 20- and 60-day-old rats were incubated with [3H]palmitate for 1 h in the presence or absence of various metabolic poisons. In adult rats, the inhibition of mitochondrial ATP production with KCN (5 mM), oligomycin (10 microM), or rotenone (10 microM) reduced the incorporation of [3H]palmitate into fatty acyl-CoA and glycerolipids by 50-60%, whereas the labeling of PLP was unaltered. Incubation in the presence of rotenone (10 microM) plus NaF (5 mM) abolished the synthesis of acyl-CoA and lipid palmitoylation, but the incorporation of [3H]palmitate into PLP was still not different from that in controls. In rapidly myelinating animals, the inhibition of both mitochondrial electron transport and obliterated the palmitoylation of lipids but reduced that of PLP by only 40%. PLP acylation was reduced to a similar extent when slices were incubated for up to 3 h, indicating that exogenously added palmitate is incorporated into PLP by ATP-dependent and ATP-independent mechanisms. Determination of the number of PLP molecules modified by each of these reactions during development suggests that the ATP-dependent process is important during the formation and/or compaction of the myelin sheath, whereas the ATP-independent mechanism is likely to play a role in myelin maintenance, perhaps by participating in the periodic repair of thioester linkages between the fatty acids and the protein.

Keyword: glycolysis

Glucose deprivation in tuberous sclerosis complex-related tumors.

Cancer cells possess unique metabolic phenotypes that are determined by their underlying oncogenic pathways. Activation of the PI3K/Akt/mTOR signaling cascade promotes and leads to glucose-dependence in tumors. In particular, cells with constitutive mTORC1 activity secondary to the loss of TSC1/TSC2 function are prone to undergo apoptosis upon glucose withdrawal in vitro, but this concept has not been tested in vivo. This study examines the effects of restricting glucose metabolism by pharmacologic and dietary means in a tuberous sclerosis complex (TSC) tumor xenograft model.Tumor-bearing mice were randomly assigned to receive unrestricted carbohydrate-free ("Carb-free") or Western-style diet in the absence or presence of 2-deoxyglucose (2-DG) in one of four treatment groups. After 14 weeks, tumor sizes were significantly different among the four treatment groups with those receiving 2-DG having the smallest tumors. Unexpectedly, the "Carb-free" diet was associated with the largest tumors but they remained responsive to 2-DG. PET imaging showed significant treatment-related changes in tumor 18fluorodeoxyglucose-uptake but the standard uptake values did not correlate with tumor size. Alternative energy substrates such as ketone bodies and monounsaturated oleic supported the growth of the Tsc2-/- cells in vitro, whereas saturated was toxic. Correspondingly, tumors in the high-fat, "Carb-free" group showed greater necrosis and liquefaction that contributed to their larger sizes. In contrast, 2-DG treatment significantly reduced tumor cell proliferation, increased metabolic stress (i.e., ketonemia) and AMPK activity, whereas rapamycin primarily reduced cell size.Our data support the concept of glycolytic inhibition as a therapeutic approach in TSC whereas dietary withdrawal of carbohydrates was not effective.

Keyword: glycolysis

Influence of vanadate on , intracellular sodium, and pH in perfused rat hearts.

Vanadium compounds have been shown to cause a variety of biological and metabolic effects including inhibition of certain enzymes, alteration of contractile function, and as an insulin like regulator of glucose metabolism. However, the influence of vanadium on metabolic and ionic changes in hearts remains to be understood. In this study we have examined the influence of vanadate on glucose metabolism and sodium transport in isolated perfused rat hearts. Hearts were perfused with 10 mM glucose and varying vanadate concentrations (0.7-100 microM) while changes in high energy phosphates (ATP and phosphocreatine (PCr)), intracellular pH, and intracellular sodium were monitored using 31P and 23Na NMR spectroscopy. Tissue lactate, glycogen, and (Na+, K+)-ATPase activity were also measured using biochemical assays. Under baseline conditions, vanadate increased tissue glycogen levels two fold and reduced (Na+, K+)-ATPase activity. Significant decreases in ATP and PCr were observed in the presence of vanadate, with little change in intracellular pH. These changes under baseline conditions were less severe when the hearts were perfused with glucose, palmitate and beta-hydroxybutyrate. During ischemia vanadate did not limit the rise in intracellular sodium, but slowed sodium recovery on reperfusion. The presence of vanadate during ischemia resulted in attenuation of acidosis, and reduced lactate accumulation. Reperfusion in the presence of vanadate resulted in a slower ATP recovery, while intracellular pH and PCr recovery was not affected. These results indicate that vanadate alters glucose utilization and (Na+, K+)-ATPase activity and thereby influences the response of the myocardium to an ischemic insult.

Keyword: glycolysis

The effect of AMP-activated protein kinase and its activator AICAR on the metabolism of human umbilical vein endothelial cells.

In several non-vascular tissues in which it has been studied, AMP-activated protein kinase (AMPK) appears to modulate the cellular response to stresses such as ischemia. In liver and muscle, it phosphorylates and inhibits acetyl CoA carboxylase (ACC), leading to an increase in fatty oxidation; and in muscle, its activation is associated with an increase in glucose transport. Here we report the presence of both AMPK and ACC in human umbilical vein endothelial cells (HUVEC). Incubation of HUVEC with 2 mM AICAR, an AMPK activator, caused a 5-fold activation of AMPK, which was accompanied by a 70% decrease in ACC activity and a 2-fold increase in fatty oxidation. Surprisingly, glucose uptake and , the dominant energy-producing pathway in HUVEC, were diminished by 40-60%. Despite this, cellular ATP levels were increased by 35%. Thus activation of AMPK by AICAR is associated with major alterations in endothelial cell energy balance. Whether these alterations protect the endothelium during ischemia or other stresses remains to be determined.Copyright 1999 Academic Press.

Keyword: glycolysis

Reversible and irreversible oxidant injury to PC12 cells by hydrogen peroxide.

A simple and reproducible model to identify biochemical changes associated with the transition from reversible to irreversible oxidant injury and cell death was established using rat pheochromocytoma PC12 cells. Cells were subjected to a transient oxidative stress induced by exposure to hydrogen peroxide (H2O2). Reversible loss of high-energy phosphates, induced by exposing cells to 0.2 mM H2O2, was preceded by transient increases in cytosolic calcium with no loss of plasma membrane integrity, as indexed by release of cytosolic enzymes. In contrast, permanent loss of high-energy phosphates, induced by treating cells with 0.5 mH H2O2, was associated with sustained rises in cytosolic-free calcium and increased oxidation of pyruvate and palmitate, two mitochondrial substrates. Initial production of pyruvate and lactate was inhibited by exposure to 0.5 mM H2O2 but returned to values comparable to control values at one hour after treatment with H2O2. Compromise of the plasma membrane was a late event, occurring between 1 and 2 hours after exposure to 0.5 mM H2O2. Collectively, these data indicate that irreversible loss of high-energy phosphates and cell death caused by oxidative stress is more closely associated with altered mitochondrial function than with impaired .

Keyword: glycolysis

Reductive glutamine metabolism by IDH1 mediates lipogenesis under hypoxia.

Acetyl coenzyme A (AcCoA) is the central biosynthetic precursor for fatty- synthesis and protein acetylation. In the conventional view of mammalian cell metabolism, AcCoA is primarily generated from glucose-derived pyruvate through the citrate shuttle and ATP citrate lyase in the cytosol. However, proliferating cells that exhibit aerobic and those exposed to hypoxia convert glucose to lactate at near-stoichiometric levels, directing glucose carbon away from the tricarboxylic cycle and fatty- synthesis. Although glutamine is consumed at levels exceeding that required for nitrogen biosynthesis, the regulation and use of glutamine metabolism in hypoxic cells is not well understood. Here we show that human cells use reductive metabolism of α-ketoglutarate to synthesize AcCoA for lipid synthesis. This isocitrate dehydrogenase-1 (IDH1)-dependent pathway is active in most cell lines under normal culture conditions, but cells grown under hypoxia rely almost exclusively on the reductive carboxylation of glutamine-derived α-ketoglutarate for de novo lipogenesis. Furthermore, renal cell lines deficient in the von Hippel-Lindau tumour suppressor protein preferentially use reductive glutamine metabolism for lipid biosynthesis even at normal oxygen levels. These results identify a critical role for oxygen in regulating carbon use to produce AcCoA and support lipid synthesis in mammalian cells.

Keyword: glycolysis

Cardiac phosphatase-deficient 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase increases , hypertrophy, and myocyte resistance to hypoxia.

During ischemia and heart failure, there is an increase in cardiac . To understand if this is beneficial or detrimental to the heart, we chronically elevated by cardiac-specific overexpression of phosphatase-deficient 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2) in transgenic mice. PFK-2 controls the level of fructose-2,6-bisphosphate (Fru-2,6-P2), an important regulator of phosphofructokinase and . Transgenic mice had over a threefold elevation in levels of Fru-2,6-P2. Cardiac metabolites upstream of phosphofructokinase were significantly reduced, as would be expected by the activation of phosphofructokinase. In perfused hearts, the transgene caused a significant increase in that was less sensitive to inhibition by palmitate. Conversely, oxidation of palmitate was reduced by close to 50%. The elevation in made isolated cardiomyocytes highly resistant to contractile inhibition by hypoxia, but in vivo the transgene had no effect on ischemia-reperfusion injury. Transgenic hearts exhibited pathology: the heart weight-to-body weight ratio was increased 17%, cardiomyocyte length was greater, and cardiac fibrosis was increased. However, the transgene did not change insulin sensitivity. These results show that the elevation in provides acute benefits against hypoxia, but the chronic increase in or reduction in fatty oxidation interferes with normal cardiac metabolism, which may be detrimental to the heart.

Keyword: glycolysis

Regulation of ether lipids and their precursors in relation to in cultured neoplastic cells.

Tumors typically show high rates of and elevated levels of ether lipids, particularly the alkyldiacylglycerols; thus, we investigated the relationship between ether lipid accumulation and glucose metabolism in a neoplastic cell line (B2-1). The B2-1 cells grown in 5.5 mM galactose in the absence of glucose produced very low levels of alkyldiacylglycerols, triacylglycerols, lactic , and dihydroxyacetone-P. Increasing concentrations of glucose caused a progressive increase in lactic , dihydroxyacetone-P, and up to a ten-fold increase in alkyldiacylglycerols and triacylglycerols. Glucose supplements also caused an increased incorporation of [9,10-3H] into alkyldiacylglycerols and triacylglycerols. These metabolic changes appeared to be independent of altered growth rates of the cells. The addition of hexadecanol along with glucose to the cultures resulted in a shorter lag and a more rapid rate of accumulation of alkyldiacylglycerols; hexadecanol supplements alone had no effect. The extent of uptake and oxidation of hexadecanol was similar in both the glucose and galactose-grown cells. These results indicate that the levels of alkyldiacylglycerols in neoplastic cells can be regulated by the extent their precursors are formed from glucose.

Keyword: glycolysis

is predominant source of myocardial ATP production immediately after birth.

Glycolytic flux, as well as glucose, fatty , and lactate oxidation, was determined in isolated working hearts obtained from 1- and 7-day-old rabbits. One-day-old rabbit hearts were perfused via the inferior cava against a constant aortic and pulmonary arterial afterload, whereas hearts from 7-day-old rabbits were perfused via the left atria against a constant aortic afterload. Hearts were perfused with buffer containing 100 microU/ml insulin and either 1) 11 mM [U-14C/2-3H]glucose, 0.4 mM palmitate, 2 mM lactate; 2) 11 mM glucose, 0.4 mM [1-14C]palmitate, 2 mM lactate; or 3) 11 mM glucose, 0.4 mM palmitate, 2 mM [U-14C]lactate. Glycolytic rates (measured as 3H2O production) were high in 1-day-old hearts but decreased by 7 days (from 2,730 +/- 280 to 580 +/- 80 nmol.min-1.g dry wt-1). Rates of glucose oxidation (measured as 14CO2 production) were lower in both 1- and 7-day-old hearts (59 +/- 4.4 and 23 +/- 2 nmol.min-1.g dry wt-1). Palmitate oxidation rates were low in 1-day-old hearts but dramatically increased by 7 days (22.6 +/- 5.6 and 305 +/- 33 nmol oxidized.min-1.g dry wt-1, respectively). In contrast, lactate was readily oxidized by both 1- and 7-day-old hearts (169 +/- 14 and 456 +/- 52 nmol.min-1.g dry wt-1, respectively). In 1-day-old hearts, 44% of steady-state ATP production from exogenous sources were derived from , whereas 18, 13, and 25% were derived from glucose, palmitate, and lactate oxidation, respectively.(ABSTRACT TRUNCATED AT 250 WORDS).

Keyword: glycolysis

Demand-induced ischemia in volume expanded isolated rat heart; the effect of dichloroacetate and trimetazidine.

In failing hearts, coronary flow is normal, but the coronary flow reserve (CFR) is reduced, so demand-induced ischemia (DII) may occur in response to greater demand for O(2). The objectives of this study were: (i) to verify that dobutamine stimulation produces DII in isolated rat hearts having, like failing hearts, increased left ventricular end-diastolic pressure (LVEDP) and hence reduced CFR and (ii) to study the effects of stimulation of glucose oxidation and of inhibition of fatty oxidation in this new model of DII. Isolated rat hearts perfused with 11 mM glucose and 0.6 mM palmitate (or no palmitate) were studied. Stepwise increments in the volume of a balloon placed in LV resulted in reciprocal impairment of CFR, supporting the role of the extravascular compressive forces in determining CFR. CFR was 1.82+/-0.1 and 1.32+/-0.1 (p<0.05) in the hearts with LVEDP set to 5 mmHg (controls) and 40 mmHg (expanded), respectively. In controls, dobutamine increased coronary flow, myocardial oxygen consumption (MVO(2)), LVDP, mechanical efficiency, and the rates of palmitate and glucose oxidation, however, the effluent lactate concentration remained unchanged. In the expanded hearts vs. controls, dobutamine-induced increases in coronary flow and MVO(2) were reduced by approximately 50%, the increases in LVDP, efficiency, and rates of glucose and fatty oxidation were completely prevented, and lactate production greatly increased with dobutamine, indicating DII. Pyruvate dehydrogenase activator, dichloroacetate (DCA 1 mM) and a putative inhibitor of fatty beta-oxidation, trimetazidine (5 microM), both increased the rate of glucose oxidation and attenuated myocardial lactate production during DII, however they did not improve myocardial function during DII. Likewise, palmitate-free perfusion had no beneficial effect during DII although it attenuated lactate production. In the hearts subjected to palmitate-free perfusion plus DCA, lactate overproduction during DII was completely abolished, however, the deterioration of LVDP and mechanical efficiency was only partially prevented. Thus, greater demand for O(2) induces DII in the expanded hearts with reduced CFR. Lactate overproduction secondary to an imbalance between and glucose oxidation is not a primary factor adversely affecting cardiac mechanical function during DII. Interventions shifting this balance toward glucose oxidation are not beneficial in the setting of DII in our model although they are known to effectively mitigate contractile dysfunction in the post-ischemic myocardium.

Keyword: glycolysis

Metabolomics changes in a rat model of obstructive jaundice: mapping to metabolism of amino acids, carbohydrates and lipids as well as oxidative stress.

The study examined the global metabolic and some biochemical changes in rats with cholestasis induced by bile duct ligation (BDL). Serum samples were collected in male Wistar rats with BDL (n\xa0=\xa08) and sham surgery (n\xa0=\xa08) at day 3 after surgery for metabolomics analysis using a combination of reversed phase chromatography and hydrophilic interaction chromatography (HILIC) and quadrupole-time-of-flight mass spectrometry (Q-TOF MS). The serum levels of malondialdehyde (MDA), total antioxidative capacity (T-AOC), glutathione (GSH) and glutathione disulfide (GSSG), the activities of superoxide dismutase (SOD) and glutathion peroxidase (GSH-Px) were measured to estimate the oxidative stress state. Key changes after BDL included increased levels of l-phenylalanine, l-glutamate, l-tyrosine, kynurenine, l-lactic , LysoPC(c) (14:0), glycine and succinic and decreased levels of l-valine, PC(b) (19:0/0:0), taurine, , l-isoleucine and citric metabolism products. And treatment with BDL significantly decreased the levels of GSH, T-AOC as well as SOD, GSH-Px activities, and upregulated MDA levels. The changes could be mapped to metabolism of amino acids and lipids, Krebs cycle and , as well as increased oxidative stress and decreased antioxidant capability. Our study indicated that BDL induces major changes in the metabolism of all 3 major energy substances, as well as oxidative stress.

Keyword: glycolysis

A hypothetical model to solve the controversy over the involvement of UCP2 in palmitate-induced β-cell dysfunction.

The aim of this article is to solve an existing controversy over the involvement of uncoupling protein-2 in the impairment of glucose-stimulated insulin secretion induced by chronic exposure of β-cells to palmitate. We analyzed and compared the results of studies that support and that deny the involvement of uncoupling protein-2 in this impairment. We observed that this impairment could occur in multiple stages. We provide a model in which palmitate-induced impairment of glucose-stimulated insulin secretion is proposed to occur in two stages, early stage and late stage, depending on the integrity of electron supply ( and Krebs cycle) and transport system through electron transport chain after palmitate treatment. Prolonged exposure of β-cells to palmitate can impair this system. Early-stage impairment occurs due to uncoupling by uncoupling protein-2 when this system is still intact. When this system becomes impaired, late-stage impairment occurs mainly due to reduced glucose-stimulated adenosine triphosphate production independent of uncoupling by uncoupling protein-2. The change in glucose-stimulated oxygen uptake after palmitate treatment reflects the integrity of this system and can be used to differentiate between the two stages. Some β-cells lines and islets appear to be more resistant to palmitate-induced impairment of electron supply and transport system than others, and therefore early stage is prominent in the more resistant cell lines and less prominent or absent in the less resistant cell lines. This may help to resolve the pathogenesis of diabetes and to monitor the progression of palmitate-induced β-cell dysfunction.

Keyword: glycolysis

Metabolism of freshly isolated human hair follicles capable of hair elongation: a glutaminolytic, aerobic glycolytic tissue.

The metabolism of the human hair follicle was investigated in vitro under conditions that maintained glycogen and adenosine triphosphate (ATP) content and the growth rate of the follicle at values observed in vivo. We have shown that only 10% of the total glucose utilized was oxidized to CO2 and 40% of this was oxidized via the pentose phosphate shunt. Although fatty acids and ketone bodies were oxidized by the hair follicle, they are poor energetic substitutes for glucose. Nor will fatty acids or ketone bodies sustain hair growth in vitro. Glutamine, however, was shown, both biochemically and by comparing growth rates, to be an important fuel with 23% of uptake being oxidized, generating a possible 2.16 +/- 0.32 nmoles ATP/follicle/h (mean +/- SEM) (glucose metabolism generates 4.54 +/- 0.61 nmoles ATP/follicle/h). Sixty-four percent of the glutamine taken up was calculated to be metabolized to lactate, showing that the hair follicle engages in both and glutaminolysis. The glucose-fatty cycle appears to be unimportant in the hair follicle but our data indicates that a glucose-glutamine cycle does operate.

Keyword: glycolysis

Acute effects of triiodothyronine on glucose and fatty metabolism during reperfusion of ischemic rat hearts.

Clinical studies have demonstrated improved myocardial recovery after severe ischemia in response to acute triiodothyronine (T3) treatment. We determined whether T3 improves the recovery of ischemic hearts by improving energy substrate metabolism. Isolated working rat hearts were perfused with 5.5 mM glucose and 1.2 mM palmitate and were subjected to 30 min of no-flow ischemia. , glucose oxidation, and palmitate oxidation were measured during aerobic reperfusion by adding [5-3H]glucose, [U-14C]glucose, or [9,10-3H]palmitate to the perfusate, respectively. During reperfusion, cardiac work in untreated hearts recovered to a lesser extent than myocardial O2 consumption (MVO2), resulting in a decreased recovery of cardiac efficiency, which recovered to only 25% of preischemic values. Treatment of hearts with T3 (10 nM) before ischemia increased glucose oxidation during reperfusion, which was associated with a significant increase in pyruvate dehydrogenase (PDH) activity, the rate-limiting enzyme for glucose oxidation. In contrast, T3 had no effect on MVO2, , or palmitate oxidation. This resulted in a significant decrease in H+ production from uncoupled from glucose oxidation (2.7 +/- 0.3 and 1.9 +/- 0.3 micromol . g dry wt-1 . min-1 in control and T3-treated hearts, respectively, P < 0.05), as well as a 3.2-fold improvement in cardiac work and a 2.3-fold increase in cardiac efficiency compared with untreated postischemic hearts (P < 0.05). These data suggest that T3 can exert acute effects that improve the coupling of to glucose oxidation, thereby decreasing H+ production and increasing cardiac efficiency as well as contractile function during reperfusion of the postischemic heart.

Keyword: glycolysis

Beta-cell hypersensitivity to glucose following 24-h exposure of rat islets to fatty acids.

Prolonged exposure of islets to fatty acids results in a lowered glucose set-point for insulin secretion. We examined the mechanism in islets cultured for 24 h with 0.25 mmol/l palmitate. As expected, insulin secretion at 2.8 and 8.3 mmol/l glucose was increased in the palmitate-treated islets as opposed to no change at 27.7 mmol/l glucose. Co-culturing with 0.05 microgram/ml Triacsin C, an inhibitor of long chain acyl-CoA synthetase, blocked this effect. Glucose utilization and oxidation showed the same pattern as insulin secretion, with the step-up for both measurements being fully manifest at 2.8 mmol/l glucose. Glucokinase Km and Vmax measured in islet extracts were unaffected by the palmitate. In contrast, hexokinase Vmax was increased by 25-35% in both the cytoplasmic and mitochondrial-bound pools. Our data suggest prolonged exposure to fatty acids increased beta-cell hexokinase activity, thereby modifying the kinetics of glucose entry into the metabolic pathway and glucose-induced insulin secretion. The cellular mediator is likely an increased level of long chain fatty acyl-CoA esters.

Keyword: glycolysis

Gene coexpression network analysis of oil biosynthesis in an interspecific backcross of oil palm.

Global demand for vegetable oils is increasing at a dramatic rate, while our understanding of the regulation of oil biosynthesis in plants remains limited. To gain insights into the mechanisms that govern oil synthesis and fatty (FA) composition in the oil palm fruit, we used a multilevel approach combining gene coexpression analysis, quantification of allele-specific expression and joint multivariate analysis of transcriptomic and lipid data, in an interspecific backcross population between the African oil palm, Elaeis guineensis, and the American oil palm, Elaeis oleifera, which display contrasting oil contents and FA compositions. The gene coexpression network produced revealed tight transcriptional coordination of fatty synthesis (FAS) in the plastid with sugar sensing, plastidial glycolysis, transient starch storage and carbon recapture pathways. It also revealed a concerted regulation, along with FAS, of both the transfer of nascent FA to the endoplasmic reticulum, where triacylglycerol assembly occurs, and of the production of glycerol-3-phosphate, which provides the backbone of triacylglycerols. Plastid biogenesis and auxin transport were the two other biological processes most tightly connected to FAS in the network. In addition to WRINKLED1, a transcription factor (TF) known to activate FAS genes, two novel TFs, termed NF-YB-1 and ZFP-1, were found at the core of the FAS module. The saturated FA content of palm oil appeared to vary above all in relation to the level of transcripts of the gene coding for β-ketoacyl-acyl carrier protein synthase II. Our findings should facilitate the development of breeding and engineering strategies in this and other oil crops.© 2016 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

Keyword: glycolysis

Performance of the neonatal pig heart subjected to oxygen insufficiency.

Isolated, paced, isovolumically beating, neonatal pig (n = 32) hearts underwent retrograde aortic perfusion with a solution containing insulin (100 microU/ml), glucose (5.5 mM), and palmitate (0.55 mM). , lactate release, glucose oxidation, palmitate oxidation, and oxygen consumption were assessed. The hearts were perfused during three periods: (1) baseline, pO(2) approximately 500 mm Hg, heart rate 150 bpm; (2) hypoxia, pO(2) approximately 60-80 mm Hg, heart rate 150 bpm, or tachycardia, pO(2) approximately 500 mm Hg, heart rate 300 bpm, and (3) recovery, return to baseline conditions. For hypoxia and tachycardia, the oxygen supply-demand ratio was comparable ( approximately 1 nmol O(2)/mm Hg/g(dry)). During baseline, the left ventricular peak systolic pressure (PSP) averaged 126 +/- 6 mm Hg, the end-diastolic pressure (EDP) 5 mm Hg, and the relaxation time constant (Tau) 34 +/- 2 ms; the coronary flow was 36 +/- 2 ml/min/g(dry). During hypoxia, the PSP decreased to 70 +/- 2 mm Hg, while EDP, Tau, and coronary flow increased to 26 +/- 2 mm Hg, 104 +/- 14 ms, and 70 +/- 2 ml/min/g(dry), respectively; palmitate oxidation and oxygen consumption decreased well below baseline. During tachycardia, the PSP decreased to 88 +/- 1 mm Hg, and the EDP increased to 11 +/- 1 mm Hg, while Tau and coronary flow did not change significantly; palmitate oxidation and oxygen consumption increased above baseline. For both stressors, the predicted lactate release underestimated the measured values by a factor of approximately 2, but were comparable during baseline and recovery. Upon recovery, PSP returned to approximately 80% of baseline, while EDP remained elevated, for both stressors. Glucose oxidation returned to baseline, but palmitate oxidation became accelerated. We conclude for neonatal pig hearts subjected to oxygen insufficiency: (1) that PSP decreases and (2) that EDP and Tau increase with hypoxia, whereas EDP increases, while Tau remains unchanged with tachycardia. Following both stressors, palmitate oxidation becomes enhanced and dissociated from mechanical activity.Copyright 2004 S. Karger AG, Basel

Keyword: glycolysis

Fatty composition of individual plasma steryl esters in phytosterolemia and xanthomatosis.

The bulk of the plasma plant sterol in phytosterolemia occurs in the esterified form and is carried mostly in the low and high density lipoproteins. We have determined the fatty composition of the individual plasma steryl esters from a newly discovered subject with phytosterolemia and xanthomatosis. For this purpose the intact steryl esters were subject to high temperature gas liquid chromatography (GLC) on a polar capillary column, which separated the major esters on the basis of molecular weight and degree of unsaturation of the fatty acids. The saturated and unsaturated sterols esterified to saturated, monoenic, dienoic and tetraenoic fatty acids were identified by GLC analysis of the sterol moieties of the corresponding AgNO3-TLC fractions of the steryl esters. The GLC results were confirmed by reversed phase high performance liquid chromatography combined with mass spectrometry via direct liquid inlet interface. It was found that, in general, each fatty was esterified to the same complement of sterols, and that the esterified sterols possessed a composition comparable to that of the free plasma sterols, which was comprised of about 75% cholesterol, 6% campesterol, 4% 22,23-dihydrobrassicasterol and 15% beta-sitosterol. The fatty composition of the steryl esters differed from that of the 2-position of the plasma phosphatidylcholines, which contained significantly less and oleic and more linoleic . On the basis of these results and a review of the literature it is suggested that the plasma cholesteryl and plant steryl esters in phytosterolemia originate from both synthesis in plasma via the lecithin-cholesterol acyltransferase and synthesis in tissues via the acylCoA-cholesterol acyltransferase.

Keyword: hyperlipedemia

EGFR Inhibition Blocks -induced inflammation in cardiomyocytes and Prevents -induced Cardiac Injury in Mice.

Obesity is often associated with increased risk of cardiovascular diseases. Previous studies suggest that epidermal growth factor receptor (EGFR) antagonism may be effective for the treatment of angiotensin II-induced cardiac hypertrophy and diabetic cardiomyopathy. This study was performed to demonstrate if EGFR plays a role in the pathogenesis of /obesity-related cardiac injuries. The in vivo studies using both wild type (WT) and apolipoprotein E (ApoE) knockout mice fed with high fat diet (HFD) showed the beneficial effects of small-molecule EGFR inhibitors, AG1478 and 542, against obesity-induced myocardial injury. Administration of AG1478 and 542 significantly reduced myocardial inflammation, fibrosis, apoptosis, and dysfunction in both two obese mouse models. In vitro, EGFR signaling was blocked by either siRNA silencing or small-molecule EGFR inhibitors in (PA)-stimulated cardiomyocytes. EGFR inhibition attenuated PA-induced inflammatory response and apoptosis in H9C2 cells. Furthermore, we found that PA-induced EGFR activation was mediated by the upstream TLR4 and c-Src. This study has confirmed the detrimental effect of EGFR activation in the pathogenesis of obesity-induced cardiac inflammatory injuries in experimental mice, and has demonstrated the TLR4/c-Src-mediated mechanisms for PA-induced EGFR activation. Our data suggest that EGFR may be a therapeutic target for obesity-related cardiovascular diseases.

Keyword: hyperlipedemia

Effects of ammonium chloride, salicylate, and carnitine on oxidation in rat liver slices.

To explore the possible association of with hyperammonemia and aspirin ingestion, the effects of NH4+, salicylate, and carnitine on the oxidation of [1-14C] to -soluble products (ASP) and to CO2 were investigated in rat liver slices. DL-carnitine (5 mM) increased total oxidation (ASP + CO2) more than oxidation to CO2. KCN (1.5 mM) inhibited more than 90% of the oxidation. NH4Cl inhibited the oxidation that reached a maximum at about 40 mM, but the inhibition of oxidation to CO2 (63%) was larger than that of total oxidation (30%). Carnitine did not influence NH4+ inhibition, which is consistent with the results reported for isolated mitochondria. Salicylate effects depended on salicylate concentration as well as on the presence of carnitine. In the absence of carnitine, inhibition of total oxidation reached 90% at 3 mM salicylate but that of oxidation to CO2 reached 50%. Velocity calculated at saturating concentration for total oxidation was slightly increased by 0.75 mM salicylate, but the increase for oxidation to CO2 was larger. At 3 mM salicylate, velocity at saturating concentration for the oxidation was decreased, but the decrease for oxidation to CO2 was smaller than for total oxidation. Carnitine partially relieved the inhibition of total oxidation and further increased the formation of CO2. The combination of 20 mM NH4Cl and 0.75 mM salicylate inhibited total oxidation, which was more than additive of the individual effects, and carnitine partially relieved the inhibition. It is concluded that NH4+ exerted a stronger inhibition of oxidation to CO2 than of oxidation to ASP, whereas salicylate strongly inhibited the oxidation to ASP but increased the oxidation to CO2 by uncoupling mitochondrial oxidative phosphorylation. Therefore, hyperammonemia and aspirin ingestion can inhibit fatty oxidation and mitochondrial metabolism that could lead to the pathophysiology seen in some childhood diseases such as Reye\'s syndrome. Carnitine therapy might offer some benefits.

Keyword: hyperlipedemia

Astragalus polysaccharide suppresses skeletal muscle myostatin expression in diabetes: involvement of ROS-ERK and NF-κB pathways.

The antidiabetes drug astragalus polysaccharide (APS) is capable of increasing insulin sensitivity in skeletal muscle and improving whole-body glucose homeostasis. Recent studies suggest that skeletal muscle secreted growth factor myostatin plays an important role in regulating insulin signaling and insulin resistance. We hypothesized that regulation of skeletal muscle myostatin expression may be involved in the improvement of insulin sensitivity by APS.APS was administered to 13-week-old diabetic KKAy and nondiabetic C57BL/6J mice for 8 weeks. Complementary studies examined APS effects on the saturated palmitate-induced insulin resistance and myostatin expression in C2C12 cells.APS treatment ameliorated hyperglycemia, , and insulin resistance and decreased the elevation of myostatin expression and malondialdehyde production in skeletal muscle of noninsulin-dependent diabetic KKAy mice. In C2C12 cells in vitro, saturated palmitate-induced impaired glucose uptake, overproduction of ROS, activation of extracellular regulated protein kinases (ERK), and NF-κB were partially restored by APS treatment. The protective effects of APS were mimicked by ERK and NF-κB inhibitors, respectively.Our study demonstrates elevated myostatin expression in skeletal muscle of type 2 diabetic KKAy mice and in cultured C2C12 cells exposed to palmitate. APS is capable of improving insulin sensitivity and decreasing myostatin expression in skeletal muscle through downregulating ROS-ERK-NF-κB pathway.

Keyword: hyperlipedemia

Development of an in vitro model to study hepatitis C virus effects on hepatocellular lipotoxicity and lipid metabolism.

Hepatic steatosis is common in patients infected with hepatitis C virus (HCV). Particularly in patients infected with non-genotype 3 HCV, hepatic steatosis is closely related to factors of the metabolic syndrome such as . However, the molecular mechanisms involved in this "metabolic" steatosis in non-3 genotype HCV infections are not well understood. Here, we aimed to develop an in vitro model to study the effect of genotype 1 HCV infection on hepatic lipotoxicity and lipid metabolism. Cellular lipid accumulation was induced in Huh-7 hepatoma cells transfected with HCV genotype 1b replicon (HCV) by incubation with increasing doses of (C16:0) or oleic (C18:1 n-9) complexed to albumin mimicking hyperlipidemic conditions. Mock transfected hepatoma cells (HCV) were used as controls. Incubation with oleic concentrations as high as 0.5\u202fmM did not induce toxic effects in HCV or HCV cells. In contrast, incubation with caused dose-dependently cytotoxic effects which were more pronounced in HCV compared to HCV cells. Further analysis with subtoxic and oleic concentrations revealed a higher uptake of fatty acids and intracellular triglyceride accumulation in HCV compared to HCV cells. Carnitine palmitoyltransferase I (CPT1) expression, indicative of mitochondrial beta-oxidation, was markedly stimulated by lipid exposure in HCV but not in HCV cells. Furthermore, heme oxygenase 1 (HMOX1) expression levels increased in FA stimulated cells, and this increase was significantly higher in HCV compared to HCV cells. In contrast, expression of the key enzymes of hepatic de novo lipogenesis fatty synthase (FASN) and stearoyl-CoA desaturase (SCD-1) was significantly reduced upon oleate exposure in HCV but not in HCV cells. In summary, our newly developed cell culture model revealed effects of HCV genotype 1b infection on metabolic susceptibility to lipid accumulation and toxicity particularly to saturated lipids. These results may indicate that HCV (genotype 1b) infected individuals with may benefit from dietary or pharmacological intervention.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: hyperlipedemia

in the sn-2 position of triacylglycerols acutely influences postprandial lipid metabolism.

The triacylglycerol structure of saturated fats may influence postprandial lipemia.We tested the hypothesis that high-fat meals rich in (16:0) in the sn-2 position decrease lipemia.Postprandial changes in plasma lipids, apolipoprotein B48, and cytokines were compared in healthy men (n = 25) and women (n = 25) by using a randomized crossover design after meals that provided 50 g fat supplied as high-oleic sunflower oil (control), palm olein (PO), interesterified palm olein (IPO), and lard containing 0.6, 9.2, 39.1, and 70.5 mol% 16:0, respectively, at sn-2.The sn-2-rich meals elicited different postprandial responses in plasma concentrations of nonesterified fatty (meal × time, P = 0.00014), triacylglycerol (meal × time, P = 0.002), and apolipoprotein B48 (meal × time × sex, P = 0.008). Nonesterified fatty concentrations were lower up to 3 h after lard and IPO meals than after control or PO meals. Triacylglycerol increased less steeply after lard and IPO meals than after control and PO meals; the incremental AUCs (iAUCs) were 34% (95% CI: 7%, 124%; P < 0.05) and 26% (95% CI: 16%, 132%; P < 0.05) lower after lard than after control and PO meals, respectively. In men, the maximal increment in apolipoprotein B48 was 14% (95% CI: 3%, 25%; P < 0.05) and 16% (95% CI: 2%, 30%; P < 0.05) lower for lard and IPO, respectively, compared with control. The postprandial iAUC in triacylglycerol was 51% lower in women (P = 0.001) than in men. Plasma IL-6 increased postprandially, but IL-8, TNF-α, and E-selectin decreased after all meals.Fats with a higher proportion of in the sn-2 position decrease postprandial lipemia in healthy subjects. This trial was registered at controlled-trials.com as ISRCTN20774126.

Keyword: hyperlipedemia

-Induced Podocyte Apoptosis via the Reactive Oxygen Species-Dependent Mitochondrial Pathway.

Chronic kidney disease (CKD) is often accompanied by , which accelerates progression of the disease. Podocyte injury can lead to dysfunction of the glomerular filtration barrier, which is associated with proteinuria, a risk marker for the progression of CKD. Our previous studies demonstrated that (PA) can induce podocyte apoptosis; however, the underlying mechanisms are unclear. In the present study, we investigated the specific molecular mechanisms of PA-induced apoptosis in cultured podocytes.We cultured mouse podocytes and treated them with PA. Then, cell viability was measured using the Cell Counting Kit-8 colorimetric assay, lipid uptake was assessed by Oil Red O staining and boron-dipyrromethene staining, apoptosis was measured by flow cytometry, mitochondrial injury was assessed by JC-1 staining and transmission electron microscopy, and mitochondrial production of reactive oxygen species (ROS) was evaluated by fluorescence microscopy using the MitoSOX Red reagent. The effects of PA on the mitochondria-mediated caspase activation pathway were investigated by examining the expression of caspase-8, cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), B-cell lymphoma 2 (Bcl-2), Bax, Bid, cytochrome c, and Fas-associated protein with death domain (FADD) using western blotting. The translocation of Bax and cytochrome c were detected by immunofluorescence.PA treatment significantly increased lipid accumulation and induced podocyte apoptosis. We investigated whether the two primary apoptosis signaling pathways (death receptor-mediated pathway and mitochondria-mediated pathway) were involved in the execution of PA-induced podocyte apoptosis, and found that the levels of FADD, caspase-8, and Bid did not significantly change during this process. Meanwhile, PA treatment induced an increase in Bax protein expression and a decrease in Bcl-2 protein expression, with Bax translocation to the mitochondria. Furthermore, PA treatment induced mitochondrial impairment, and triggered the release of cytochrome c from the mitochondria to cytosol, with a concomitant dose-dependent increase in the levels of cleaved caspase-9, cleaved caspase-3, and PARP. Meanwhile, PA treatment increased mitochondrial production of ROS, and the mitochondria-targeted antioxidant mitoTEMPO significantly ameliorated PA-induced podocyte apoptosis.Our findings indicated that PA induced caspase-dependent podocyte apoptosis through the mitochondrial pathway, and mitochondrial ROS production participated in this process, thus potentially contributing to podocyte injury.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: hyperlipedemia

Small molecule kaempferol modulates PDX-1 protein expression and subsequently promotes pancreatic β-cell survival and function via CREB.

Chronic causes β-cell apoptosis and dysfunction, thereby contributing to the pathogenesis of type 2 diabetes (T2D). Thus, searching for agents to promote pancreatic β-cell survival and improve its function could be a promising strategy to prevent and treat T2D. We investigated the effects of kaempferol, a small molecule isolated from ginkgo biloba, on apoptosis and function of β-cells and further determined the mechanism underlying its actions. Kaempferol treatment promoted viability, inhibited apoptosis and reduced caspase-3 activity in INS-1E cells and human islets chronically exposed to palmitate. In addition, kaempferol prevented the lipotoxicity-induced down-regulation of antiapoptotic proteins Akt and Bcl-2. The cytoprotective effects of kaempferol were associated with improved insulin secretion, synthesis, and pancreatic and duodenal homeobox-1 (PDX-1) expression. Chronic significantly diminished cyclic adenosine monophosphate (cAMP) production, protein kinase A (PKA) activation, cAMP-responsive element binding protein (CREB) phosphorylation and its regulated transcriptional activity in β-cells, all of which were restored by kaempferol treatment. Disruption of CREB expression by transfection of CREB siRNA in INS-1E cells or adenoviral transfer of dominant-negative forms of CREB in human islets ablated kaempferol protection of β-cell apoptosis and dysfunction caused by palmitate. Incubation of INS-1E cells or human islets with kaempferol for 48h induced PDX-1 expression. This effect of kaempferol on PDX-1 expression was not shared by a host of structurally related flavonoid compounds. PDX-1 gene knockdown reduced kaempferol-stimulated cAMP generation and CREB activation in INS-1E cells. These findings demonstrate that kaempferol is a novel survivor factor for pancreatic β-cells via up-regulating the PDX-1/cAMP/PKA/CREB signaling cascade.Copyright © 2013 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Genetic obesity alters recruitment of TANK-binding kinase 1 and AKT into hypothalamic lipid rafts domains.

Lipid rafts (LRs) are membrane subdomains enriched in cholesterol, glycosphingolipids and sphingolipids containing saturated fatty . Signaling proteins become concentrated in these microdomains mainly by saturated fatty modification, thus facilitating formation of protein complexes and activation of specific signaling pathways. High intake of saturated fatty acids promotes inflammation and insulin resistance, in part by disrupting insulin signaling pathway. Here we investigate whether lipid-induced toxicity in obesity correlates with altered composition of insulin signaling proteins in LRs in the brain. Our results showed that insulin receptor (IR) is highly concentrated in LRs fraction in comparison with soluble or postsynaptic density (PSD) fractions. Analysis of LRs domains from hippocampus of obese mouse showed a significant decrease of IR and its downstream signaling protein AKT, while in the PSD fraction we detected partial decrease of AKT and no changes in the IR concentration. No changes were shown in the soluble extract. In hypothalamus, genetic obesity also decreases interaction of AKT, but we did not detect changes in the IR distribution. However, in this structure genetic obesity increases recruitment of the IR negative regulator TANK-binding kinase 1 (TBK1) into LRs and PSD fraction. No changes of AKT, IR and TBK1 were found in soluble fractions of obese in comparison with lean mice. In vitro studies showed that incubation with saturated but not with unsaturated docosahexaenoic (DHA) or palmitoleic decreases association of IR and AKT and increases TBK1 recruitment into LRs and PSD domains, emulating what happens in the obese mice. TBK1 recruitment to insoluble domains correlates with decreases of IR tyrosine phosphorylation and ser473 AKT phosphorylation, markers of insulin resistance. These data support the hypothesis that associated with genetic obesity alters targeting of TBK1 and insulin signaling proteins into insoluble LRs domains.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: hyperlipedemia

Hypolipidemic, antiobesity, and hypoglycemic-hypoinsulinemic effects of beta,beta\'-methyl-substituted hexadecanedioic in sand rats.

Treatment of male sand rats kept on a balanced laboratory chow diet ad libitum with beta,beta\'-tetramethyl-substituted hexadecanedioic (MEDICA 16) resulted in a hypolipidemic effect accompanied by an extensive reduction in adiposity, with a concomitant hypoglycemic-hypoinsulinemic effect. The overall effect was sustained as long as the drug was administered. The hypolipidemic effect of MEDICA 16 consisted of a 70 and 40% decrease in plasma triacylglycerols and cholesterol, respectively, and resulted from inhibition of liver lipogenesis and cholesterogenesis. Adipose reduction by MEDICA 16 treatment or calorie restriction consisted of a 75-90% decrease in the perirenal, omental, epididymal, and subcutaneous fat, with a 50% decrease in liver neutral lipids. The reduction in adiposity was accounted for by a respective decrease in the lipid content of individual adipocytes, with a concomitant decrease in the number of adipocytes of selected adipose tissues. The decrease induced in adiposity by MEDICA 16 treatment could not be accounted for by anorectic or cathartic effects of the drug. The hypoglycemic-hypoinsulinemic effect of MEDICA 16 consisted of amelioration of the tolerance of glucose with normalization of plasma insulin. It was accompanied by an eightfold increase in the number of insulin receptors in epididymal adipocytes, which was, however, counteracted by a decrease in their affinity for insulin. The receptor and postreceptor effects exerted by MEDICA 16 were similar to those of calorie restriction. The overall effect of MEDICA 16 in sand rats may reflect the pharmacological potential of MEDICA compounds in pathological hyperlipidemic-obesity-diabetic syndromes.

Keyword: hyperlipedemia

Chemical characterization of the lipophilic fraction of Livistona decipiens and Livistona chinensis fruit pulps (Palmae) and assessment of their anti-hyperlipidemic and anti-ulcer activities.

The oil of the dried pulps of Livistona decipiens and L. chinensis palm fruits have been studied for the first time by gas chromatography-mass spectrometry for their unsaponifiable matter (USM) and fatty composition (FAME). The antihyperlipidemic and anti-ulcer activities for both oils were also assayed. The principal fatty of L. decipiens pulp oil was oleic (53.4 %) and of L. chinensis pulp oil (47.4 %). In relation to anti-hyperlipidemic properties, the pulp oil of L. decipiens presented a better profile than that of L. chinensis, in comparison with the reference standard (simvastatin). In addition, both pulp oils showed high anti-ulcer activity using an indomethacin-induced ulceration technique in rat stomach. The relationship between the anti-hyperlipidemic, anti-ulcer and chemical composition of the pulp oils is also discussed.

Keyword: hyperlipedemia

Fatty composition of serum cholesterol esters, and erythrocyte and platelet membranes as indicators of long-term adherence to fat-modified diets.

Dietary adherence to four different fat-modified diets was examined in 160 subjects by determining the fatty composition of serum cholesterol esters (CEs) and erythrocyte (ER) and platelet (PT) membranes in addition to food records. Subjects were randomly assigned to one of the following diet groups: 1) high-sat--35/14:104 (% of energy from total/saturated:monounsaturated:polyunsaturated fatty acids in the actual diet) 2) AHA (American Heart Association) type--32/10:8:8 3) monoene-enriched--34/11:11:5, or 4) low-fat--30/12:8:3 for 6 mo. Decreases in the proportions of in CEs were found in the AHA-type and monoene-enriched-diet groups. An increased proportion of linoleic in CEs was found in the AHA-type group. The differences in the proportions of in CEs and linoleic and palmitoleic acids in PTs were significant in the AHA-type and monoene-enriched-diet groups compared with the high-sat group. An increase in alpha-linolenic in CEs was an indicator of the use of low erucic rapeseed oil, which was the main source of monoenes in the monoene-enriched-diet group.

Keyword: hyperlipedemia

ICER-1gamma overexpression drives palmitate-mediated connexin36 down-regulation in insulin-secreting cells.

Channels formed by the gap junction protein connexin36 (Cx36) contribute to the proper control of insulin secretion. We investigated the impact of chronic on Cx36 expression in pancreatic beta-cells. Prolonged exposure to the saturated free fatty palmitate reduced the expression of Cx36 in several insulin-secreting cell lines and isolated mouse islets. The effect of palmitate was fully blocked upon protein kinase A (PKA) inhibition by H89 and (Rp)-cAMP, indicating that the cAMP/PKA pathway is involved in the control of Cx36 expression. Palmitate treatment led to overexpression of the inducible cAMP early repressor (ICER-1gamma), which bound to a functional cAMP-response element located in the promoter of the CX36 gene. Inhibition of ICER-1gamma overexpression prevented the Cx36 decrease, as well as the palmitate-induced beta-cell secretory dysfunction. Finally, freshly isolated islets from mice undergoing a long term high fat diet expressed reduced Cx36 levels and increased ICER-1gamma levels. Taken together, these data demonstrate that chronic exposure to palmitate inhibits the Cx36 expression through PKA-mediated ICER-1gamma overexpression. This Cx36 down-regulation may contribute to the reduced glucose sensitivity and altered insulin secretion observed during the pre-diabetic stage and in the metabolic syndrome.

Keyword: hyperlipedemia

Development of insulin resistance in the JCR:LA-cp rat: role of triacylglycerols and effects of MEDICA 16.

The JCR:LA-cp rat develops an extreme obese/insulin-resistant syndrome such that by 12 weeks of age, there is no longer any insulin-mediated glucose turnover. At 4 weeks of age, obese and lean rats have essentially identical basal and insulin-mediated glucose uptake in skeletal muscle. By 8 weeks of age, however, the obese rats no longer exhibit such intake. Plasma insulin concentrations in the normal fed state show only small increases up to 4 weeks, with a rapid rise to a marked hyperinsulinemia thereafter, with an age at half-development of 5.5 weeks. Plasma triacylglycerol concentrations in fed obese rats are elevated at 3 weeks and rise rapidly thereafter. The triacylglycerol content of skeletal muscle is significantly elevated in the obese rats at 4 weeks of age. Histological examination of Oil Red O-stained muscle tissue and transmission electron microscopy shows the presence of intracellular lipid droplets. Treatment with the potent triacylglycerol-lowering agent MEDICA 16 (beta,beta\'-tetramethylhexadecanedioic ) from 6 weeks of age reduces plasma lipids markedly, but it reduces body weight and insulin resistance only modestly. In contrast, treatment with MEDICA 16 from the time of weaning at 3 weeks of age results in the normalization of food intake and body weight to over 8 weeks of age. The development of hyperinsulinemia is also delayed until 8.5 weeks of age, and insulin levels remain strongly reduced. Plasma triacylglycerol concentrations remain at the same level as in lean rats, and neither an elevated muscle triacylglycerol content nor intracellular lipid droplets are found at 4 weeks of age. The results indicate that insulin resistance develops in the young animals and is not directly due to a genetically determined defect in insulin metabolism. The mechanism of induction instead appears to be related to an exaggerated triacylglycerol metabolism.

Keyword: hyperlipedemia

Protective role of 20-OH ecdysone on lipid profile and tissue fatty changes in streptozotocin induced diabetic rats.

is an associated complication of diabetes mellitus. The association of hyperglycemia with an alteration of lipid parameters presents a major risk for cardiovascular complications in diabetes. The present study was designed to examine the antihyperlipidemic effect of 20-OH ecdysone on lipid profile and tissue fatty changes in streptozotocin induced diabetic rats. The levels of blood glucose, cholesterol, triglycerides, free fatty acids, phospholipids, low density lipoprotein, very low density lipoprotein, high density lipoprotein, lipoprotein lipase, lecithin cholesterol acyl transferase, 3-hydroxy 3-methylglutaryl coenzyme A reductase and fatty composition were estimated in plasma, liver and kidneys of control and experimental groups of rats. Oral administration of 20-OH ecdysone at a dose of 5mg/kg bodyweight per day to STZ-induced diabetic rats for a period of 30 days resulted in a significant reduction in fasting blood glucose, cholesterol, triglycerides, free fatty acids, phospholipids, low density lipoprotein, very low density lipoprotein, 3-hydroxy 3-methylglutaryl coenzyme A reductase and elevation of high density lipoprotein, lipoprotein lipase and lecithin cholesterol acyl transferasein comparison with diabetic untreated rats. Moreover, administration of 20-OH ecdysone to diabetic rats also decreased the concentrations of fatty acids, viz., , stearic (16:1) and oleic (18:1), whereas linolenic (18:3) and arachidonic (20:4) were elevated. The antihyperlipidemic effect of 20-OH ecdysone was compared with glibenclamide a well-known antihyperglycemic drug. The result of the present study indicates that 20-OH ecdysone showed an antihyperlipidemic effect in addition to its antidiabetic effect in experimental diabetes.Copyright © 2012 Elsevier B.V. All rights reserved.

Keyword: hyperlipedemia

Effect of troglitazone (CS-045) and bezafibrate on glucose tolerance, liver glycogen synthase activity, and beta-oxidation in fructose-fed rats.

To clarify the relationship between lipid and glucose metabolism abnormalities in fructose-fed rats, we examined whether an improvement of insulin sensitivity by troglitazone (CS-045) or a decrease in plasma lipids by bezafibrate affects the relationship between serum levels of lipid and glucose. In addition, we also examined changes in liver glycogen metabolism and beta-oxidation in fructose-fed rats. Troglitazone ameliorated fasting , hyperglycemia, and hyperinsulinemia. In addition, it augmented glycogen synthase activity by 53%, and decreased the mitochondrial beta-oxidation rate and ketone body production rate by 27% and 55%, respectively. However, hyperglycemia and liver glycogen synthase activity were not improved by bezafibrate treatment despite a marked reduction of serum triglyceride (TG) levels resulting from a 1.76-fold increase in mitochondrial oxidation and a 2.04-fold increase in hepatic ketone body production. These results suggest that abnormalities in glucose and lipid metabolism in fructose-fed rats, which are ameliorated by troglitazone, may be closely linked to reduced glycogen synthase activity in the liver.

Keyword: hyperlipedemia

Dietary saturated and trans fatty acids and lipoprotein metabolism.

Earlier studies have shown that not all saturated fatty acids are equally hypercholesterolaemic: stearic (C18:0) and saturated fatty acids with less than 12 carbon atoms are thought not to raise serum cholesterol levels. This suggests that the cholesterol-raising effects of saturated fatty acids can be attributed to lauric (C12:0), myristic (C14:0) and (C16:0). These three saturated fatty acids also have different effects on serum total cholesterol levels. Results from recent controlled dietary experiments suggest that lauric raises serum total and low-density lipoprotein (LDL) cholesterol levels slightly less, and myristic more, as compared with . Myristic , however, also causes higher levels of high-density lipoprotein (HDL) cholesterol. Stearic has only a slight effect on serum LDL and HDL cholesterol levels as compared with oleic . Trans monounsaturated fatty acids, however, increase LDL and decrease HDL cholesterol levels. Precise effects on lipoproteins of short and medium chain triglycerides (C4:0-C10:0) have never been examined.

Keyword: hyperlipedemia

The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis.

Hyperglycemia and (glycolipotoxicity)-triggered islet β-cell dysfunction is known to drive the progression of obesity-related type 2 diabetes, however the underlying mechanisms have not been clearly elucidated. The current study aimed to investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP-5) in islet cells under glucolipotoxic conditions. Using gene overexpression and knockdown approaches, we demonstrated that MKP-5 could alleviate glucolipotoxicity-induced apoptosis via the endoplasmic reticulum (ER) stress and mitochondrial apoptosis pathways owing to the altered regulation of caspase family members and ER stress-related molecules in MIN6 and primary islet cells. Overexpression of MKP-5 reversed the glucose and (GP)-induced impairment of insulin secretion as well as the abnormal decreases in the expression of islet functional genes, thereby maintaining the normal insulin secretory functionality, whereas the absence of MKP-5 aggravated islet cell dysfunction. In parallel, the production of ROS and increased inflammation-associated genes in response to GP were also reduced upon MKP-5 overexpression. Further, inhibition of JNK or P38 MAPK pathways resisted to glucolipotoxicity observed in MKP-5 knockdown MIN6 cells. These findings indicate that MKP-5 is an important mediator for glucolipotoxicity-induced islet cell dysfunction and apoptosis, with JNK and P38 as the critical downstream pathways.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

-induced activation of human T-lymphocytes and aortic endothelial cells with production of insulin receptors, reactive oxygen species, cytokines, and lipid peroxidation.

Diabetic conditions are associated with hyperglycemia and , but the role of saturated fatty acids (SFA) vs. unsaturated fatty acids (UFA) in activation of T-lymphocytes and human aortic endothelial cells (HAEC) is not known. We investigated in vitro effects of various concentrations of SFA (palmitate) and UFA (oleic, linoleic, linolenic, and arachidonic) acids in activation of these cells. These cells in presence of palmitate, but not UFA, exhibited time, and concentration-dependent emergence of insulin receptors, GLUT 4 expression, generation of ROS, cytokines, lipid peroxidation, and IRS-1. We conclude that both T-lymphocytes and HAEC share common characteristics in exhibiting activation of these cells to palmitate, but not to UFA, by developing insulin receptors and becoming insulin responsive tissues, a hitherto unknown response to palmitate. We hypothesize that these events may serve as protective defense mechanisms against acute effects of glucotoxicity and lipotoxicity in these cells.

Keyword: hyperlipedemia

Dietary fat saturation effects on low-density-lipoprotein concentrations and metabolism in various animal models.

Saturated vegetable oils (coconut, palm, and palm kernel oil) and fats (butter and lard) are hypercholesterolemic relative to monounsaturated and polyunsaturated vegetable oils. The increase in plasma low-density-lipoprotein-cholesterol (LDL-C) concentrations associated with consumption of saturated vegetable oils and fats is largely explained by a decrease in hepatic LDL receptor activity and an increase in the LDL-C production rate. Hepatic LDL receptor activity may be regulated by the messenger RNA concentration of the LDL receptor. The decrease in hepatic LDL receptor activity with saturated fat feeding is associated with decreased hepatic sterol O-acyltransferase activity and, therefore, a reduced inert pool of cholesteryl ester. A putative regulatory pool of cholesterol is increased with saturated fat feeding and suppresses LDL receptor activity, possibly through hepatic messenger RNA regulation. For most studies, an independent effect of a vegetable oil or fat could not be ascertained because there was no neutral control and at least two of the test oils or fats were varied. Animal data for the effects of individual fatty acids on plasma LDL-C concentrations and metabolism are sparse. The evidence suggests that caproic (6:0), caprylic (8:0), and capric (10:0) are neutral with respect to their LDL-C-raising properties and their ability to modulate LDL metabolism. Lauric (12:0), myristic (14:0), and (16:0) are approximately equivalent in their LDL-C-raising potential by reducing hepatic LDL receptor activity and increasing the LDL-C production rate, apparently via modulation of sterol O-acyltransferase activity. Stearic (18:0) appears to be neutral in its LDL-C-raising potential and how it affects LDL metabolism.

Keyword: hyperlipedemia

Low-density lipoprotein (LDL) oxidizability before and after LDL apheresis.

Oxidation of low-density lipoprotein (LDL) plays a major role in the development of atherosclerosis. Hypercholesterolemia has been associated with enhanced in vitro oxidation of LDL, and lipid-lowering therapy reduces LDL oxidizability. In the present study, we investigated whether LDL apheresis performed with different techniques affects in vitro diene formation (lag phase) and modification of apolipoprotein B-100 (apoB). Baseline and posttreatment diene formation was correlated with the baseline pattern of plasma total fatty acids. We then performed a computer-simulation study to test the hypothesis that LDL apheresis-induced changes in LDL oxidizability are related to changes in the mass ratio between freshly produced and older LDL. In 19 patients aged 49+/-7 years with heterozygous familial hypercholesterolemia (FH) regularly treated with either immunoadsorption, heparin-induced LDL precipitation (HELP), or dextran sulfate (DS) adsorption, we determined lipoprotein levels, the lag phase, apoB modification, and the fatty pattern in plasma samples drawn at the onset and termination of one LDL apheresis. LDL apheresis significantly decreased total cholesterol, high-density lipoprotein (HDL) cholesterol, LDL cholesterol, and triglycerides by 50.4%, 14.9%, 62.6%, and 33.6%, respectively. The lag phase increased by a significant mean of 9.8%; the charge of apoB was not altered. The lag phase before treatment positively correlated with the baseline concentration of plasma total , myristic, and oleic . The increase in the lag phase during treatment correlated with a high pretreatment concentration of lauric, linoleic, and docosahexanoic . The simulation study indicates that a temporary imbalance between two LDL compartments, one representing freshly secreted LDL and the other representing older LDL, could explain the observed increase in the lag phase after LDL apheresis. In conclusion, in patients with heterozygous FH, LDL apheresis performed with different techniques decreases the susceptibility of LDL to oxidation. This decrease may be related to a temporary mass imbalance between freshly produced and older LDL particles. Furthermore, the baseline fatty pattern influences pretreatment and posttreatment susceptibility to oxidation.

Keyword: hyperlipedemia

Mitochondria-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.

Can we use a mitochondrial-targeted antioxidant (Mitoquinone) during in vitro embryo culture to rescue developmental competence of oocytes matured under lipotoxic conditions, exhibiting mitochondrial dysfunction and oxidative stress?Supplementation of embryo culture media with Mitoquinone reduced oxidative stress and prevented mitochondrial uncoupling in embryos derived from metabolically compromised oocytes in vitro, leading to higher blastocyst rates and lower blastomeric apoptosis.Maternal metabolic disorders, such as obesity and type-II diabetes are associated with and elevated free fatty (FFA) concentrations in the ovarian follicular fluid (FF). Oocyte maturation under these lipotoxic conditions results in increased oxidative stress levels, mitochondrial dysfunction, reduced developmental competence and disappointing IVF results.A well-described bovine oocyte IVM model was used, where a pathophysiologically relevant elevated FF concentrations of (PA; 150\xa0μM or 300\xa0μM) were added to induce oxidative stress. After fertilization (Day 0, D0), zygotes were in vitro cultured (IVC, from D1 to D8) in standard fatty -free media in the presence or absence of Mitoquinone or its carrier triphenyl-phosphonium.Embryo cleavage and fragmentation (D2) and blastocyst rates (D8) were recorded. Mitochondrial activity and oxidative stress in cleaved embryos at D2 were determined using specific fluorogenic probes and confocal microscopy. D8 blastocysts were used to (i) examine the expression of marker genes related to mitochondrial unfolded protein responses (UPRmt; HSPD1 and HSPE1), mitochondrial biogenesis (TFAM), endoplasmic reticulum (ER) UPR (ATF4, ATF6 and BiP) and oxidative stress (CAT, GPX1 and SOD2) using real time RT-PCR; (ii) determine cell differentiation and apoptosis using CDX-2 and cleaved caspase-3 immunostaining; and (iii) measure mtDNA copy numbers. This was tested in a series of experiments with at least three independent replicates for each, using a total of 2525 oocytes. Differences were considered significant if a P value was <0.05 after Bonferroni correction.Exposure to PA during IVM followed by culture under control conditions resulted in a significant increase in oxidative stress in embryos at D2. This was associated with a significant reduction in mitochondrial inner membrane potential (uncoupling) compared with solvent control (P\xa0<\u20090.05). The magnitude of these effects was PA-concentration dependent. Consequently, development to the blastocysts stage was significantly hampered. Surviving blastocysts exhibited high apoptotic cell indices and upregulated mRNA expression indicating persistent oxidative stress, mitochondrial and ER UPRs. In contrast, supplementation of PA-derived zygotes with Mitoquinone during IVC (i) prevented mitochondrial uncoupling and alleviated oxidative stress at D2; and (ii) rescued blastocyst quality; normalized oxidative stress and UPR related genes and apoptotic cell indices (P\xa0>\u20090.01 compared with solvent control). Mitoquinone also improved blastocyst rate in PA-exposed groups, an effect that was dependent on PA concentration.N/A.This is a fundamental study performed using a bovine in vitro model using PA-induced lipotoxicity during oocyte maturation. PA is the most predominant FFA in the FF that is known to induce lipotoxicity; however, in vivo maturation in patients suffering from maternal metabolic disorders involve more factors that cannot be represented in one model. Nevertheless, focusing on the carryover oxidative stress as a known key factor affecting developmental competence, and considering the novel beneficial rescuing effects of Mitoquinone shown here, we believe this model is of high biological relevance.Human oocytes collected for IVF treatments from patients with maternal metabolic disorders are vulnerable to lipotoxicity and oxidative stress during in vivo maturation. The results shown here suggest that mitochondrial targeted therapy, such as using Mitoquinone, during IVC may rescue the developmental competence and quality of these compromised oocytes. After further clinical trials, this may be a valuable approach to increase IVF success rates for infertile patients experiencing metabolic disorders.This study was financially supported by a BOF/KP grant number 34399, from the University of Antwerp, Belgium. W.F.A.M. was supported by a postdoctoral fellowship from the Research Foundation-Flanders (FWO), grant number 12I1417N, Antwerp, Belgium. The Leica SP 8 confocal microscope used in this study was funded by the Hercules Foundation of the Flemish Government (Hercules grant AUHA.15.12). All authors have no financial or non-financial competing interests to declare.© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: hyperlipedemia

Nutritional and biochemical aspects of the hypolipidemic action of rice bran oil: a review.

In this paper, we review the effects of rice bran oil (RBO), an unconventional oil recently introduced onto the Indian market for human use. RBO contains oleic (38.4%), linoleic (34.4%), and linolenic (2.2%) as unsaturated fatty acids, and (21.5%) and stearic (2.9%) acids as saturated fatty acids. The unsaponifiable fraction (4.2%) has total tocopherols (81.3 mg%), gamma-oryzanol (1.6%), and squalene (320 mg%). Oryzanol is a mixture of ferulic esters of triterpene alcohols such as cycloartenol (CA) (106 mg%) and 24-methylene cycloartanol (494 mg%). Studies on experimental rats demonstrated a hypolipidemic effect of RBO. The unsaponifiable fraction of RBO lowers cholesterol levels. Feeding phytosterols, CA, and 24-methylene cycloartanol in amounts present in RBO to hypercholesterolemic rats for 8 weeks indicates that CA alone reduces cholesterol and triglyceride levels significantly. Endogenous sterol excretion increases in animals given CA. The accumulation of CA in the liver inhibits cholesterol esterase activity, which in turn leads to reduction in circulating cholesterol levels. CA is structurally similar to cholesterol and may compete with the binding sites of cholesterol and sequestrate cholesterol, which is metabolized to its derivatives. RBO, which is rich in tocopherols and tocotrienols, may improve oxidative stability. Tocotrienols inhibit HMG CoA reductase, resulting in hypocholesterolemia. The hypolipidemic effect of RBO has also been established in human subjects. Thus, RBO could be a suitable edible oil for patients with .

Keyword: hyperlipedemia

Postprandial lipid responses to an alpha-linolenic -rich oil, olive oil and butter in women: a randomized crossover trial.

Postprandial lipaemia varies with gender and the composition of dietary fat due to the partitioning of fatty acids between beta-oxidation and incorporation into triacylglycerols (TAGs). Increasing evidence highlights the importance of postprandial measurements to evaluate atherogenic risk. Postprandial effects of alpha-linolenic (ALA) in women are poorly characterized. We therefore studied the postprandial lipid response of women to an ALA-rich oil in comparison with olive oil and butter, and characterized the fatty composition of total lipids, TAGs, and non-esterified fatty acids (NEFAs) in plasma.A randomized crossover design (n = 19) was used to compare the postprandial effects of 3 meals containing 35 g fat. Blood samples were collected at regular intervals for 7 h. Statistical analysis was carried out with ANOVA (significant difference = P < 0.05).No significant difference was seen in incremental area under the curve (iAUC) plasma-TAG between the meals. ALA and oleic levels were significantly increased in plasma after ALA-rich oil and olive oil meals, respectively. was significantly increased in plasma-TAG after the butter meal. The ratios of 18:2 n-6 to18:3 n-3 in plasma-TAGs, three and seven hours after the ALA-rich oil meal, were 1.5 and 2.4, respectively. The corresponding values after the olive oil meal were: 13.8 and 16.9; and after the butter meal: 9.0 and 11.6.The postprandial p-TAG and NEFA response in healthy pre-menopausal women was not significantly different after the intake of an ALA-rich oil, olive oil and butter. The ALA-rich oil significantly affected different plasma lipid fractions and improved the ratio of n-6 to n-3 fatty acids several hours postprandially.

Keyword: hyperlipedemia

[Modification of pathologic fatty patterns in old age by combined clofibrinic and nicotinic treatment].

71 patients (42 men and 29 women, aged between 45 and 76) with primary hyperlipoproteinaemia (HLP) types IIa, IIb and IV, were treated with clofibric (Regadrin) and nicotinic derivatives (Radecol, Jupol) over a three-year period. Every four months, they underwent gas chromatographic analyses of the fatty spectrum (fa) in the fractions of cholesterol ester (Chol.E) and triglyceride (TGL) of the serum. The therapy resulted in an increase in the linoleic, linolenic, arachidonic, and eicosapentaenoic acids, and a fall in the , palmitoleic, stearic, oleic and eicosatrienic levels in the Chol.E fraction in the case of HLP types IIa and IIb, and in the TGL fraction in the case of HLP types IIb and IV. Selective competitive inhibition by unesterified fatty acids, blocked lipolysis, an impaired hepatogenic fatty- metabolism, an affected LCAT and an increased esterification of polyunsaturated fatty acids are discussed as possible mechanisms. The increase in polyunsaturated fatty acids and the decrease of saturated and monounsaturated fatty acids must be considered, as they are interrelated with the prostaglandin metabolism, to be a positive and vasoprotective effect which assumes special importance in middle and older age.

Keyword: hyperlipedemia

High-fat diet-dependent modulation of the delayed rectifier K(+) current in adult guinea pig atrial myocytes.

Obesity is associated with , electrical remodeling of the heart, and increased risk of supraventricular arrhythmias in both male and female patients. The delayed rectifier K(+) current (IK), is an important regulator of atrial repolarization. There is a paucity of studies on the functional role of IK in response to obesity. Here, we assessed the obesity-mediated functional modulation of IK in low-fat diet (LFD), and high-fat diet (HFD) fed adult guinea pigs. Guinea pigs were randomly divided into control and obese groups fed, ad libitum, with a LFD (10\xa0kcal% fat) or a HFD (45\xa0kcal% fat) respectively. Action potential duration (APD), and IK were studied in atrial myocytes and IKr and IKs in HEK293\xa0cells using whole-cell patch clamp electrophysiology. HFD guinea pigs displayed a significant increase in body weight, total cholesterol and total triglycerides within 50 days. Atrial APD at 30% (APD30) and 90% (APD90) repolarization were shorter, while atrial IK density was significantly increased in HFD guinea pigs. Exposure to (PA) increased heterologously expressed IKr and IKs densities, while oleic (OA), severely reduced IKr and had no effect on IKs. The data are first to show that in obese guinea pigs abbreviated APD is due to increased IK density likely through elevations of PA. Our findings may have crucial implications for targeted treatment options for obesity-related arrhythmias.Published by Elsevier Inc.

Keyword: hyperlipedemia

[The clinical biochemistry of hyperlipemia and hyperglycemia. Insulin and metabolism of fatty acids. Hypoglycemic effect of hyperlipemicpharmaceuticals].

The regulation of metabolism of glucose is billions years older than system of insulin and biological function of locomotion (function of motion). Hence hypoglycemic effect of hormone is mediated by alteration of metabolism of fatty acids. The insulin in physiological way deprives mitochondrions a possibility to metabolize ketone bodies, short chain, medium chain and long chain fatty acids and \'forces" them to oxidize glucose which phylogenetically is not an optimal substrate. The relationships between fatty acids and glucose in the Rendle cycle have an effect only on autocrine level (in cell) determining alternation of biological reactions of exotrophia (after food intake) and endotrophia (beyond food intake) in biological function of alimentation (trophology). The most anti-diabetic pharmaceuticals are as insulin hyperlipemic by their mechanism of action. The decrease content of lipid substrates of oxidation in cytosol of cells and mitochondrions "are forced" to oxidize glucose. In these conditions, insulin enhances absorption of glucose by cells through glucose carriers--GLUT4. The derivatives of sulfonil-urea increase secretion of insulin by beta-cells of islets. The biguanidines bond in cytosol covalently and irreversibly ketone bodies taking them away from oxidation in mitochondrions. The fibrates, glitazones, flavonoids and flavones, lipoic tio-fatty acids. The endogenous eicosanoids, derivatives omega-3 and omega-6 of essential polyolefinic fatty acids and conjugated unsaturated fatty acids are the antagonists of receptors of activation of proliferation of peroxisomes. In peroxisomes, they enhance alpha-, beta- and omega-oxidation of all exogenous a physiological fatty acids and excess of saturated fatty forming hypolipidemia in cytozol. The hypolipidemic pharmaceuticals with effect of beta-blocker of oxidation stop absorption of fatty acids by mitochondrions. The omega-3 essential polvolefinic fatty acids, simultaneously with hypolipidemic effect, activate function of GLUT4. In patients of middle age, the diabetes mellitus type II is a symptom of syndrome of atherosclerosis. The reason is that in cells the deficiency of essential polyolefinic fatty acids and is determined by derangement of synthesis of phospholipids and function of GLUT4. It is valid to consider diabetes mellitus primarily as a pathology of metabolism of fatty acids and secondly as a pathology of content of glucose. It is necessary to take into account both under treatment (tactic activities) and strategic program of prevention of diabetes mellitus in population.

Keyword: hyperlipedemia

aggravates inflammation of pancreatic acinar cells by enhancing unfolded protein response induced CCAAT-enhancer-binding protein β-CCAAT-enhancer-binding protein α activation.

Hypertriglyceridemia is an independent risk factor for acute pancreatitis, in which the pathological mechanisms are not fully illustrated. Intracellular inflammatory response is a key pathological response in acute pancreatitis and endoplasmic reticulum stress has been suggested to induce inflammation and CCAAT-enhancer-binding protein expression. Therefore, the current study aims to elucidate the possible relationship between endoplasmic reticulum stress and inflammation in hypertriglyceridemia associated pancreatitis and the possible involvement of CCAAT-enhancer-binding protein. In cholecystokinin-8 stimulated rat primary acinar cells, incubation with caused the activation of endoplasmic reticulum stress and inflammatory responses. Pre-incubation with the chemical chaperone 4-phenylbutyric inhibited inflammatory responses induced by , whereas stimulation with the endoplasmic reticulum stress inducer thapsigargin alone induced inflammatory responses. Meanwhile we found that the transcription factors CCAAT-enhancer-binding protein α and CCAAT-enhancer-binding protein β were also induced in the -stimulated pancreatic acinar cells, and were similarly inhibited by 4-phenylbutyric pre-incubation and induced by thapsigargin stimulation alone, indicating that endoplasmic reticulum stress was responsible for CCAAT-enhancer-binding protein α and CCAAT-enhancer-binding protein β induction in the pancreatic acinar cells. Knockdown of CCAAT-enhancer-binding protein β by siRNA transfection inhibited inflammatory responses and CCAAT-enhancer-binding protein α induction but did not affect endoplasmic reticulum stress. Our study provides strong evidence that in response to stimulation, endoplasmic reticulum stress induces inflammatory responses in pancreatic acinar cells through induction of the CCAAT-enhancer-binding protein family, wherein CCAAT-enhancer-binding protein β activation is responsible for CCAAT-enhancer-binding protein α activation.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: hyperlipedemia

Comparative studies of triacylglycerol structure of very low density lipoproteins and chylomicrons of normolipemic subjects and patients with type II hyperlipoproteinemia.

The triacylglycerols of very low density lipoproteins (VLDL) and of chylomicrons were analyzed in the fasting and postabsorptive states from normolipemic subjects and patients with Frederickson\'s Type II hyperlipoproteinemia, who subsisted on free choice diets, standard diets excluding lard, or were given a breakfast enriched in lard. The VLDL and chylomicrons were obtained by conventional ultracentrifugation, and the triacylglycerols were isolated by thin-layer chromatography (TLC). Representative sn-1,2-, sn-2-3- and sn-1,3-diacylglycerols were generated by partial Grignard degradation of the triacylglycerols and a stereospecific hydrolysis by phospholipase C of the mixed sn-1,2(2,3)-diacyl phosphatidylcholines prepared as intermediates. Representative sn-2-acylglycerols were obtained by hydrolysis with pancreatic lipase. Positional distribution of the fatty acids was established by subtracting in turn the fatty composition of the sn-2-position from the fatty composition of the sn-1,2- and sn-2,3-diacylglycerols. The molecular association of the fatty acids in the diacylglycerol moieties was determined by gas-liquid chromatography with mass spectrometry (GC/MS) of the tertiary-butyldimethylsilyl (t-BDMS) ethers. The molecular association of the fatty acids in the triacylglycerols was determined by 1-random 2-random 3-random calculation following experimental validation of the distribution. The results confirm a marked asymmetry in the positional distribution of the fatty acids in all triacylglycerol samples, with the predominantly in the sn-1-position, the unsaturated acids about equally divided between the sn-2- and sn-3-positions, and the stearic divided about equally between the sn-1- and sn-3-positions. The overall structure of the VLDL and chylomicron triacylglycerols from patients and control subjects was characterized by a non-correlative distribution of fatty acids under all dietary conditions.

Keyword: hyperlipedemia

5-HT 2 receptor mediates high-fat diet-induced hepatic steatosis and very low density lipoprotein overproduction in rats.

5-HT has been shown to mediate abnormality of hepatic lipid metabolism through activation of mammalian target of rapamycin (mTOR). However, it is unclear whether 5-HT is directly involved in high-fat diet (HFD)-induced hepatic steatosis.Male rats were allocated into seven groups with control, either HFD feeding, 5-HT treatment, or HFD feeding and 5-HT treatment with or without sarpogrelate treatment, all of which were executed for 4 weeks. HepG2 cells were exposed to 5-HT or (PA) with or without rapamycin or Sar treatment.Rats fed with HFD or exposed to 5-HT led to abnormalities with activated hepatic mTOR-S6K pathway, overproduction of hepatic triglycerides and VLDL with steatosis, and , which were exacerbated by a combination of HFD and 5-HT. Sarpogrelate significantly inhibited above abnormalities induced by HFD and 5-HT, alone or in a combination. Additionally, HFD caused up-regulation of 5-HT2 receptors (5-HTR), including 5-HTR and 5-HTR, and 5-HT synthesis in the liver, without obvious influence on other 5-HT receptors gene expression. In HepG2 cells, both PA and 5-HT induced overproduction of triglycerides and VLDL with lipid droplets, and PA up-regulated 5-HTR and 5-HTR expression and 5-HT synthesis as well. Rapamycin fully abolished PA or 5-HT-induced mTOR activation, which was more effective than sarpogrelate. However, the inhibitory effects of rapamycin on PA or 5-HT-induced overproduction of triglycerides and VLDL were less than sarpogrelate.Up-regulation of hepatic 5-HTR and 5-HT synthesis by HFD is crucial for HFD-induced overproduction of hepatic triglycerides and VLDL with .Copyright © 2016 Asia Oceania Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.

Keyword: hyperlipedemia

Identification of linoleic and arachidonic acids as the factors in hyperlipemic blood that increase [3H]thymidine incorporation in hepatoma 7288CTC perfused in situ.

Tumor growth and the incorporation of [3H]thymidine into tumor DNA in vivo are increased about 3 times in adult rats (greater than 250 g) after 1 to 2 days of starvation or the induction of diabetes with streptozotocin. These tumor growth responses require hyperlipemia and are reversed by refeeding or insulin treatment, respectively. They do not occur in young tumor-bearing rats (less than about 150 g) that lack appreciable fat stores. A direct relationship between the increased rates of both [3H]thymidine incorporation and tumor growth and host hyperlipemia suggests that tumor cell renewal in vivo in fed rats is limited by substances that are present in hyperlipemic blood. In this study we used a procedure for perfusion of solid tumors in situ to measure the sensitivity of tumor [3H]thymidine incorporation to hyperlipemic blood and to identify the rate-limiting substances. Tissue-isolated Morris hepatomas (7288CTC) growing in young or adult Buffalo rats were perfused with blood from donor rats. Hyperlipemic blood for perfusion was obtained from 2-day starved tumor-bearing (Buffalo) or non-tumor-bearing (Buffalo or Lewis) rats. At the end of the perfusions the tumors were labeled with a pulse of [3H]thymidine (2 microCi/g estimated tumor wet weight). [3H]Thymidine incorporation in tumors growing in fed adult rats was increased from 80 +/- 5 (SD) dpm/micrograms DNA at zero time (before perfusion) to 209 +/- 9 dpm/micrograms DNA (n = 3) after perfusion for 3 h. Tumors growing in fed or starved young rats showed similar responses, and hyperlipemic blood from non-tumor-bearing rats was as effective as hyperlipemic blood from tumor-bearing rats. Perfusion of tumors growing in starved rats with normolipemic blood from fed adult rats decreased [3H]thymidine incorporation from 211 +/- 13 dpm/micrograms DNA before perfusion to 68 +/- 9 dpm/micrograms DNA (n = 3) after perfusion for 3 h. Cells, plasma, and plasma subfractions from hyperlipemic blood were reconstituted to whole blood using plasma, cells, and whole blood, respectively, from fed rats and the mixtures were perfused into tumors growing in fed adult rats. Mixtures containing hyperlipemic plasma, lipid extracts (ethanol:acetone, 1:1) of hyperlipemic plasma, or albumin from hyperlipemic plasma increased tumor [3H]thymidine incorporation. Free fatty concentrations were increased about five times in hyperlipemic plasma and perfusion of tumors with normolipemic blood containing added linoleic and arachidonic acids increased [3H]thymidine incorporation. Blood mixtures containing , stearic, and oleic acids were inactive.(ABSTRACT TRUNCATED AT 400 WORDS).

Keyword: hyperlipedemia

Distinctive roles of unsaturated and saturated fatty acids in hyperlipidemic pancreatitis.

To investigate how the saturated and unsaturated fatty composition influences the susceptibility of developing acute pancreatitis.Primary pancreatic acinar cells were treated with low and high concentrations of different saturated and unsaturated fatty acids, and changes in the cytosolic Ca(2+) signal and the expression of protein kinase C (PKC) were measured after treatment.Unsaturated fatty acids at high concentrations, including oleic , linoleic , palmitoleic , docosahexaenoic , and arachidonic , induced a persistent rise in cytosolic Ca(2+) concentrations in acinar cells. Unsaturated fatty acids at low concentrations and saturated fatty acids, including , stearic , and triglycerides, at low and high concentrations were unable to induce a rise in Ca(2+) concentrations in acinar cells. Unsaturated fatty acids at high concentrations but not saturated fatty acids induced intra-acinar cell trypsin activation and cell damage and increased PKC expression.At sufficiently high concentrations, unsaturated fatty acids were able to induce acinar cells injury and promote the development of pancreatitis. Unsaturated fatty acids may play a distinctive role in the pathogenesis of pancreatitis through the activation of PKC family members.

Keyword: hyperlipedemia

High-density lipoprotein ameliorates -induced lipotoxicity and oxidative dysfunction in H9c2 cardiomyoblast cells via ROS suppression.

High levels circulating saturated fatty acids are associated with diabetes, obesity and . In heart, the accumulation of saturated fatty acids has been determined to play a role in the development of heart failure and diabetic cardiomyopathy. High-density lipoprotein (HDL) has been reported to possess key atheroprotective biological properties, including cellular cholesterol efflux capacity, anti-oxidative and anti-inflammatory activities. However, the underlying mechanisms are still largely unknown. Therefore, the aim of the present study is to test whether HDL could protect (PA)-induced cardiomyocyte injury and explore the possible mechanisms.H9c2 cells were pretreated with HDL (50-100\u2009μg/ml) for 2\u2009h followed by PA (0.5\u2009mM) for indicated time period. Our results showed that HDL inhibited PA-induced cell death in a dose-dependent manner. Moreover, HDL rescued PA-induced ROS generation and the phosphorylation of JNK which in turn activated NF-κB-mediated inflammatory proteins expressions. We also found that PA impaired the balance of BCL family proteins, destabilized mitochondrial membrane potential, and triggered subsequent cytochrome c release into the cytosol and activation of caspase 3. These detrimental effects were ameliorated by HDL treatment.PA-induced ROS accumulation and results in cardiomyocyte apoptosis and inflammation. However, HDL attenuated PA-induced lipotoxicity and oxidative dysfunction via ROS suppression. These results may provide insight into a possible molecular mechanism underlying HDL suppression of the free fatty -induced cardiomyocyte apoptosis.

Keyword: hyperlipedemia

Influence of linoleic on desaturation and uptake of deuterium-labeled and stearic acids in humans.

Objectives of this study were to investigate the desaturation of stearic (18:0) and (16:0), to determine if differences in their metabolism provide a reasonable explantation for differences in their effect on serum cholesterol levels, and to investigate the affect of linoleic on delta 9-desaturase products in man. Deuterium-labeled 16:0 and 18:0 were used to follow the metabolism of these fatty acids in young adult male subjects that were pre-fed diets containing two different levels of linoleic . Results indicate that absorption of 16:0 and 18:0 was similar when all components of the mixture used to formulate the deuterated fat mixture were kept above the melting point of tristearin. The percent of 18:0 desaturated to 9c-18:1 was higher than the percent of 16:0 desaturated to 9c-16:1 (9.2% vs. 3.9%). The subject-to-subject variability suggests that differences in ability to desaturate saturated fatty acids may be related to the variability observed in response of serum cholesterol levels to dietary saturated fatty acids. Data for the distribution of 16:0 and 18:0 between triacylglycerol and phosphatidylcholine (PC) was markedly different. Based on PC data, phospholipid acyltransferase selectivity was about 2-fold higher for 18:0 than for 16:0. A 2-fold difference in the linoleic content of the pre-fed diets had little influence on desaturation or distribution of 16:0 and 18:0 between plasma lipid classes. A deuterium isotope effect was estimated to reduce delta 9-desaturase enzyme activity by 30-50%.

Keyword: hyperlipedemia

Serum and erythrocyte membrane phospholipids fatty composition in : effects of dietary intervention and combined diet and fibrate therapy.

is found to be associated with changes in fatty (FA) profiles. The aim of this study was to investigate the effects of AHA-Step-1 dietary treatment and combination of fibrates (gemfibrozil) with dietary intervention on serum and erythrocyte phospholipid FA composition in human . 78 study participants with were divided in two groups. In D group (n = 41) subjects followed AHA-Step-1 diet (<30% of total from fat, <10% of energy from saturated fat, and <300 mg cholesterol per day). D+F group (n = 37) followed Step-1 diet and were receiving gemfibrozil (300 mg/twice per day). Serum lipid levels and phospholipid serum and erythrocyte FA compositions were analyzed at the beginning and after 12 weeks of treatment. Alteration in serum and erythrocyte phospholipid FA profile were found in both groups. After both treatments we found significantly higher serum phospholipid percentages of n-3, n-6 and total polyunsaturated FA. Linoleic (LA, n-6) and docosahexaenoic (DHA, n-3) were higher in D group, but arachidonic (AA, n-6) and linolenic (LNA, n-3) in D+F group. In erythrocyte phospholipid levels of stearic, palmitoleic (16 : 1, n-7) and LA were significantly higher in D group, but , AA and eicosapentaenoic (EPA, n-3) in D+F group. Stronger correlation between serum triglycerides with EPA and DHA in erythrocyte membrane phospholipid was found in D+F group. Markedly increased percentage of AA in serum and erythrocyte membrane phospholipid in hyperlipidemic patients receiving gemfibrozil on Step-1 diet is especially important for physiological functions (inflammation, vascular tone, hemostasis etc.) in relation to cardiometabolic risk.

Keyword: hyperlipedemia

[Prevention of atherosclerosis. Excess of in food--a cause of hypercholesterolemia, inflammatory syndrome, insulin resistance in myocytes, and apoptosis].

Unity of the pathogenesis of atherosclerosis, type 2 diabetes mellitus, and metabolic syndrome gives rise to impaired biological function of adaptation, altered biological function of exotrophy (external feeding) and endoecology ("purity" of the intercellular medium). Biological reactions of inflammation and hydrodynamic pressure, or blood pressure, are in vivo activated to compensate for intercellular debris accumulation by endogenous phlogogens--ligand-free low density lipoproteins (LDL). The biological reactions jointly remove LDL from blood to the intima of elastic type arteries, to interstitial tissue for the local pool of the intravascular medium. The causes of formation of aphysiological LDLs are a preponderance of palmitate-oleate-palmitate triglycerides in the latter and impaired hydrolysis upon exposure to post-heparin lipase to give rise to small, dense LDLs; intimal macrophages utilize the debris only partially and develop atheromatosis from polyenic fatty acids (FA) etherified by cholesterol alcohol. Excess of saturated fatty (sFA) is responsible for the lowered permeability of the plasma membrane, cell death via the mechanism similar to apoptosis. Aphysiological protein palmitoylation (covalent interaction with sFA) increased the debris accumulation of the intercellular medium and the activity of both biological reactions. Elevated plasma sFA and its enhanced passive absorption in the form of unetherified FA, as well as high C-reactive protein levels are a cause of insulin resistance. The only way to prevent atherosclerosis in the population is to normalize the biological function of exotrophy when the energy value ratio of FA, proteins and carbohydrates is 1:1:1 and that of sFA, monoenic, and polyenic FA is also 1:1:1. The lower amount of sFA and the higher concentration of essential polyenic FA, the lower blood levels of cholesterol alcohol and triglycerides are. At the same time, simultaneously activations and the biological function of locomotion are a level of physical activity.

Keyword: hyperlipedemia

High Fat Diet Upregulates Fatty Oxidation and Ketogenesis via Intervention of PPAR-γ.

Systemic and intracellular lipid accumulation induced by chronic high fat diet (HFD) leads to enhanced fatty oxidation (FAO) and ketogenesis. The present study was aimed to determine whether activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) by surplus free fatty acids (FA) in hyperlipidemic condition, has a positive feedback regulation over FAO and ketogenic enzymes controlling lipotoxicity and cardiac apoptosis.8 weeks old C57BL/6 wild type (WT) or PPAR-γ-/- mice were challenged with 16 weeks 60% HFD to induce obesity mediated type 2 diabetes mellitus (T2DM) and diabetic cardiomyopathy. Treatment course was followed by echocardiographic measurements, glycemic and lipid profiling, immunoblot, qPCR and immunohistochemistry (IHC) analysis of PPAR-γ and following mitochondrial metabolic enzymes 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), mitochondrial β- hydroxy butyrate dehydrogenase (BDH1) and pyruvate dehydrogenase kinase isoform 4 (PDK4). In vivo model was translated in vitro, with neonatal rat cardiomyocytes (NRCM) treated with PPAR-γ agonist/antagonist and PPAR-γ overexpression adenovirus in presence of (PA). Apoptosis was determined in vivo from left ventricular heart by TUNEL assay and immunoblot analysis.We found exaggerated circulating ketone bodies production and expressions of the related mitochondrial enzymes HMGCS2, BDH1 and PDK4 in HFD-induced diabetic hearts and in PA-treated NRCM. As a mechanistic approach we found HFD mediated activation of PPAR-γ is associated with the above-mentioned mitochondrial enzymes. HFD-fed PPAR-γ-/-mice display decreased hyperglycemia, associated with increased insulin responsiveness as compared to HFD-fed WT mice PPAR-γ-/-HFD mice demonstrated a more robust functional recovery after diabetes induction, as well as significantly reduced myocyte apoptosis and improved cardiac function.PPAR-γ has been described previously to regulate lipid metabolism and adipogenesis. The present study suggests for the first time that increased PPAR-γ expression by HFD is responsible for cardiac dysfunction via upregulation of mitochondrial enzymes HMGCS2, BDH1 and PDK4. Targeting PPAR-γ and its downstream mitochondrial enzymes will provide novel strategies in preventing metabolic and myocardial dysfunction in diabetes mellitus.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: hyperlipedemia

Chokeberry Juice Containing Polyphenols Does Not Affect Cholesterol or Blood Pressure but Modifies the Composition of Plasma Phospholipids Fatty Acids in Individuals at Cardiovascular Risk.

Chokeberry polyphenols have been suggested to reduce cholesterol and blood pressure and thus protect against cardiovascular diseases (CVD), but the evidence in humans is limited and inconsistent. This randomized double-blinded three-parallel groups trial investigated the changes in various anthropometric and clinical biomarkers, and in plasma phospholipids fatty acids (PPFA) in volunteers at cardiovascular risk after a four-week intervention with 100 mL/day of (1) chokeberry juice with a high-dose of polyphenols (1177.11 mg gallic equivalents, GAE); (2) chokeberry juice with a low-dose of polyphenols (294.28 mg GAE) and; (3) a nutritionally matched polyphenol-free placebo drink. Our results indicate that the intake of chokeberry juice containing either the low or the high dose of polyphenols cannot be linked with a reduction in total- and low-density lipoprotein (LDL)cholesterol or in systolic (SBP) and diastolic (DBP) blood pressure in comparison with the consumption of the placebo drink. However, we found evidence of moderate changes in the PPFA, i.e., increased saturated fatty acids (SFA), mostly , and reduced n-6 polyunsaturated fatty acids (PUFA), principally linoleic (LA) with the intake of chokeberry against the placebo. These effects may be associated with the polyphenols but we could not differentiate a clear dose-response effect. Further research is still needed to elucidate the contribution of the polyphenolic fraction to the potential cardiovascular effects of the chokeberry and to build up the evidence of its potential benefit via the modulation of PPFA composition.

Keyword: hyperlipedemia

Hydrogen Sulphide modulating mitochondrial morphology to promote mitophagy in endothelial cells under high-glucose and high-palmitate.

Endothelial cell dysfunction is one of the main reasons for type II diabetes vascular complications. Hydrogen sulphide (H S) has antioxidative effect, but its regulation on mitochondrial dynamics and mitophagy in aortic endothelial cells under hyperglycaemia and is unclear. Rat aortic endothelial cells (RAECs) were treated with 40 mM glucose and 200 μM palmitate to imitate endothelium under hyperglycaemia and , and 100 μM NaHS was used as an exogenous H S donor. Firstly, we demonstrated that high glucose and palmitate decreased H S production and CSE expression in RAECs. Then, the antioxidative effect of H S was proved in RAECs under high glucose and palmitate to reduce mitochondrial ROS level. We also showed that exogenous H S inhibited mitochondrial apoptosis in RAECs under high glucose and palmitate. Using Mito Tracker and transmission electron microscopy assay, we revealed that exogenous H S decreased mitochondrial fragments and significantly reduced the expression of p-Drp-1/Drp-1 and Fis1 compared to high-glucose and high-palmitate group, whereas it increased mitophagy by transmission electron microscopy assay. We demonstrated that exogenous H S facilitated Parkin recruited by PINK1 by immunoprecipitation and immunostaining assays and then ubiquitylated mitofusin 2 (Mfn2), which illuminated the mechanism of exogenous H S on mitophagy. Parkin siRNA suppressed the expression of Mfn2, Nix and LC3B, which revealed that it eliminated mitophagy. In summary, exogenous H S could protect RAECs against apoptosis under high glucose and palmitate by suppressing oxidative stress, decreasing mitochondrial fragments and promoting mitophagy. Based on these results, we proposed a new mechanism of H S on protecting endothelium, which might provide a new strategy for type II diabetes vascular complication.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: hyperlipedemia

The effect of omega-3 carboxylic acids on apolipoprotein CIII-containing lipoproteins in severe hypertriglyceridemia.

Lipoprotein subspecies containing apoCIII adversely affect cardiovascular disease (CVD) risk; for example, low density lipoprotein (LDL) with apoCIII is a stronger CVD predictor than LDL without apoCIII. The Epanova for Lowering Very High Triglycerides (EVOLVE) trial showed that Epanova (omega-3 carboxylic acids [OM3-CA]) significantly lowered TG and apoCIII but raised LDL-C. However, it is unknown what subspecies of LDL were affected by treatment.To determine how lipoprotein subspecies are affected by omega-3 fatty treatment, we studied the effect of OM3-CA on apoCIII concentrations in high density lipoprotein (HDL), LDL, and very low density lipoprotein (VLDL) and on the concentrations of subspecies of HDL, LDL, and VLDL that contain or do not contain apoCIII.We analyzed plasma from a subset of subjects from the EVOLVE trial, a 12-week double-blind study of 399 subjects with fasting TG of 500 to 2000\xa0mg/dL who were randomized to OM3-CA 2, 3, or 4\xa0g/d or olive oil (placebo).OM3-CA significantly reduced plasma apoCIII relative to placebo, as well as apoCIII in HDL, and apoCIII in LDL. Treatment did not significantly affect the concentration of LDL with apoCIII, a subspecies highly associated with CVD risk. OM3-CA increased selectively the concentration of LDL that does not contain apoCIII, a subspecies with a weak relation to coronary heart disease. The reduction in apoCIII was associated with plasma increases in eicosapentaenoic , docosahexaenoic , and arachidonic and decreases in linoleic, , and oleic acids.Reduction in apoCIII may be a mechanism for the TG-lowering effects of OM3-CA. The increase in LDL-C seen in the EVOLVE trial may not be associated with increased risk of CVD.Copyright © 2016 National Lipid Association. Published by Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Hypolipidemic effect of beta, beta\'-tetramethyl hexadecanedioic (MEDICA 16) in hyperlipidemic JCR:LA-corpulent rats.

Short-term treatment of male and female obese JCR:LA-corpulent rats with beta,beta\'-tetramethyl hexadecanedioic (MEDICA 16) resulted in a marked decrease (as much as 80%) in plasma triglyceride values, with a concomitant decrease in the highly elevated very low density lipoprotein (VLDL) levels of the corpulent rat. There were modest decreases in cholesterol levels and increases in low density lipoprotein and high density lipoprotein lipids. The concentrations of apolipoproteins C-II and C-III were decreased in both the whole-serum and the VLDL fractions. Food consumption, rate of weight gain, fasting insulin levels, and the integrated insulin response to an intravenous glucose load remained unaffected. The decrease in plasma VLDL may be accounted for by inhibition of liver long-chain fatty synthesis at the level of ATP citrate lyase, with a concomitant reduction of VLDL triglyceride production by the liver. This decrease in plasma VLDL production was accompanied by a twofold to threefold increase in the triglyceride and cholesterol components of the low density lipoprotein and high density lipoprotein fractions, together with a twofold to fourfold decrease in plasma apolipoprotein, indicating that activation of plasma VLDL catabolism may further account for the overall hypolipidemic effect induced by MEDICA 16. The overall hypolipidemic effect of MEDICA 16 may be expected to inhibit the spontaneous atherogenic sequelae induced in the corpulent rat by severe VLDL .

Keyword: hyperlipedemia

Clustering effects on postprandial insulin secretion and sensitivity in response to meals with different fatty compositions.

Dietary fatty acids play a role in glucose homeostasis. The aim of this study was to assess the individual relationship between dietary saturated (SFA), monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids with postprandial β-cell function and insulin sensitivity in subjects with normal and high fasting triglycerides. We assessed postprandial β-cell function (by the insulinogenic index and the ratio of the insulin to glucose areas under the time-concentration curve) and insulin sensitivity (by the oral glucose and the minimal model insulin sensitivity indices) over four nonconsecutive, randomly assigned, high-fat meals containing a panel of SFA ( and stearic acids), MUFA (palmitoleic and oleic acids) and PUFA (linoleic and α-linolenic acids) in 14 subjects with normal and in 14 subjects with high fasting triglycerides. The proportions of each fatty in the meals and the values for surrogate measures of postprandial β-cell function and insulin sensitivity were subjected to a Pearson correlation and hierarchical cluster analysis, which revealed two classes of dietary fatty acids for regulating postprandial glucose homeostasis. We successfully discriminated the adverse effects of SFA from the beneficial effects of MUFA oleic on postprandial β-cell function (r ≥ 0.84 for SFA and r ≥ -0.71 for MUFA oleic ; P < 0.05) and insulin sensitivity (r ≥ -0.92 for SFA and r ≥ 0.89 for MUFA oleic ; P < 0.001) both in subjects with normal and high fasting triglycerides. In conclusion, dietary MUFA oleic , in contrast to SFA , favours the tuning towards better postprandial glycaemic control in subjects with normal and high fasting triglycerides.

Keyword: hyperlipedemia

Effect of triiodothyronine on triglyceride synthesis in human fibroblasts in different types of hypertriglyceridemia.

Fibroblasts from 12 normotriglyceridemic subjects and 30 hypertriglyceridemic patients and family members were used to investigate triglyceride synthesis and the influence of triiodothyronine on it. The monolayers were incubated for 72 hours with and without the thyroid hormone, followed by incorporation studies of radiolabeled acetic or into the cellular triglyceride fraction. Triiodothyronine had no influence on triglyceride synthesis of normal cell lines and of cells derived from patients with secondary hypertriglyceridemia, whereas fibroblasts from endogenous type IV patients showed higher rates of triglyceride synthesis under identical conditions. Values for type IV were in the range of 134% to 466% of the hormone-free control incubations. In cultures derived from patients with familial combined , no stimulation by triiodothyronine was observed: values were in the range of 64% to 144% of the hormone-free controls. Three out of four lines with type V gave "normal" values and are supposed to represent secondary hypertriglyceridemia, whereas one line may express endogenous type IV. The evidence obtained in vitro with cultured cells indicates different metabolic defects in endogenous type IV and familial combined ; it also shows the biochemically heterogenous nature of the disease "hypertriglyceridemia."

Keyword: hyperlipedemia

The rebound of lipoproteins after LDL-apheresis. Effects on chemical composition and LDL-oxidizability.

The changes in low density lipoprotein (LDL) composition and oxidizability after LDL-apheresis (LA) using dextran sulfate cellulose columns were evaluated in 12 hypercholesterolemic men (mean+/-S.D. total cholesterol (TC) 9.7+/-1.8 mmol/l). After 10-20 months on biweekly LA combined with simvastatin 40 mg per day immediate pre-apheresis levels of TC, LDL-cholesterol, and apolipoprotein B were decreased to 5.3+/-1.3 mmol/l, 3.3+/-1.2 mmol/l, and 1.6+/-0.4 g/l, respectively, whereas apheresis induced mean acute reductions of 61, 78, and 76%, respectively. Measurements of copper-induced LDL-oxidizability in vitro showed an increased resistance against oxidation after LA until day 3 post-treatment: lag time (min) (day 0 (before LA) versus day 1 (post-LA)) 112+/-27 versus 130+/-26 (P=0.001), maximal rate of diene production (nmol/min per mg LDL) 11.1+/-2.7 versus 9.1+/-2.1 (P=0.001), and time to maximal diene production (min) 186+/-39 versus 209+/-35 (P=0. 001). Analysis of the chemical composition of LDL revealed a 25% (P<0.001) reduced content of cholesteryl esters and a decrease of the cholesterol to protein ratio of 1.20+/-0.25 to 0.70+/-0.22 (P<0. 001) through the 3rd day post-LA. Linoleic and arachidonic content of LDL decreased 11 and 18%, respectively, at the expense of . Vitamin E levels (mg/l) were significantly lowered due to reduction of the lipoprotein pool by apheresis; however, vitamin E content of LDL did not change in the days after apheresis when expressed per g protein or per micromol linoleic . The changes in fatty pattern were strongly associated with changes in LDL-oxidizability indices (P

Keyword: hyperlipedemia

Fatty distribution in the major lipid classes in the hyperlipaemias.

Keyword: hyperlipedemia

[Lipoproteins and fatty acids--hyperlipoproteinemia and arteriosclerosis].

At first on the basis of literature a short description of the interrelations between lipoprotein metabolism and fatty metabolism as well as hyperlipoproteinaemia on the one hand and arteriosclerotic-degenerative diseases of the coronary vessels on the other is given. Finally, the findings of gas-chromatographic analyses of the patterns of fatty acids of the cholesterol ester and triglyceride fractions of the serum of 252 patients with primary hyperlipoproteinaemia are represented. In contrast to persons with healthy metabolism increase of the proportions per cent of , stearic , palmitoleic , oleic , and eikosatrienic was shown and decrease of those ones of linoleic , linolenic , arachidonic , and eikosapentaic in the cholesterol ester fractions in hyperlipoproteinaemia type IIa and IIb and in the triglyceride fractions in hyperlipoproteinaemia type IIb and IV. The shifts within the patterns of fatty acids in hyperlipoproteinaemia are an expression of an atherogenic risk. Disturbances of the hepatogenic synthesis of fatty acids and oxydation of fatty acids, increased reesterification from the depot fat of mobilised fatty acids as well as a partially increased LCAT-activity are causally discussed.

Keyword: hyperlipedemia

Beneficial effects of omega-3 fatty treatment on the recovery of cardiac function after cold storage of hyperlipidemic rats.

Cardiac effects of omega-3 polyunsaturated fatty acids (PUFAs) were studied in female Wister rats fed a standard diet (control [C] diet) or a high-cholesterol (HC) diet. Subgroups of rats from these groups were treated with eicosapentaenoic -E (EPA) or docosahexaenoic -95E (DHA) for 5 weeks. Although plasma total cholesterol (TC) and triglyceride (TG) levels were higher in each group fed the HC diet versus each group fed the C diet, EPA administration with the HC diet (HC + EPA) significantly (P < .05) reduced these levels. An isolated working-heart preparation was used to determine cardiac function. Cardiac output (CO) was lower in rats fed the HC diet and HC + DHA versus any of the groups fed the C diet (P < .05). In addition, left ventricular (LV) maximum differentiation of pressure-time curve (dp/dt) was lower in the rats fed the HC diet versus any of the C diet groups (P < .05). After evaluation of cardiac function in each rat, the heart was stored in a histidine-tryptophan-ketoglutarate solution for 8 hours at 4 degrees C. The heart was then reperfused, and recovery of cardiac function was evaluated. No significant differences were observed for post-preservative cardiac function within the C diet groups. However, within the HC diet groups, HC + EPA significantly (P < .05) improved the recovery of cardiac function. In addition, HC + DHA also significantly (P < .05) improved the recovery of coronary flow (CF) and LV dp/dt. No significant differences were observed for plasma TC and TG concentrations in the C diet groups. EPA administration significantly decreased cardiac levels of , oleic, and linoleic acids in the HC diet groups. No significant differences were observed for cardiac levels of free fatty acids (FFAs) within the C diet groups. Cardiac EPA and DHA levels were significantly (P < .05) elevated in EPA- or DHA-treated rats compared with the other diet-fed rats. Cardiac EPA levels were also elevated in DHA-treated rats compared with untreated rats (P < .05). These results suggest that EPA attenuates coronary and myocardial preservation injuries through an increase in serum lipids and an accumulation of myocardial FFAs resulting from a HC diet.

Keyword: hyperlipedemia

Modulation of postprandial lipaemia by a single meal containing a commonly consumed interesterified -rich fat blend compared to a non-interesterified equivalent.

Interesterification of palm stearin and palm kernal (PSt/PK) is widely used by the food industry to create fats with desirable functional characteristics for applications in spreads and bakery products, negating the need for trans fatty acids. Previous studies have reported reduced postprandial lipaemia, an independent risk factor for CVD, following interesterified (IE) and stearic -rich fats that are not currently widely used by the food industry. The current study investigates the effect of the most commonly consumed PSt/PK IE blend on postprandial lipaemia.A randomised, controlled, crossover (1\xa0week washout) double-blind design study (n\xa0=\xa012 healthy males, 18-45\xa0years), compared the postprandial (0-4\xa0h) effects of meals containing 50\xa0g fat [PSt/PK (80:20); IE vs. non-IE] on changes in plasma triacylglycerol (TAG), glucose, glucose-dependent insulinotropic polypeptide (GIP), peptide YY (PYY), insulin, gastric emptying (paracetamol concentrations) and satiety (visual analogue scales).The postprandial increase in plasma TAG was higher following the IE PSt/PK versus the non-IE PSt/PK, with a 51\xa0% greater incremental area under the curve [mean difference with 95\xa0% CI 41 (23, 58)\xa0mmol/L\xa0min P\xa0=\xa00.001]. The pattern of lipaemia was different between meals; at 4-h plasma TAG concentrations declined following the IE fat but continued to rise following the non-IE fat. Insulin, glucose, paracetamol, PYY and GIP concentrations increased significantly after the test meals (time effect; P\xa0<\xa00.001 for all), but did not differ between test meals. Feelings of fullness were higher following the non-IE PSt/PK meal (diet effect; P\xa0=\xa00.034). No other significant differences were noted.Interesterification of PSt/PK increases early phase postprandial lipaemia (0-4\xa0h); however, further investigation during the late postprandial phase (4-8\xa0h) is warranted to determine the rate of return to baseline values.Clinicaltrials.gov as .

Keyword: hyperlipedemia

[Lipolysis in very low density lipoproteins - locus minoris resistentiae - in the pathogenesis of hypertriglyceridemia. Positive effects of diet, polyenic fatty acids, statins and fibrates.]

Inhibition of hydrolysis of and oleic triglycedires (TG) in very low density lipoproteins (VLDL), slow formation of active apoВ-100 conformation, blockade of апоЕ/В-100 ligand formation in VLDL and their reduced uptake by insulin-dependent cells cause hypertriglyceridemia (HTG). and oleic VLDL (>80% total VLDL) are not converted in low density lipoproteins (LDL). Atherosclerosis is not an alimentary deficiency of polyenic fatty acids (PFA), but results from low in vivo bioavailability of PFA in LDL against the background of high dietary FA and LDL. Plasma PFA content and cellular PFA deficiency are as high as LDL cholesterol (CL). Primary prevention of atherosclerosis should be based on a decrease in dietary content of saturated FA, trans FA and a moderate increase in PFA. It seems highly unlikely that the xeobiotics statins, fibrates and probucol produce pleiotropic biological effects in vivo. These effects are brought about by phylogenetically early humoral mediators eicosanoids: prostacyclins, prostaglandins, thromboxanes, leukotrienes, and resolvins. It is reasonable to suggest that all preparations which act according to the same algorithm activate TG hydrolysis in VLDL and normalize cellular uptake of PFA in linoleic and linolenic LDL via apoВ-100 endocytosis. Atherosclerosis is a syndrome of cellular deficiency of essential polyenic FA.

Keyword: hyperlipedemia

The chain length of dietary saturated fatty acids affects human postprandial lipemia.

Saturated fats increase total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C) and are linked to coronary artery disease risk. The effect of variance in chain length of saturated fatty acids (SFA) on coronary artery disease in human postprandial lipemia is not well elucidated.A total of 20 healthy volunteers were challenged with 3 test meals, similar in fat content (~31% en) but varying in saturated SFA content and polyunsaturated/saturated fatty ratios (P/S). The 3 meals were lauric + myristic -rich (LM), P/S 0.19; -rich (POL), P/S 0.31; and stearic -rich (STE), P/S 0.22. Blood was sampled at fasted baseline and 2, 4, 5, 6, and 8 hours. Plasma lipids (triacylglycerol [TAG]) and lipoproteins (TC, LDL-C, high density lipoprotein-cholesterol [HDL-C]) were evaluated.Varying SFA in the test meal significantly impacted postprandial TAG response (p < 0.05). Plasma TAG peaked at 5 hours for STE, 4 hours for POL, and 2 hours for LM test meals. Area-under-the-curve (AUC) for plasma TAG was increased significantly after STE treatment (STE > LM by 32.2%, p = 0.003; STE > POL by 27.9%, p = 0.023) but was not significantly different between POL and LM (POL > LM by 6.0%, p > 0.05). At 2 hours, plasma HDL-C increased significantly after the LM and POL test meals compared with STE (p < 0.05). In comparison to the STE test meal, HDL-C AUC was elevated 14.0% (p = 0.005) and 7.6% (p = 0.023) by the LM and POL test meals, respectively. The TC response was also increased significantly by LM compared with both POL and STE test meals (p < 0.05).Chain length of saturates clearly mediated postmeal plasma TAG and HDL-C changes.

Keyword: hyperlipedemia

Propylene glycol guluronate sulfate (PGGS) reduces lipid accumulation via AMP-activated kinase activation in palmitate-induced HepG2 cells.

Cardiovascular disease (CVD) is the No. 1 cause of death worldwide. is one of the major risk factors for CVD. Maintaining lipid homeostasis is an effective way to prevent CVD. We prepared propylene glycol guluronate sulfate (PGGS), a sulfated polysaccharide, and investigated its effect on lipid metabolism in HepG2 cells. We found that total cholesterol (TC) and triglycerides (TG) were significantly decreased in the cells after PGGS treatment. We have also shown that the AMPK signaling is activated after PGGS treatment as evidenced by changes in the expression of many AMPK downstream targets including SREBP-1c, SIRT-1, CPT1, PPARα, and FAS. Our results have demonstrated that PGGS is a potentially novel lipid-lowering agent for CVD prevention.Copyright © 2018. Published by Elsevier B.V.

Keyword: hyperlipedemia

Cholesteryl ester and triacylglycerol fatty acids in type V .

The fatty compositions of plasma cholesteryl esters (CF) and triacylglycerols (TG) from seven healthy individuals and five patients with type V hyperlipoproteinemia were determined. Very low density (VLDL), low density (LDL) and high density lipoproteins (HDL) wer isolated. The lipids were extracted from the lipoproteins and the triacylglycerols and cholesteryl esters separated for analysis. The fatty compositions of triacylglycerols from healthy and type V individuals were very similar. The cholesteryl esters from type V patients had increased contents of and decreased amounts of linoleic and arachidonic acids as compared to the normal individuals. The fatty composition of the cholesteryl esters from high density lipoproteins had the greatest deviation. The fatty compositions of the triacylglycerols from the two groups were similar. However, the triacylglycerols in all lipoprotein fractions contained more and oleic and less linoleic and arachidonic acids than the cholesteryl esters.

Keyword: hyperlipedemia

Palm oil and : a review on cardiovascular effects and carcinogenicity.

We reviewed the scientific literature on the evidence of the relationship between palm oil and adverse effects on human health. Few studies have investigated the effects of palm oil per se, and the main reason why it has been associated with negative health effects is the relatively high content of saturated fatty acids (SFAs), particularly , which in turn have been associated with increased risk of coronary heart disease and some tumours. However, more recent investigations on the topic seem to have reconsidered the negative role of the dietary SFAs as a risk factor for cardiovascular diseases and show that not only the type of fat, but also that the triglyceride structure plays a role in cholesterolaemia. As regards to a role in cancer, specific studies on dietary or palm oil and the risk of cancer development are scanty, and the evidence is not convincing.

Keyword: hyperlipedemia

Medium-chain dicarboxylic acylcarnitines as markers of n-3 PUFA-induced peroxisomal oxidation of fatty acids.

Omega-3 polyunsaturated fatty acids (n-3 PUFA) found in fish oil activate PPAR-α, stimulate peroxisomal fatty (FA) β-oxidation and prevent impairments on glucose homeostasis.Glucose metabolism and FA oxidation were studied in C57/Bl6 mice fed with diets containing either 3.6 and 31.5% fish oil or lard. To assess the effects of peroxisomal proliferation on FA oxidation independent of n-3 PUFA intake, mice were treated with the PPAR-α agonist WY-14643. n-3 PUFA-fed mice were protected from glucose intolerance and dyslipidemia compared to animals fed a lard-based high-fat diet. Most importantly, mice fed on the hyperlipidic diet based on fish oil as well as the WY-14643 treated mice showed twofold increase of odd, medium-chain, dicarboxylic acylcarnitines in the liver suggesting that not only β-oxidation, but also α- and ω-oxidation of FA were increased. Finally, an oxidation assay using liver homogenates and as substrate revealed an over tenfold increased production of similar acylcarnitines, indicating that FA are their precursors.This study shows at the metabolite level that peroxisome proliferation induced either by fish oil or WY-14643 is associated with increased α- and ω-oxidation of FA producing specific acylcarnitines that can be utilized as biomarkers of peroxisomal FA oxidation.© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: hyperlipedemia

Nutrition in lipid disorders.

When various fats and oils are given to humans, marked differences in the concentrations of serum lipids and lipoproteins will be observed. Relative to dietary carbohydrates, lauric, myristic and , which are saturated fatty acids, but also trans fatty acids from hydrogenated fats elevate the serum total and LDL cholesterol level. Saturated fatty acids with less than 12 carbon atoms or with 18 carbon atoms-stearic -have little effect on the serum cholesterol level. This is also true for oleic , a monounsaturated fatty . Replacement of linoleic for carbohydrates leads to slightly lower LDL cholesterol levels; however, recent studies suggest that this effect is less than predicted by studies performed in the early sixties. Substitution of carbohydrates by fat will increase the level of HDL cholesterol but decrease that of fasting serum triglyceride. Trans fatty acids, though, are an exception and lower the level of HDL cholesterol relative to other fatty acids. The fish oils may have a small LDL-cholesterol-elevating effect in hypertriglyceridemic patients but have a very specific triglyceride-lowering effect.

Keyword: hyperlipedemia

Effects of increasing dietary palmitoleic compared with and oleic acids on plasma lipids of hypercholesterolemic men.

Palmitoleic is a minor monounsaturated fatty in the human diet and in blood plasma. Because macadamia oil is at least one potentially large source of palmitoleic , we tested its effect on plasma lipid levels against two other dietary fatty acids, oleic and . The dietary adjustments, through the use of supplements, provided comparisons of the three test fatty acids in which palmitoleic could be judged as behaving either like a saturated or a monounsaturated . Thirty-four hypercholesterolemic men ate the three test diets in random order in 3-week periods. Plasma total cholesterol and low density lipoprotein (LDL) cholesterol concentrations were similar with and palmitoleic acids and significantly higher than with oleic . High density lipoprotein (HDL) cholesterol was significantly lower with palmitoleic than with . The study confirms that, at least in hypercholesterolemic men, a modest increase in (+4% en) raises LDL cholesterol relative to oleic (+3% en), even when dietary cholesterol is low (< 165 mg/day). Palmitoleic (+4% en) behaves like a saturated and not a monounsaturated fatty in its effect on LDL cholesterol.

Keyword: hyperlipedemia

The fatty pattern of triglycerides in liver, adipose tissue and serum of diabetics with hyperlipoproteinemia before and during clofibrate treatment.

In 20 diabetic inpatients with type IIb, III, IV and V hyperlipoproteinemia (HPL) the triglyceride fatty pattern (TFAP) of serum, adipose tissue and liver biopsy specimens before and after one year of clofibrate treatment has been determined by gas-liquid chromatography. Compared to previous results which revealed a correlation between fat droplet size and the TFAP in liver parenchyma cells, remarkable changes were observed after long-term therapy. In adipose tissue, only linoleic increased significantly from 8.6 to 11.3%. In serum, myristic and decreased, whereas linoleic, eicosatetraenoic (arachidonic) and eicosapentaenoic rose significantly. In liver fat, decreased, whereas linoleic, eicosatrienoic, arachidonic and eicosapentaenoic significantly increased. After clofibrate therapy, the TFAP in diabetic subjects with HLP became similar to that of diabetics without HLP. The most pronounced changes were found in the liver, serum having an intermediate position between liver and adipose tissue. The pathophysiological relevance in view of possible relations to prostaglandins has been discussed.

Keyword: hyperlipedemia

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, , and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: hyperlipedemia

Effect of dietary Fatty acids on human lipoprotein metabolism: a comprehensive update.

Dyslipidemia is a major risk factor for cardiovascular disease (CVD). Dietary fatty- composition regulates lipids and lipoprotein metabolism and may confer CVD benefit. This review updates understanding of the effect of dietary fatty-acids on human lipoprotein metabolism. In elderly participants with , high n-3 polyunsaturated fatty-acids (PUFA) consumption diminished hepatic triglyceride-rich lipoprotein (TRL) secretion and enhanced TRL to low-density lipoprotein (LDL) conversion. n-3 PUFA also decreased TRL-apoB-48 concentration by decreasing TRL-apoB-48 secretion. High n-6 PUFA intake decreased very low-density lipoprotein (VLDL) cholesterol and triglyceride concentrations by up-regulating VLDL lipolysis and uptake. In a study of healthy subjects, the intake of saturated fatty-acids with increased at the sn-2 position was associated with decreased postprandial lipemia. Low medium-chain triglyceride may not appreciably alter TRL metabolism. Replacing carbohydrate with monounsaturated fatty-acids increased TRL catabolism. Trans-fatty- decreased LDL and enhanced high-density lipoprotein catabolism. Interactions between APOE genotype and n-3 PUFA in regulating lipid responses were also described. The major advances in understanding the effect of dietary fatty-acids on lipoprotein metabolism has centered on n-3 PUFA. This knowledge emphasizes the importance of regulating lipoprotein metabolism as a mode to improve plasma lipids and potentially CVD risk. Additional studies are required to better characterize the cardiometabolic effects of other dietary fatty-acids.

Keyword: hyperlipedemia

Liraglutide ameliorates palmitate-induced endothelial dysfunction through activating AMPK and reversing leptin resistance.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue, is an antidiabetic drug. It has been shown to improve endothelial dysfunction, but the mechanism remains somewhat unclear. Leptin can also improve endothelial function. Cardiovascular disease (CVD) is linked to hyperleptinemia, and leptin resistance, how liraglutide influences the effect of leptin on endothelial function, is never reported. We used (PA) to mimic in endothelial cells to explore the cardio-protective mechanism of liraglutide and its impact on the role of leptin.Human umbilical vein endothelial cells (HUVECs) were incubated with PA for 16\xa0h and then were treated with liraglutide for 30\xa0min.PA elevated not only phosphorylation of JNK and IKKα/β, but also the expression of IL-6 in HUVECs. These effects of PA were reversed by liraglutide. In addition, liraglutide increased phosphorylation of eNOS, AMPK, and the release of NO but had no effect on PKC phosphorylation. In addition, leptin elevated eNOS phosphorylation but was abrogated by PA. However, in the presence of liraglutide, leptin regained its function of elevating eNOS phosphorylation. Last, we found that liraglutide inhibited PA-elevated SOCS3, which is a marker of leptin resistance.GLP-1 impairs endothelial inflammatory signals, improves endothelial function, and reverses leptin resistance.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Effect of alipogene tiparvovec (AAV1-LPL(S447X)) on postprandial chylomicron metabolism in lipoprotein lipase-deficient patients.

Lipoprotein lipase-deficient (LPLD) individuals display marked chylomicronemia and hypertriglyceridemia associated with increased pancreatitis risk. The aim of this study was to determine the effect of i.m. administration of an adeno-associated viral vector (AAV1) for expression of LPL(S447X) in muscle (alipogene tiparvovec, AAV1-LPL(S447X)) on postprandial chylomicron metabolism and on nonesterified fatty (NEFA) and glycerol metabolism in LPLD individuals.In an open-label clinical trial (CT-AMT-011-02), LPLD subjects were administered alipogene tiparvovec at a dose of 1 × 10(12) genome copies per kilogram. Two weeks before and 14 wk after administration, chylomicron metabolism and plasma palmitate and glycerol appearance rates were determined after ingestion of a low-fat meal containing (3)H-palmitate, combined with (continuous) iv infusion of [U-(13)C]palmitate and [1,1,2,3,3-(2)H]glycerol.After administration of alipogene tiparvovec, the triglyceride (TG) content of the chylomicron fraction and the chylomicron-TG/total plasma TG ratio were reduced throughout the postprandial period. The postprandial peak chylomicron (3)H level and chylomicron (3)H area under the curve were greatly reduced (by 79 and 93%, 6 and 24 h after the test meal, respectively). There were no significant changes in plasma NEFA and glycerol appearance rates. Plasma glucose, insulin, and C-peptide also did not change.Intramuscular administration of alipogene tiparvovec resulted in a significant improvement of postprandial chylomicron metabolism in LPLD patients, without inducing large postprandial NEFA spillover.ClinicalTrials.gov .

Keyword: hyperlipedemia

Mitochondrial dysfunction in insulin resistance: differential contributions of chronic insulin and saturated fatty exposure in muscle cells.

Mitochondrial dysfunction has been associated with insulin resistance, obesity and diabetes. Hyperinsulinaemia and are hallmarks of the insulin-resistant state. We sought to determine the contributions of high insulin and saturated fatty exposure to mitochondrial function and biogenesis in cultured myocytes. Differentiated C2C12 myotubes were left untreated or exposed to chronic high insulin or high palmitate. Mitochondrial function was determined assessing: oxygen consumption, mitochondrial membrane potential, ATP content and ROS (reactive oxygen species) production. We also determined the expression of several mitochondrial genes. Chronic insulin treatment of myotubes caused insulin resistance with reduced PI3K (phosphoinositide 3-kinase) and ERK (extracellular-signal-regulated kinase) signalling. Insulin treatment increased oxygen consumption but reduced mitochondrial membrane potential and ROS production. ATP cellular levels were maintained through an increased glycolytic rate. The expression of mitochondrial OXPHOS (oxidative phosphorylation) subunits or Mfn-2 (mitofusin 2) were not significantly altered in comparison with untreated cells, whereas expression of PGC-1α (peroxisome-proliferator-activated receptor γ co-activator-1α) and UCPs (uncoupling proteins) were reduced. In contrast, saturated fatty exposure caused insulin resistance, reducing PI3K (phosphoinositide 3-kinase) and ERK (extracellular-signal-regulated kinase) activation while increasing activation of stress kinases JNK (c-Jun N-terminal kinase) and p38. Fatty acids reduced oxygen consumption and mitochondrial membrane potential while up-regulating the expression of mitochondrial ETC (electron chain complex) protein subunits and UCP proteins. Mfn-2 expression was not modified by palmitate. Palmitate-treated cells also showed a reduced glycolytic rate. Taken together, our findings indicate that chronic insulin and fatty -induced insulin resistance differentially affect mitochondrial function. In both conditions, cells were able to maintain ATP levels despite the loss of membrane potential; however, different protein expression suggests different adaptation mechanisms.

Keyword: hyperlipedemia

Fetuin A promotes lipotoxicity in β cells through the TLR4 signaling pathway and the role of pioglitazone in anti-lipotoxicity.

Fetuin A (FetA), a secreted glycoprotein, is known to affect inflammation and insulin resistance (IR) in obese humans and animals. Lipotoxicity from chronic damages pancreatic β cells, hastening the onset of diabetes. We sought to determine whether FetA promotes lipotoxicity through modulation of the toll-like receptor 4 (TLR4) inflammatory signaling pathway as well as the protective effect of pioglitazone(PIO) on lipotoxicity.βTC6, a glucose-sensitive mouse pancreatic β cell line, and Sprague-Dawley rats with diet-induced obesity, were used to investigate FetA-mediated lipotoxicity. Protein expression/activation were measured by Western blotting. Small interfering (si)RNAs for TLR4 were used. Cell apoptosis was quantified by TUNEL analysis or flow cytometry, respectively. Insulin release was assessed with an insulin ELISA.FetA dose-dependently aggravated (PA)-induced βTC6 cell apoptosis, insulin secretion impairment, and inhibition of the expression of G-protein-coupled receptor 40 (GPR40) and pancreatic duodenal homeobox-1(PDX-1). Combined FetA\u2009+\u2009PA induced TLR4 expression, and subsequent inhibition of TLR4 signaling or expression was shown to prevent the strengthening effect of FetA on PA-induced lipotoxicity in βTC6 cells. FetA\u2009+\u2009PA induced p-JNK and nuclear factor-κB (NF-κB) subunit P65 expression, and inhibition of this activity reduced PA+ FetA lipotoxicity in βTC6 cells. PIO could ameliorate PA+ FetA-induced damage to βTC6 cells. Similarly, PIO improved insulin secretion disorder, reduced apoptosis, decreased FetA, TLR4, p-JNK, NF-κB subunit P65 and cleaved caspase 3 expression, and increased GPR40 and PDX-1 expression in islet β cells of diet-induced obese rats. The correlative bivariate analysis showed that increases in Fetuin A were directly proportional to the development of β cell injury.FetA can promote lipotoxicity in β cells through the TLR4-JNK-NF-κB signaling pathway. The protective effects of PIO on lipotoxicity in β cells may involve the inhibition of the activation of the FetA and TLR4 signaling pathway.Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Keyword: hyperlipedemia

Effects of 2,3-dihydrophthalazine-1,4-dione on Sprague Dawley rats lipid metabolism and serum lipoproteins.

2,3-Dihydrophthalazine-1,4-dione effectively lowers serum levels of cholesterol and triglycerides in Sprague Dawley rats after two weeks, after which the cholesterol levels continued to decline. The maximum serum lipid lowering effect on cholesterol or triglyceride levels was during the seventh and eighth week of drug administration. Similar magnitudes of reduction in lipids were observed in hyperlipidemic diet induced mice after four weeks of drug administration of 20 mg/kg.day. Lipid levels in liver, small intestines and aorta wall tissue were significantly reduced after eight weeks of drug administration, but no significant increase in fecal lipids was noted. The cholesterol content in the chylomicrons and in the VLDL and LDL-fractions was significantly reduced whereas HDL cholesterol was elevated by 112%. Neutral lipids and the triglyceride content were not altered in the chylomicron, VLDL and LDL; however, reductions of both levels were observed in the HDL fraction. The phospholipid content was reduced in the LDL and elevated in the other three fractions. Incorporation studies into lipoprotein fractions showed a decrease in cholesterol incorporation in chylomicrons, VLDL and LDL with an increase in HDL cholesterol incorporation. incorporation was reduced in the chylomicron, VLDL and HDL fractions. 32P-Incorporation was reduced in the HDL fraction. Leucine incorporation into the apoproteins of all four fractions was elevated. Rate limiting enzymes involved in de novo cholesterol, fatty and triglyceride synthesis were inhibited by 2,3-dihydrophthalazine-1,4-dione after eight weeks of administration. There was no evidence that the drug caused an increase in peroxisome formation as measured by liver catalase activity nor the release of lysosomal hydrolytic enzymes as measured by phosphatase and cathepsin activities. The drug afforded no deleterious effects on clinical chemistry parameters after eight weeks administration.

Keyword: hyperlipedemia

Application of GC/MS-based metabonomic profiling in studying the lipid-regulating effects of Ginkgo biloba extract on diet-induced in rats.

To evaluate the lipid-regulating effects of extract from Ginkgo biloba leaves (EGB) using pharmacological methods and metabonomic profiling in a rat model of diet-induced .EGB was orally administered at a dose level of 40 mg/kg in both the EGB-prevention and -treatment groups. All rat samples obtained were examined for known and potential biomarkers and enzyme activity using commercial assay kits and GC/MS-based metabonomic profiling coupled with principal component analysis (PCA).The data obtained from the assay kits indicated that EGB reduced total cholesterol and low density lipoprotein cholesterol levels and increased high density lipoprotein cholesterol levels in rat plasma obtained from both the EGB-prevention and -treatment groups compared with those of the diet-induced group. EGB also increased the activities of lipoprotein lipase and hepatic lipase and excretion of fecal bile in rats from the EGB-prevention and-treatment groups. Using GC/MS-based metabonomic analysis, more than 40 endogenous metabolites were identified in rat plasma. PCA of rat plasma samples obtained using GC/MS produced a distinctive separation of the four treatment groups and sampling points within each group. Metabolic changes during formation and improvement resulting from EGB treatment were definitively monitored with PCA score plots. Furthermore, elevated levels of sorbitol, tyrosine, glutamine and glucose, and decreased levels of citric , galactose, , arachidonic , acetic , cholesterol, butyrate, creatinine, linoleate, ornithine and proline, were observed in the plasma of rats treated with EGB.EGB exerts multi-directional lipid-lowering effects on the rat metabonome, including limitation of the absorption of cholesterol, inactivation of HMGCoA and favorable regulation of profiles of essential polyunsaturated fatty (EFA). Further experiments are warranted to explore the mechanisms of action underlying the lipid-regulating effects of EGB against .

Keyword: hyperlipedemia

Acute hypoxic preconditioning prevents -induced cardiomyocyte apoptosis via switching metabolic GLUT4-glucose pathway back to CD36-fatty dependent.

Metabolic syndrome is a risk factor for the development of cardiovascular diseases. Myocardial cell damage leads to an imbalance of energy metabolism, and many studies have indicated that short-term hypoxia during myocardial cell injury has a protective effect. In our previous animal studies, we found that short-term hypoxia in the heart has a protective effect, but long-term hypoxia increases myocardial cell injury. (PA)-treated H9c2 cardiomyoblasts and neonatal rat ventricle cardiomyocytes were used to simulate model, which suppress cluster of differentiation 36 (CD36) and activate glucose transporter type 4 (GLUT4). We exposed the cells to short- and long-term hypoxia and investigated the protective effects of hypoxic preconditioning on PA-induced lipotoxicity in H9c2 cardiomyoblasts and neonatal rat cardiomyocytes. Preconditioning with short-term hypoxia enhanced both CD36 and GLUT4 metabolism pathway protein levels. Expression levels of phospho-PI3K, phospho-Akt, phospho-AMPK, SIRT1, PGC1α, PPARα, CD36, and CPT1β induced by PA was reversed by short-term hypoxia in a time-dependent manner. PA-induced increased GLUT4 membrane protein level was reduced in the cells exposed to short-term hypoxia and si-PKCζ. Short-term hypoxia, resveratrol and si-PKCζ rescue H9c2 cells from apoptosis induced by PA and switch the metabolic pathway from GLUT4 dependent to CD36 dependent. We demonstrate short-term hypoxic preconditioning as a more efficient way as resveratrol in maintaining the energy metabolism of hearts during and can be used as a therapeutic strategy.© 2017 Wiley Periodicals, Inc.

Keyword: hyperlipedemia

Potential for enhancing the nutritional properties of milk fat.

Milk fat has been identified as a hypercholesterolemic fat because it contains cholesterol and is primarily saturated. However, different types of dietary saturated fats do not have equivalent effects on plasma cholesterol levels relevant to ingestion of polyunsaturated fats. Research suggests that the hypercholesterolemic effect of saturated fats in human diets is largely due to 12, 14, and 16 carbon chain length fatty acids. Evidence also suggests that stearic (C18:0) is as effective as oleic (C18:ln-9) in lowering plasma cholesterol levels when either replaces (C16:0) in the diet of men. Milk fat has a unique fatty profile with approximately 10% short- and medium-chain length saturated fatty acids (less than 12 carbons) and 35% of total fatty acids from stearic and oleic acids. The contribution of milk products to fat and cholesterol intake in the typical American diet is less than that provided by other animal products. This paper will review the recommendations of the National Cholesterol Education Program, the effects of milk fat ingestion on blood cholesterol, and the rationale and feasibility of three approaches to modifying the lipid composition of milk fat.

Keyword: hyperlipedemia

Therapeutic effects of quercetin on early inflammation in hypertriglyceridemia-related acute pancreatitis and its mechanism.

To investigate the therapeutic effects of quercetin on early-stage inflammation in hypertriglyceridemia (HTG)-related acute pancreatitis (AP) both in\xa0vivo and in\xa0vitro, and its possible mechanism.In\xa0vivo, rats were fed a high-fat diet to induce HTG, and AP was induced by intraperitoneal injection of cerulein (50\xa0μg/kg\xa0×\xa02). Quercetin (100, 150 and 200\xa0mg/kg) was administered by intraperitoneal injection after AP induction. In\xa0vitro, rat exocrine acinar cells were preincubated with (PA, 0.1\xa0mmol/L, 6\xa0h) with quercetin (5, 10, 20 and 40\xa0μM) prior to a cholecystokinin analog CCK-8 (20pM). Injury of the pancreas was assessed by amylase secretion and pancreatic histological evaluation. Inflammation was estimated by measuring IL-1β, IL-6, TNFα and NF-kB expression. Dynamic expression of IRE1α, sXBP1, C/EBPα and C/EBPβ was monitored by real-time PCR, immunofluorescence (IF) and western blot (WB).Quercetin intervention reduced plasma amylase level (P\xa0<\xa00.001) in a dose-dependent manner, attenuated pancreatic histopathological damage (P\xa0<\xa00.05), and reduced the mRNA and protein expression of NF-kB, IL-1β, IL-6, TNFα (P\xa0<\xa00.05) more significantly in HTG-related AP rats than in normal-lipid AP rats. Quercetin also down-regulated gene and protein expression levels of IRE1α, sXBP1, C/EBPα and C/EBPβ in a dose-dependent manner.Quercetin attenuates early-stage inflammation in HTG-related AP, probably by reducing IRE1α, sXBP1, C/EBPα and C/EBPβ expression.Copyright © 2016 IAP and EPC. Published by Elsevier India Pvt Ltd. All rights reserved.

Keyword: hyperlipedemia

Genipin ameliorates age-related insulin resistance through inhibiting hepatic oxidative stress and mitochondrial dysfunction.

Insulin resistance (IR) increases with age and plays a key role in the pathogenesis of type 2 diabetes mellitus. Oxidative stress and mitochondrial dysfunction are supposed to be major factors leading to age-related IR. Genipin, an extract from Gardenia jasminoides Ellis fruit, has been reported to stimulate insulin secretion in pancreatic islet cells by regulating mitochondrial function. In this study, we first investigated the effects of genipin on insulin sensitivity and the potential mitochondrial mechanisms in the liver of aging rats. The rats were randomly assigned to receive intraperitoneal injections of either 25mg/kg genipin or vehicle once daily for 12days. The aging rats showed hyperinsulinemia and , and insulin resistance as examined by the decreased glucose decay constant rate during insulin tolerance test (kITT). The hepatic tissues showed steatosis and reduced glycogen content. Hepatic malondialdehyde level and mitochondrial reactive oxygen species (ROS) were higher, and levels of mitochondrial membrane potential (MMP) and ATP were lower as compared with the normal control rats. Administration of genipin ameliorated systemic and hepatic insulin resistance, alleviated hyperinsulinemia, hyperglyceridemia and hepatic steatosis, relieved hepatic oxidative stress and mitochondrial dysfunction in aging rats. Furthermore, genipin not only improved insulin sensitivity by promoting insulin-stimulated glucose consumption and glycogen synthesis, inhibited cellular ROS overproduction and alleviated the reduction of levels of MMP and ATP, but also reversed oxidative stress-associated JNK hyperactivation and reduced Akt phosphorylation in palmitate-treated L02 hepatocytes. In conclusion, genipin ameliorates age-related insulin resistance through inhibiting hepatic oxidative stress and mitochondrial dysfunction.© 2013.

Keyword: hyperlipedemia

Plasma cholesterol-lowering potential of edible-oil blends suitable for commercial use.

We tested semihardened blends of edible oils, suitable for commercial food manufacture, with a lower-than-conventional saturated fatty content, for their effects on plasma cholesterol. Twenty-six mildly hypercholesterolemic men took part in a double-blind crossover experiment in which two test blends were compared with two control dietary periods [which resembled the Australian fat intake: proportions of polyunsaturated, monounsaturated, and saturated fatty acids (PMS) 0.4:0.9:1]. PMS in the test diets was approximately 0.8:1.3:1 and resulted in significantly lower LDL-cholesterol concentrations (reductions of less than or equal to 7.7%). HDL cholesterol and plasma triglyceride were unchanged. The trans fatty (mainly elaidic) content of the blends was 16%, raising its contribution to energy by 4% but without apparent effect on LDL and HDL concentrations. Provided the overall ratio of linoleic to in commercial edible-oil blends exceeds that in the prevailing national diet, partial hydrogenation will not negate the LDL-lowering potential.

Keyword: hyperlipedemia

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, , and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, , hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: hyperlipedemia

Mechanism of avian estrogen-induced hypertriglyceridemia: evidence for overproduction of triglyceride.

Relying on methods other than the determination of turnover rate of triglyceride from the curve of plasma triglyceride radioactivity after administration of labeled precursor, we have confirmed that the endogenous hypertriglyceridemia induced by estrogenization of the chick is accompanied by increased production of triglyceride. Chicks estrogenized with diethylstilbestrol became grossly hypertriglyceridemic and had elevated levels of plasma free fatty . Within 5 min of administration of labeled palmitate, estrogenized hypertriglyceridemic birds converted approximately 10 times more plasma free fatty to hepatic triglyceride than did controls. In addition, 2 hr after intraperitoneal injection of [14-C]acetate or [U-14-C]glucose, the specific activity of very low density lipoprotein triglyceride (VLDL-TG) of estrogenized birds reached or exceeded that of the untreated controls, and the rapid enrichment of the vastly expanded plasma VLDL-TG pool with labeled triglyceride further indicated that increased production of triglyceride occurs with estrogenization. Furthermore, [14-C]acetate incorporation into VLDL-TG was calculated to be 1.6 and 6.6% of the injected dose in estrogenized birds compared with 0.1 and 0.2% in untreated birds. Increased production of plasma VLDL-TG was confirmed by a kinetic study of VLDL-TG metabolism, employing reinjected, endogenously prepared [14-C]triglyceride-labeled VLDL. The fractional turnover rate of VLDL-TG in estrogenized hypertriglyceridemic birds was substantially less than that in untreated controls (0.32 plus or minus 0.03 vs 0.71 plus or minus 0.03/hr), but the total turnover rate was nearly 50 times greater (244 plus or minus 52 vs. 5 plus or minus 1 mg/hr).

Keyword: hyperlipedemia

Novel hypolipidemic conjugates of fatty and bile with lysine for linkage.

Novel fatty -bile conjugates (1a-1k) were designed and synthesized by coupling of the fatty acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve liver-specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high-fat diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced liver damage.

Keyword: hyperlipedemia

[Changes in the relative levels of fatty acids in blood and myocardium in the Prague breed of rats with hereditary hypercholesteremia after administration of slow calcium channel blockers].

The present paper describes the effect of six-week oral administration of verapamil and diltiazem (1 mg.kg-1 of weight two times daily in 12 hour intervals) on the content of fatty acids of the serum and myocardium of PHHC rats. A cholesterol diet changes the content of fatty acids of the serum and myocardium of PHHC rats in comparison with control rats without the cholesterol diet. A significant decrease in the content of , a decrease in the content of stearic , linoleic and arachidonic and a significant increase in the content of oleic were observed in the serum. Long-term administration of the slow calcium channel blockers produces another decrease in the content of the bound form of arachidonic . Changes in the representation of other fatty acids are not marked. Long-term administration of a cholesterol diet produces an increase in the content of and stearic and a decrease in the content of oleic , linoleic and arachidonic in the myocardium. Administration of verapamil results in a modification of the above-mentioned changes in all parameters excepting the content of arachidonic , the content of which was decreased in an even more marked manner. Administration of diltiazem produced an accumulation of both saturated and mono-unsaturated fatty acids (, stearic and oleic acids) and produced a significant decrease in the content of linoleic and mainly the bound form of arachidonic .(ABSTRACT TRUNCATED AT 250 WORDS).

Keyword: hyperlipedemia

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism.A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and , accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: hyperlipedemia

miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 diabetes mellitus through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 diabetes mellitus (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid metabolism were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of lipid droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and lipid levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic lipid droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid metabolism dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid metabolism in T2DM, which can inhibit hepatic lipogenesis and gluconeogenesis and elevate glycogen synthesis by targeting STAT3.

Keyword: hyperlipedemia

P38 plays an important role in glucolipotoxicity-induced apoptosis in INS-1 cells.

The mechanism underlying the regulation of glucolipotoxicity-induced apoptosis by MAPKs was examined in INS-1 cells.The rat insulinoma cell line INS-1 was cotreated with glucose (30 mM) and (0.2 mM) (GLU+PA). Apoptosis was assessed by cell morphology and detection of PARP cleavage. The activation of MAPKs was examined by Western blotting using specific antibodies against the phosphorylated forms of JNK, ERK1/2, and P38.(1) Live cell imaging studies showed that treatment with GLU+PA for 72 h induced significant cell death, concomitant with PARP-1 cleavage and caspase-3 activation, which peaked at 96 h of treatment. (2) Western blot analysis of the activation of MAPKs during GLU+PA-induced INS-1 cell apoptosis showed that phosphorylation of P38 increased gradually and reached a peak at 96 h, which coincided with PARP-1 cleavage. A transient increase of ERK activation was followed by a rapid decline at 96 h, whereas JNK phosphorylation status remained unchanged in response to GLU+PA. (3) Phosphorylation of insulin receptor substrate (IRS)-2 at 48 h of treatment triggered its degradation, which coincided with P38 activation. (4) Inhibition of P38, but not JNK or ERK, blocked GLU+PA-induced INS-1 cell apoptosis.P38 may be involved in the regulation of glucolipotoxicity-induced apoptosis through the phosphorylation of IRS-2.

Keyword: hyperlipedemia

[Effect of clofibric on fatty metabolism in HLP patients in the 2d half of life].

Ten patients with primary type IIb hyperlipoproteinemia and one hundred patients with primary type IV hyperlipoproteinemia (sixty-seven men and forty-three women aged forty-three to seventy-four) were treated with clofibric (Regadrin) for three years. Gas chromatographic analysis of the composition of cholesterol ester and triglyceride fatty acids in serum were done prior to and at four-month intervals during therapy. During treatment of type IIb and type IV hyperlipoproteinemia there was observed a decrease of , stearic, palmitoleic, oleic, and eicosane-tetraenoic acids as well as an increase of linoleic, linolenic, arachidonic, and eicosane-pentaenoic acids in the triglyceride fraction. These changes manifest themselves most conspicuously in the effect of clofibric upon the hepatogenic fatty metabolism (increased synthesis or reduced catabolization of polyunsaturated fatty acids and increased oxidation or reduced formation of monounsaturated and saturated fatty acids, increased esterification of polyenic acids). In addition, these is the possibility of selective displacement of saturated and monounsaturated fatty acids of the unsaturated fatty acids fraction as well as specific inhibition of their esterification with glycerol by clofibric . Treatment with clofibric over a long period of time may well give rise to additional reactions through influences exerted upon the insulin level.

Keyword: hyperlipedemia

Differential effects of saturated fatty acids on low density lipoprotein metabolism in the guinea pig.

Studies have shown that dietary fat saturation affects guinea pig plasma low density lipoprotein (LDL) levels by altering both LDL receptor-mediated catabolism and flux rates of LDL (Fernandez et al. 1992. J. Lipid Res. 33: 97-109). The present studies investigated whether saturated fatty acids of varying chain lengths have differential effects on LDL metabolism. Guinea pigs were fed 15% (w/w, 35% calories) fat diets containing either palm kernel oil (PK), 52% lauric /18% myristic ; palm oil (PO), 43% /4% stearic ; or beef tallow (BT), 23% /14% stearic . Plasma LDL cholesterol levels were significantly higher for animals fed the PK diet (P < 0.001) with values of 83 +/- 19 (n = 12), 53 +/- 8 (n = 12) and 44 +/- 16 (n = 10) mg/dl for PK, PO, and BT diets, respectively. The relative percentage composition of LDL was modified by fat type; however, LDL diameters and peak densities were not different between diets, indicating no effect of saturated fatty composition on LDL size. ApoB/E receptor-mediated LDL fractional catabolic rates (FCR) were significantly lower in animals fed the PK diet (P < 0.01) and LDL apoB flux rates were reduced (P < 0.01) in animals fed the BT diet. A correlation was found between plasma LDL levels and receptor-mediated LDL catabolism (r = -0.66, P < 0.01). A higher apoB/E receptor number (Bmax), determined by in vitro LDL binding to guinea pig hepatic membranes, was observed for animals fed BT versus PK or PO diets and Bmax values were significantly correlated with plasma LDL levels (r = -0.776, P < 0.001). These results indicate that saturated fatty acids of varying chain length have differential effects on hepatic apoB/E receptor expression and on LDL apoB flux rates which in part account for differences in plasma LDL cholesterol levels of guinea pigs fed these saturated fats.

Keyword: hyperlipedemia

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as , nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: hyperlipedemia

Palmitate-induced beta-cell dysfunction is associated with excessive NO production and is reversed by thiazolidinedione-mediated inhibition of GPR40 transduction mechanisms.

Type 2 diabetes often displays . We examined palmitate effects on pancreatic islet function in relation to FFA receptor GPR40, NO generation, insulin release, and the PPARgamma agonistic thiazolidinedione, rosiglitazone.Rosiglitazone suppressed acute palmitate-stimulated GPR40-transduced PI hydrolysis in HEK293 cells and insulin release from MIN6c cells and mouse islets. Culturing islets 24 h with palmitate at 5 mmol/l glucose induced beta-cell iNOS expression as revealed by confocal microscopy and increased the activities of ncNOS and iNOS associated with suppression of glucose-stimulated insulin response. Rosiglitazone reversed these effects. The expression of iNOS after high-glucose culturing was unaffected by rosiglitazone. Downregulation of GPR40 by antisense treatment abrogated GPR40 expression and suppressed palmitate-induced iNOS activity and insulin release.We conclude that, in addition to mediating acute FFA-stimulated insulin release, GPR40 is an important regulator of iNOS expression and dysfunctional insulin release during long-term exposure to FFA. The adverse effects of palmitate were counteracted by rosiglitazone at GPR40, suggesting that thiazolidinediones are beneficial for beta-cell function in hyperlipidemic type 2 diabetes.

Keyword: hyperlipedemia

Metabolism of glucose and fatty by leukocytes from patients with endogenous hypertriglyceridemia.

The metabolic abnormalities responsible for endogenous hypertriglyceridemia have not been defined. Some in vivo studies have suggested that excessive triglyceride production is the cause of this defect. In an attempt to obtain direct evidence concerning this mechanism, we have compared in vitro the metabolism of radioactive glucose and palmitate by leukocytes from patients with endogenous hypertriglyceridemia and normal subjects. Leukocytes from the patients incorporated 9.82 plus or minus 1.7 (S.E.M.) nanomoles of glucose into cellular lipid per 10-8 cells per hour. When the cell lipid extract was sugjected to mild alkaline hydrolysis, 92 per cent of the glucose radioactivity was recovered in the glycerol backbone of the lipid esters. Comparison of specific yields of CO2 from glucose labeled in the 1- or 6- position revealed that 0.53 plus or minus 0.02 per cent was metabolized via the pentose cycle. The leukocytes from hypertriglyceridemic persons incorproated 140 plus or minus 6.9 nanomoles of [1-14C]-palmitate per 10-8 cells per hour. Eighty-four per cent of the radioactivity was in triglycerides and 14 per cent in phospholipids. The major phospholipid into which palmitate was incorporated was phosphatidyl choline. The leukocytes oxidized palmitate at a rate of 2.88 plus or minus 0.23 nanomoles per 10-8 cells per hour. There were no differences in any of the above values between leukocytes from hypertriglyceridemic patients and normal subjects. Likewise, there was no correlation between the plasma triglyceride concentration and glucose or palmitate incorporation into triglycerides. To the extent that leukocytes reflect systemic metabolic processes, these data provide no support for the interpretation that the mechanism of the plasma triglyceride elevation is excessive biosynthesis.

Keyword: hyperlipedemia

Palmitate differentially regulates the polarization of differentiating and differentiated macrophages.

The tissue accumulation of M1 macrophages in patients with metabolic diseases such as obesity and type 2 diabetes mellitus has been well-documented. Interestingly, it is an accumulation of M2 macrophages that is observed in the adipose, liver and lung tissues, as well as in the circulation, of patients who have had major traumas such as a burn injury or sepsis; however, the trigger for the M2 polarization observed in these patients has not yet been identified. In the current study, we explored the effects of chronic palmitate and high glucose treatment on macrophage differentiation and function in murine bone-marrow-derived macrophages. We found that chronic treatment with palmitate decreased phagocytosis and HLA-DR expression in addition to inhibiting the production of pro-inflammatory cytokines. Chronic palmitate treatment of bone marrows also led to M2 polarization, which correlated with the activation of the peroxisome proliferator-activated receptor-γ signalling pathway. Furthermore, we found that chronic palmitate treatment increased the expression of multiple endoplasmic reticulum (ER) stress markers, including binding immunoglobulin protein. Preconditioning with the universal ER stress inhibitor 4-phenylbutyrate attenuated ER stress signalling and neutralized the effect of palmitate, inducing a pro-inflammatory phenotype. We confirmed these results in differentiating human macrophages, showing an anti-inflammatory response to chronic palmitate exposure. Though alone it did not promote M2 polarization, hyperglycaemia exacerbated the effects of palmitate. These findings suggest that the dominant accumulation of M2 in adipose tissue and liver in patients with critical illness may be a result of and hyperglycaemia, both components of the hypermetabolism observed in critically ill patients.© 2015 John Wiley & Sons Ltd.

Keyword: hyperlipedemia

[How surplus of fatty in food initiates hypertriglyceridemia, increases cholesterol of low density lipoproteins, triggers atherosclerosis and develops atheromatosis.]

In phylogenesis, the first transfer of all fatty acids to cells is implemented by high density lipoproteins. Later, unsaturated and polyene fatty acids are transferred to cell by low density lipoproteins. The insulin-depended cells absorb saturated fatty , oleic mono-unsaturated fatty and of the same name triglycerides in very low density lipoproteins. The hepatocytes secrete , oleic and linoleic very low density lipoproteins separately. In blood, under hydrolysis of triglycerides, cells absorb ligand and oleic very low density lipoproteins by force of апоЕ/В-100 endocytosis; they are not transformed into low density lipoproteins. The saturated fatty acids in the form of polyether of cholesterol turn into linoleic very low density lipoproteins from high density lipoproteins at impact of protein transferring polyene ethers of cholesterol. They transform very low density lipoproteins into low density lipoproteins of the same name; the cells absorb them by force of апоЕ/В-100 endocytosis. In physiological sense, amount of oleic very low density lipoproteins are always more than of very low density lipoproteins. Under syndrome of insulin-resistance there is no transformation of saturated fatty synthesized from glucose in vivo into oleic mono-saturated fatty . The hepatocytes secrete into blood mainly very low density lipoproteins which amount exceeds oleic very low density lipoproteins. Under slow hydrolysis in blood, main mass of very low density lipoproteins becomes low density lipoproteins. These very lipoproteins initiate , increase content of cholesterol of cholesterol-low density lipoproteins, lower cholesterol-high density lipoproteins, decrease bio-availability of polyene fatty acids for cells, trigger development of atherosclerosis and formation of atheromatosis in intima of arteries. The aphysiologic effect of surplus of saturated fatty in vivo and triglycerides of the same name can\'t be eliminated under increasing of content of ω-3 polyene fatty acids in food and effect of statines. All this is to be rationally applied in prevention of hypertriglyceridemia, atherosclerosis, atheromatosis of coronary arteries, ischemic heart disease and myocardium infarction.

Keyword: hyperlipedemia

Lipid analysis of Coffea arabica Linn. beans and their possible hypercholesterolemic effects.

Lipid composition of green Coffea arabica beans is reported with special emphasis on fatty composition. Triacylglycerols were found to be the major lipid constituents of the coffee oil along with sterol esters, sterols/triterpene alcohol, hydrocarbons and the hydrolyzed products of triacylglycerols as the minor components. Fatty composition of total oil, neutral lipids, polar lipids and pure triacylglycerols showed the presence of fatty acids of C14, C16, C18, and C20 carbon chains. and linoleic acids were the major fatty acids and comprise about 38.7 and 35.9% respectively. Pancreatic lipase hydrolysis revealed that the linoleoyl and palmitoyl moieties are preferentially esterified at the Sn-2 and Sn-1,3 positions of triacylglycerols respectively. The presence of high amounts of at Sn-1,3 position in coffee oil may be partly responsible for its hypercholesterolemic effects.

Keyword: hyperlipedemia

Cholesterolaemic effect of in relation to other dietary fatty acids.

The effect of dietary intake of high levels in combination with other fatty acids in normal subjects was assessed. (10% of energy) was fed in conjunction with decreasing levels of linoleic to determine if a threshold level of linoleic prevented from being hypercholesterolaemic. Healthy subjects received each of the diet treatments for 21 days, followed by washout periods of 7 days. In a second experiment, the effect of exchanging for trans fatty acids on plasma lipoprotein cholesterol levels and on rates for endogenous synthesis of cholesterol in normal subjects was investigated. Diet treatment lasted for 30 days. On day 30 of each diet treatment, a priming dose of deuterium was consumed, followed by a subsequent blood sample at 24 h. Blood cholesterol fractions were isolated and analysed by isotope ratio mass spectrometry to measure cholesterol fractional synthetic rates. In the first experiment, total plasma cholesterol levels increased as the percentage of linoleic decreased. The data indicated that high levels of were not hypercholesterolaemic if intake of linoleic was greater than 4.5% of energy. When the diet contained trans fatty acids plasma total and low-density lipoprotein-cholesterol increased and cholesterol synthesis increased with a decrease in high-density lipoprotein-cholesterol.

Keyword: hyperlipedemia

Plasma metabolic biomarkers for syndrome of phlegm and blood stasis in and atherosclerosis.

To explore the plasma metabolite profiles in patients with the syndrome of phlegm and blood stasis in and atherosclerosis (As), and to search for the metabolic biomarkers of the syndrome.The plasma metabolite profiles of 31 patients with the syndrome of phlegm and blood stasis in and As, 6 patients with syndromes without phlegm and blood stasis, and 10 healthy subjects were analyzed by gas chromatography-mass spectrometry (GC-MS). Partial least squares-discriminant analyses (PLS-DA) were used to carry out the pattern-recognition analyses of the data. The plasma metabolic biomarkers of patients were obtained by variable importance plot value (VIP value) and Student\'s t-test. The structures of biomarkers were defined by the National Institute of Standards and Technology (NIST) database.PLS-DA score plots of plasma metabolomes did not show overlap between the phlegm-blood stasis syndrome group and syndromes without phlegm and blood stasis group, whereas significant differences in the concentrations in the plasma of 5 metabolites were found (P < 0.05). They were identified as urine, isoleucine, glucuronic , and glycerol by searching in NIST database. The concentrations of four metabolites in the plasma of patients with syndrome of phlegm and blood stasis were higher than those with syndromes without phlegm and blood stasis, whereas the glycerol concentration was lower.Compared with patients with syndromes without phlegm and blood stasis, five metabolites showed abnormal levels in patients with the syndrome of phlegm and blood stasis. These metabolites could be diagnostic and prognostic biomarkers.

Keyword: hyperlipedemia

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: hyperlipedemia

[Fatty metabolism in primary hyperlipoproteinemia (HLP) (author\'s transl)].

The fatty spectra of cholesterol ester and triglyceride fractions separated by thin-layer chromatography were analyzed by gas chromatography in the sera of 252 patients with primary HLP (with 53 cases of type IIa, 48 cases of type IIb, and 151 cases of type IV) and 60 healthy persons. Showing significantly increased values in the cholesterol ester fraction for HLP of types IIa and IIb as well as in the triglyceride fraction for HLP of types IIb and IV were , stearic , palmitoleic , oleic , and eicosatrienic . Significantly reduced percentages were obtained for linoleic , linolenic , arachidonic , and eicosapentaenic in the same fractions for these types. Possible causes include disturbances of the fatty metabolism in the liver, increased selective reesterification of fatty acids of the lipolytic process, and partially altered LCAT activity.

Keyword: hyperlipedemia

[Laurine fatty acids, medium fatty acids and triglycerides, , resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: hyperlipedemia

Palm olein and olive oil cause a higher increase in postprandial lipemia compared with lard but had no effect on plasma glucose, insulin and adipocytokines.

Postprandial lipemia impairs insulin sensitivity and triggers the pro-inflammatory state which may lead to the progression of cardiovascular diseases. A randomized, crossover single-blind study (n = 10 healthy men) was designed to compare the effects of a high-fat load (50 g fat), rich in from both plant (palm olein) or animal source (lard) versus an oleic -rich fat (virgin olive oil) on lipemia, plasma glucose, insulin and adipocytokines. Serum triacylglycerol (TAG) concentrations were significantly lower after the lard meal than after the olive oil and palm olein meals (meal effect P = 0.003; time effect P < 0.001). The greater reduction in the plasma non-esterified free fatty acids levels in the lard group compared to the olive oil meal was mirrored by the changes observed for serum TAG levels (P < 0.05). The magnitude of response for plasma glucose, insulin and adipocytokines [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and leptin] were not altered by the type of dietary fats. A significant difference in plasma IL-1β was found over time following the three high fat loads (time effect P = 0.036). The physical characteristics and changes in TAG structure of lard may contribute to the smaller increase in postprandial lipemia compared with palm olein. A high fat load but not the type of fats influences concentrations of plasma IL-1β over time but had no effect on other pro-inflammatory markers tested in the postprandial state.

Keyword: hyperlipedemia

[Dynamic study of the changes in the composition of fatty acids in serum lipid fractions under a diet rich in polyunsaturated fatty acids].

Appropriate adjustment of diet and specially reduction of the saturated fatty acids content is the cornerstone of correcting many . What we have tried in our study was first to show the evolution of the changes in the serum\'s fatty acids composition under a rich polyunsaturated fatty acids diet, second to find an index proving that the diet had correctly been taken. Chromatographic analysis were performed after ultracentrifugal separation of the lipoprotein fractions. Seven volunteers, non fat, and normolipemics were selected. Samples were collected every three or five days before and during the diet. The whole test lasted twenty days. What had been shown is that the linoleic content increased but the one of the oleic decreased in all lipoprotein fractions. Those changes are bestly illustrated by using the (formula: see text) ratio as a kinetic index. The triglycerid fraction modifications are faster than the phospholipid\'s one. Concerning the other fatty acids, the modifications are less spectacular : the decrease in every lipids fractions and the palmitoleic has a significant decrease only in the esterified cholesterol. And the stearic has no variation at least significatively.

Keyword: hyperlipedemia

Lipidomics Biomarkers of Diet-Induced and Its Treatment with Poria cocos.

is a major cause of atherosclerotic cardiovascular disease. Poria cocos (PC) is a medicinal product widely used in Asia. This study was undertaken to define the alterations of lipid metabolites in rats fed a high-fat diet to induce and to explore efficacy and mechanism of action of PC in the treatment of diet-induced . Plasma samples were then analyzed using UPLC-HDMS. The untreated rats fed a high-fat diet exhibited significant elevation of plasma triglyceride and total and low-density lipoprotein (LDL) cholesterol concentrations. This was associated with marked changes in plasma concentrations of seven fatty acids (, hexadecenoic , hexanoylcarnitine, tetracosahexaenoic , cervonoyl ethanolamide, 3-hydroxytetradecanoic , and 5,6-DHET) and five sterols [cholesterol ester (18:2), cholesterol, hydroxytestosterone, 19-hydroxydeoxycorticosterone, and cholic ]. These changes represented disorders of biosynthesis and metabolism of the primary bile acids, steroids, and fatty acids and mitochondrial fatty elongation pathways in diet-induced . Treatment with PC resulted in significant improvements of and the associated abnormalities of the lipid metabolites.

Keyword: hyperlipedemia

Hamburger high in total, saturated and trans-fatty acids decreases HDL cholesterol and LDL particle diameter, and increases TAG, in mildly hypercholesterolaemic men.

The consumption of high-fat hamburger enriched with SFA and trans-fatty acids may increase risk factors for coronary vascular disease, whereas hamburger enriched with MUFA may have the opposite effect. Ten mildly hypercholesterolaemic men consumed five, 114 g hamburger patties per week for two consecutive phases. Participants consumed high-SFA hamburger (MUFA:SFA = 0.95; produced from pasture-fed cattle) for 5 weeks, consumed their habitual diets for 3 weeks and then consumed high-MUFA hamburger (MUFA:SFA = 1.31; produced from grain-fed cattle) for 5 weeks. These MUFA:SFA ratios were typical of ranges observed for retail ground beef. Relative to habitual levels and levels during the high-MUFA phase, the high-SFA hamburger: increased plasma , palmitoleic and TAG (P < 0.01); decreased HDL cholesterol (HDL-C) and LDL particle diameter percentile distributions (P < 0.05); and had no effect on LDL cholesterol or plasma glucose (P>0.10). Plasma palmitoleic was positively correlated with TAG (r 0.90), VLDL cholesterol (r 0.73) and the LDL:HDL ratio (r 0.45), and was negatively correlated with plasma HDL-C (r - 0.58), whereas plasma , stearic and oleic acids were negatively correlated with LDL particle diameter (all P

Keyword: hyperlipedemia

Elevated circulating stearic leads to a major lipotoxic effect on mouse pancreatic beta cells in via a miR-34a-5p-mediated PERK/p53-dependent pathway.

Serum stearic (C18:0) is elevated in individuals with and type 2 diabetes. However, the lipotoxicity induced by increased stearic in beta cells has not been well described. This study aimed to examine the adverse effects of stearic on beta cells and the potential mechanisms through which these are mediated.Three groups of C57BL/6 mice were fed a normal diet or a high-stearic-/high- diet for 24\xa0weeks, respectively. The microRNA (miR) profiles of islets were determined by microarray screening. Islet injury was detected with co-staining using the TUNEL assay and insulin labelling. A lentiviral vector expressing anti-miRNA-34a-5p oligonucleotide (AMO-34a-5p) was injected into mice via an intraductal pancreatic route.In both mouse islets and cultured rat insulinoma INS-1 cells, stearic exhibited a stronger lipotoxic role than other fatty acids, owing to repression of B cell CLL/lymphoma 2 (BCL-2) and BCL-2-like 2 (BCL-W) by stearic stimulation of miR-34a-5p. The stearic--induced lipotoxicity and reduction in insulin secretion were alleviated by AMO-34a-5p. Further investigations in INS-1 cells revealed that p53 was involved in stearic--induced elevation of miR-34a-5p, owing in part to activation of protein kinase-like endoplasmic reticulum kinase (PERK). Conversely, silencing PERK alleviated stearic--induced p53, miR-34a-5p and lipotoxicity.These findings provide new insight for understanding the molecular mechanisms underlying not only the deleterious impact of stearic--induced lipotoxicity but also apoptosis in beta cells and progression to type 2 diabetes.

Keyword: hyperlipedemia

Serum lipoprotein fatty patterns in various types of familiar combined .

The relative content of various fatty acids in serum lipoproteins was determined in patients with type IIa (38), IIb (49) and IV (77) of hyperlipoproteinemia and compared with 52 controls. Significant changes were found in hyperlipoproteinemia associated with hypertriglyceridemia (type IV) but not in "pure" hypercholesterolemia (type IIa). In all lipoprotein fractions (VLDL, LDL, HDL) in type IV of hyperlipoproteinemia the increased oleic and linolenic proportions were found, while proportions of linoleic, arachidonic and docosahexaenoic acids were decreased. The saturated fatty acids (myristic, and stearic) were found increased in LDL. Linear regression analysis has shown positive correlation between the content of arachidonic and docosahexaenoic acids in HDL and LDL and the serum levels of total HDL-cholesterol, HDL2-cholesterol, HDL3-cholesterol and ApoA1, while a negative correlation between these fatty acids and serum triglycerides level appeared. These findings can be explained partly by increased content of triglycerides and free fatty acids in lipoproteins. Possible differences concerning mechanisms of accelaration of atherogenesis in various types of are discussed.

Keyword: hyperlipedemia

Effects of Plant Oil Interesterified Triacylglycerols on Lipemia and Human Health.

The position of the fatty acids (-1, -2 and -3) (stereospecific numbering ()) in triacylglycerol (TAG) molecules produces a characteristic stereospecificity that defines the physical properties of the fats and influences their absorption, metabolism and uptake into tissues. Fat interesterification is a process that implies a positional distribution of fatty acids (FAs) within the TAG molecules, generating new TAG species, without affecting the FA - natural balance. The interesterified (IE) fats, frequently used in the food industry comprise fats that are rich in long-chain saturated FAs, such as (16:0) and stearic (18:0). Within the interesterified fats, a critical role is played by FA occupying the -2 position; in fact, the presence of an unsaturated FA in this specific position influences early metabolic processing and postprandial clearance that in turn could induce atherogenesis and thrombogenesis events. Here, we provide an overview on the role of TAG structures and interesterified and stearic -rich fats on fasting and postprandial lipemia, focusing our attention on their physical properties and their effects on human health.

Keyword: hyperlipedemia

Platelet deposition at high shear rates is enhanced by high plasma cholesterol levels. In vivo study in the rabbit model.

We have studied the effects of high plasma cholesterol levels on platelet-vessel wall interactions under high shear rate conditions typical of the apex of stenotic arteries (2,600 sec-1). Hypercholesterolemia was induced by feeding rabbits a 0.5% cholesterol-rich diet for 60 days. Platelet deposition was studied by use of an annular perfusion chamber and de-endothelialized abdominal rabbit aortas as substrates. After ingestion of the atherogenic diet, the experimental group of animals developed severe hypercholesterolemia, platelets became more fluid as determined by steady-state fluorescence anisotropy (p less than 0.05), and red blood cell deformability was decreased (p less than 0.001) when compared with normal controls. The fatty composition of platelet membranes showed an increase in the percentage of the long-chain saturated fatty acids (, C16:0, and stearic, C18:0) that may account for the lower polyunsaturated/saturated fatty ratio observed in the hyperlipemic animals. Total platelet deposition was significantly increased (p less than 0.05) in the hyperlipemic group as compared with the control group at 5 minutes\' perfusion time, becoming less evident at 20 minutes\' perfusion time. Our results suggest that the presence of may contribute to acute thrombosis by enhancing platelet-vessel wall interaction.

Keyword: hyperlipedemia

METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, insulin resistance, and diabetes. Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: hyperlipedemia

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the liver of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in liver cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the liver steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and liver lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: hyperlipedemia

is associated with increased insulin-mediated glucose metabolism, reduced fatty metabolism and normal blood pressure in transgenic mice overexpressing human apolipoprotein C1.

Insulin resistance for glucose metabolism is associated with and high blood pressure. In this study we investigated the effect of primary on basal and insulin-mediated glucose and on non-esterified fatty (NEFA) metabolism and mean arterial pressure in hyperlipidaemic transgenic mice overexpressing apolipoprotein C1 (APOC1). Previous studies have shown that APOC1 transgenic mice develop primarily because of an impaired hepatic uptake of very low density lipoprotein (VLDL).Basal and hyperinsulinaemic (6 mU.kg-1.min-1), euglycaemic (7 mmol/l) clamps with 3(-)3H-glucose or 9,10(-)3H- infusions and in situ freeze clamped tissue collection were carried out.The APOC1 mice showed increased basal plasma cholesterol, triglyceride, NEFA and decreased glucose concentrations compared with wild-type mice (7.0 +/- 1.2 vs 1.6 +/- 0.1, 9.1 +/- 2.3 vs 0.6 +/- 0.1, 1.9 +/- 0.2 vs 0.9 +/- 0.1 and 7.0 +/- 1.0 vs 10.0 +/- 1.1 mmol/l, respectively, p < 0.05). Basal whole body glucose clearance was increased twofold in APOC1 mice compared with wild-type mice (18 +/- 2 vs 10 +/- 1 ml.kg-1.min-1, p < 0.05). Insulin-mediated whole body glucose uptake, glycolysis (generation of 3H2O) and glucose storage increased in APOC1 mice compared with wild-type mice (339 +/- 28 vs 200 +/- 11; 183 +/- 39 vs 128 +/- 17 and 156 +/- 44 vs 72 +/- 17 mumol.kg-1.min-1, p < 0.05, respectively), corresponding with a twofold to threefold increase in skeletal muscle glycogenesis and de novo lipogenesis from 3-(3)H-glucose in skeletal muscle and adipose tissue (p < 0.05). Basal whole body NEFA clearance was decreased threefold in APOC1 mice compared with wild-type mice (98 +/- 21 vs 314 +/- 88 ml.kg-1.min-1, p < 0.05). Insulin-mediated whole body NEFA uptake, NEFA oxidation (generation of 3H2O) and NEFA storage were lower in APOC1 mice than in wild-type mice (15 +/- 3 vs 33 +/- 6; 3 +/- 2 vs 11 +/- 4 and 12 +/- 2 vs 22 +/- 4 mumol.kg-1.min-1, p < 0.05) in the face of higher plasma NEFA concentrations (1.3 +/- 0.3 vs 0.5 +/- 0.1 mmol/l, p < 0.05), respectively. Mean arterial pressure and heart rate were similar in APOC1 vs wild-type mice (82 +/- 4 vs 85 +/- 3 mm Hg and 459 +/- 14 vs 484 +/- 11 beats.min-1).1) Hyperlipidaemic APOC1 mice show reduced NEFA and increased glucose metabolism under both basal and insulin-mediated conditions, suggesting an intrinsic defect in NEFA metabolism. Primary alone in APOC1 mice does not lead to insulin resistance for glucose metabolism and high blood pressure.

Keyword: hyperlipedemia

A meal rich in oleic beneficially modulates postprandial sICAM-1 and sVCAM-1 in normotensive and hypertensive hypertriglyceridemic subjects.

This study investigated whether subjects with permanent activated endothelium have altered soluble forms of intercellular adhesion molecule 1 (sICAM-1) and vascular cell adhesion molecule 1 (sVCAM-1) postprandial response to a high-fat meal and whether this phenomenon is modulated by the nature of dietary fats. Twenty-eight hypertriglyceridemic (14 normotensives and 14 hypertensives) and 14 healthy male subjects were placed in a randomized and crossover design on diets enriched in refined olive oil (ROO) or high- sunflower oil (HPSO) for a 1-week lead-in period. Thereafter, subjects ate the corresponding fat-rich meal as a breakfast and underwent sampling hourly for 8 h. Plasma triglycerides (TG), sICAM-1 and sVCAM-1 were assayed. sICAM-1 and sVCAM-1 postprandial peak levels were significantly higher and occurred later in hypertriglyceridemic subjects (all P<.001) compared with healthy subjects. ROO meal resulted in smaller areas under the curve for sICAM-1 and sVCAM-1 in hypertriglyceridemic (normotensive and hypertensive) and healthy subjects compared to HPSO meal. Hypertension did not aggravate the postprandial response of TG, sICAM-1 and sVCAM-1. We conclude that the challenge of a meal with ROO appears to have a significant postprandial benefit on sICAM-1 and sVCAM-1 as surrogate markers of endothelial activation and vascular inflammation in healthy and more importantly in hypertriglyceridemic (normotensive and hypertensive) subjects.

Keyword: hyperlipedemia

Cyclosporine A and treatment synergistically induce cytotoxicity in HepG2 cells.

Immunosuppressant cyclosporine A (CsA) treatment can cause severe side effects. Patients taking immunosuppressant after organ transplantation often display and obesity. Elevated levels of free fatty acids have been linked to the etiology of metabolic syndromes, nonalcoholic fatty liver and steatohepatitis. The contribution of free fatty acids to CsA-induced toxicity is not known. In this study we explored the effect of on CsA-induced toxicity in HepG2 cells. CsA by itself at therapeutic exposure levels did not induce detectible cytotoxicity in HepG2 cells. Co-treatment of and CsA resulted in a dose dependent increase in cytotoxicity, suggesting that fatty could sensitize cells to CsA-induced cytotoxicity at the therapeutic doses of CsA. A synergized induction of caspase-3/7 activity was also observed, indicating that apoptosis may contribute to the cytotoxicity. We demonstrated that CsA reduced cellular oxygen consumption which was further exacerbated by , implicating that impaired mitochondrial respiration might be an underlying mechanism for the enhanced toxicity. Inhibition of c-Jun N-terminal kinase (JNK) attenuated and CsA induced toxicity, suggesting that JNK activation plays an important role in mediating the enhanced /CsA-induced toxicity. Our data suggest that elevated FFA levels, especially saturated FFA such as , may be predisposing factors for CsA toxicity, and patients with underlying diseases that would elevate free fatty acids may be susceptible to CsA-induced toxicity. Furthermore, /obesity resulting from immunosuppressive therapy may aggravate CsA-induced toxicity and worsen the outcome in transplant patients.Copyright © 2012 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Variations in serum cholesteryl ester transfer and phospholipid transfer activities in healthy women and men consuming diets enriched in lauric, or oleic acids.

Cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP) activities were measured in sera from 32 normolipidemic women and men consuming diets enriched in lauric, , or oleic acids. Serum CETP activity, measured as the rate of radiolabeled cholesteryl esters transferred from HDL toward serum apo B-containing lipoproteins, was higher with the diet (25.1+/-2.5%) than with the lauric (23.7+/-2.4%) and the oleic (24.0+/-2.7%) diets (P = 0.0028 and 0.0283, respectively). CETP mass concentrations, as measured with an enzyme-linked immunosorbent assay were increased after the lauric diet (2.57+/-0.63 mg/l) and the diet (2.49+/-0.64 mg/l) as compared with the oleic diet (2.34+/-0.45 mg/l) (P = 0.0035 and 0.0249, respectively). In contrast with CETP, serum PLTP activity, as measured as the rate of radiolabeled phosphatidylcholine transferred from liposomes toward serum HDL, was significantly higher with the lauric diet (23.5+/2.6%) than with the diet (22.5+/-2.5%) (P = 0.0013), while no significant differences were noted when comparing the saturated diets versus the oleic diet (23.0+/-2.3%). No significant alterations in the mean apparent diameter of LDL, and in the relative proportions of individual HDL subpopulations were observed from one dietary period to another. Nevertheless, lipid transfer activities correlated significantly with the relative abundance of HDL2b, HDL2a, HDL3b, and HDL3c, with opposite tendencies being observed for cholesteryl ester transfer and phospholipid transfer activities. In general, serum CETP activity correlated negatively with HDL cholesterol, but positively with triglyceride concentrations after the dietary interventions, and the relations with serum lipids were just the opposite for PLTP activity. In addition, CETP and PLTP activities correlated negatively when subjects consumed the standardized diets (P < 0.05 in all cases), but not when subjects consumed their habitual diet. It is concluded that serum lipid transfer activities in normolipidemic subjects can be significantly affected by the fatty content of the diet, with differential effects on CETP and PLTP activities.

Keyword: hyperlipedemia

interferes with energy metabolism balance by adversely switching the SIRT1-CD36-fatty pathway to the PKC zeta-GLUT4-glucose pathway in cardiomyoblasts.

Metabolic regulation is inextricably linked with cardiac function. Fatty metabolism is a significant mechanism for creating energy for the heart. However, cardiomyocytes are able to switch the fatty acids or glucose, depending on different situations, such as ischemia or anoxia. Lipotoxicity in obesity causes impairments in energy metabolism and apoptosis in cardiomyocytes. We utilized the treatment of H9c2 cardiomyoblast cells (PA) as a model for to investigate the signaling mechanisms involved in these processes. Our results show PA induces time- and dose-dependent lipotoxicity in H9c2 cells. Moreover, PA enhances cluster of differentiation 36 (CD36) and reduces glucose transporter type 4 (GLUT4) pathway protein levels following a short period of treatment, but cells switch from CD36 back to the GLUT4 pathway after during long-term exposure to PA. As sirtuin 1 (SIRT1) and protein kinase Cζ (PKCζ) play important roles in CD36 and GLUT4 translocation, we used the SIRT1 activator resveratrol and si-PKCζ to identify the switches in metabolism. Although PA reduced CD36 and increased GLUT4 metabolic pathway proteins, when we pretreated cells with resveratrol to activate SIRT1 or transfected si-PKCζ, both were able to significantly increase CD36 metabolic pathway proteins and reduce GLUT4 pathway proteins. High-fat diets affect energy metabolism pathways in both normal and aging rats and involve switching the energy source from the CD36 pathway to GLUT4. In conclusion, PA and high-fat diets cause lipotoxicity in vivo and in vitro and adversely switch the energy source from the CD36 pathway to the GLUT4 pathway.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Activation of peroxisome proliferator-activated receptor-α (PPARα) in proximal intestine improves postprandial lipidemia in obese diabetic KK-Ay mice.

Postprandial lipidemia is a risk factor for cardiovascular diseases. Thus, the suppression of postprandial lipidemia is valuable for disease management. Peroxisome proliferator-activated receptor- (PPAR ) is a key regulator in the lipid metabolism of peripheral tissues such as the liver and skeletal muscle, whose activation enhances fatty oxidation and decreases circulating lipid level. Recently, we have shown that bezafibrate, an agonistic compound for PPAR , suppresses post-prandial lipidemia by enhancing fatty oxidation in intestinal epithelial cells under physiological conditions. However, it was not elucidated whether the effect of PPAR on postprandial lipidemia is also observed under obese conditions, which change lipid metabolisms in various tissues and cells. Here, we observed that bezafibrate enhanced fatty oxidation in intestinal epithelial cells of obese diabetic KK-Ay mice. Bezafibrate treatment increased the mRNA expression levels of fatty oxidation-related genes, which are targets of PPAR , and enhanced CO2 production from [14C]-. The bezafibrate-treated mice showed the suppression of increasing serum triacylglyceride level after the oral administration of olive oil. Moreover, the effects of bezafibrate on mRNA expression and fatty oxidation were shown in only the proximal intestinal epithelial cells. These findings indicate that PPAR activation suppresses postprandial lipidemia under obese conditions through the enhancement of fatty oxidation, and that only the proximal intestine con-tributes to the effects in mice, suggesting that intestinal PPAR can be a target for prevention of obese-induced postprandial lipidemia.© 2013 Asian Oceanian Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.

Keyword: hyperlipedemia

MEDICA 16 inhibits hepatic acetyl-CoA carboxylase and reduces plasma triacylglycerol levels in insulin-resistant JCR: LA-cp rats.

Intracellular triacylglycerol (TG) content of liver and skeletal muscle contributes to insulin resistance, and a significant correlation exists between TG content and the development of insulin resistance. Because acetyl-CoA carboxylase (ACC) is the rate-limiting enzyme for liver fatty biosynthesis and a key regulator of muscle fatty oxidation, we examined whether ACC plays a role in the accumulation of intracellular TG. We also determined the potential role of 5\'-AMP-activated protein kinase (AMPK) in this process, since it can phosphorylate and inhibit ACC activity in both liver and muscle. TG content, ACC, and AMPK were examined in the liver and skeletal muscle of insulin-resistant JCR:LA-cp rats during the time frame when insulin resistance develops. At 12 weeks of age, there was a threefold elevation in liver TG content and a sevenfold elevation in skeletal muscle TG content. Hepatic ACC activity was significantly elevated in 12-week-old JCR:LA-cp rats compared with lean age-matched controls (8.75 +/- 0.53 vs. 3.30 +/- 0.18 nmol. min(-1). mg(-1), respectively), even though AMPK activity was also increased. The observed increase in hepatic ACC activity was accompanied by a 300% increase in ACC protein expression. There were no significant differences in ACC activity, ACC protein expression, or AMPK activity in the skeletal muscle of the 12-week JCR:LA-cp rats. Treatment of 12-week JCR:LA-cp rats with MEDICA 16 (an ATP-citrate lyase inhibitor) resulted in a decrease in hepatic ACC and AMPK activities, but had no effect on skeletal muscle ACC and AMPK. Our data suggest that alterations in ACC or AMPK activity in muscle do not contribute to the development of insulin resistance. However, increased liver ACC activity in the JCR:LA-cp rat appears to contribute to the development of lipid abnormalities, although this increase does not appear to occur secondary to a decrease in AMPK activity.

Keyword: hyperlipedemia

[Fatty and lipid peroxidation in human atherosclerosis].

Plasma fatty acids and lipid peroxidation were studied in human atherosclerosis. Analysis of fatty acids in 16 controls and 32 hyperlipidemic patients showed, in the latter, a decrease in saturated fatty acids, especially and stearic acids, and an increase in unsaturated fatty acids, especially arachidonic . Compared to hyperlipidemic patients without arterial injury, patients with arterial injury exhibit a significant increase in malonaldehyde (MDA). In the former, MDA concentrations are significantly increased compared to controls. Therefore, peroxidation of unsaturated fatty acids may have a deleterious effect on arteries in atheroma, through the release of toxic endoperoxydes and the metabolization of arachidonic into thromboxane, which is a platelet aggregator. Lipid peroxidation can also be demonstrated in other diseases: we found very high MDA concentration in 11 alcoholic patients (alcoholic hepatitis, cirrhosis) and 6 patients with inflammatory conditions such as Crohn disease.

Keyword: hyperlipedemia

Individual responsiveness to a cholesterol-lowering diet in postmenopausal women with moderate hypercholesterolemia.

The efficacy of the step 1 diet in outpatient women with hypercholesterolemia has been debated.Forty-one normotriglyceridemic women whose low-density lipoprotein (LDL) cholesterol levels were 3.62 to 5.17 mmol/L (140 to 200 mg/dL) participated in a two-period outpatient diet counseling study that used a 1-month high-fat, high-saturated fatty period (Hi-Sat diet) and a 4-month low-fat, low-saturated fatty period (step 1 diet). All women were postmenopausal and were not taking hormone replacement therapy. Levels of lipids, lipoproteins, and plasma triglyceride fatty acids were measured five times during the last 2 weeks of each dietary period. Dietary intake was assessed by 7-day food records.The mean reduction in total cholesterol level achieved by the step 1 diet was 0.36 mmol/L (14 mg/dL). The reduction in total cholesterol level was seen in both LDL cholesterol levels (0.28 mmol/L [11 mg/dL]; P < .005) and high-density lipoprotein cholesterol levels (0.08 mmol/L [3 mg/dL]; P = .08). Although individual LDL responsiveness to diet was normally distributed, there was marked variation in response, which could be explained only partially by compliance (change in saturated fat intake, 10%), change in body weight (3%), and an interaction between the content of the plasma and body weight (3%).A step 1 diet lowers total and LDL cholesterol levels in postmenopausal women. A nonsignificant reduction in high-density lipoprotein cholesterol levels was also observed. Since some women achieved LDL cholesterol levels low enough to obviate the need for drug therapy as primary prevention of coronary heart disease, dietary therapy should remain the first step in the management of hypercholesterolemia in postmenopausal women.

Keyword: hyperlipedemia

[Preliminary evaluation of an association of polyunsaturated acids in primary hypertriglyceridemia].

The above study was intended to evaluate changes in triglyceride blood level in subjects treated with a polyunsaturated fatty combination: docosahexanoic (DHA) and eicosapentaenoic (EPA) , compared to changes observed in patients treated with arachidonic . The results obtained show that a significant reduction of triglyceride blood level was achieved and was more marked in patients treated with the DHA-EPA combination compared to those treated with arachidonic only. An increase in the index of fatty unsaturation was observed in all patients without significant differences between treatments.

Keyword: hyperlipedemia

Cholesterol efflux from cells enriched with cholesteryl esters by incubation with hypercholesterolemic monkey low density lipoprotein.

The mass efflux of free and esterified cholesterol was studied in skin fibroblasts loaded with cholesterol by incubation with low density lipoproteins (LDL) isolated from normal or hypercholesterolemic cynomolgus monkeys. Cells incubated with hypercholesterolemic LDL accumulated 2-3 times more cholesteryl ester than did cells incubated with the same amount of normal LDL. Cholesteryl oleate was the principal cholesteryl ester species to accumulate in cells incubated with both normal and hypercholesterolemic LDL. Efflux of this accumulated cholesterol was absolutely dependent on the presence of a cholesterol acceptor in the culture medium. Lipoprotein-deficient serum (LPDS) was the most potent promoter of cholesterol efflux tested, with maximum efflux occurring at LPDS concentrations greater than 1.5 mg protein/ml. Upon addition of efflux medium containing LPDS, there was a reduction in both the free and esterified cholesterol concentration of the cells. Greater than 90% of the cholesteryl esters that were lost from the cells appeared in the culture medium as free cholesterol, indicating that hydrolysis of cholesteryl esters preceded efflux. Efflux was not inhibited by chloroquine, however, suggesting a mechanism independent of lysosomes. Loss of cellular free cholesterol was maximum by 6 hr and changed very little thereafter up to 72 hr. Cholesteryl ester loss from cells decreased in a log linear fashion for efflux periods of 6-72 hr, with an average half-life for cholesteryl ester efflux of 30 hr, but with a range of 20-50 hr, depending upon the specific cell line. The rate of efflux of cellular cholesteryl esters was similar for cells loaded with normal or hypercholesterolemic LDL. In cells loaded with cholesteryl esters, cholesterol synthesis was suppressed and cholesterol esterification and fatty synthesis were enhanced. During efflux, cholesterol synthesis remained maximally suppressed while cholesterol esterification decreased for the first 24 hr of efflux, then plateaued at a level approximately 5-fold higher than control levels, while fatty synthesis was slightly stimulated. There was little difference in the rate of efflux of individual cholesteryl ester species. There was, however, the suggestion that reesterification of cholesterol principally to occurred during efflux. Since the rate of cellular cholesteryl ester efflux was similar regardless of whether the cells had been loaded with cholesterol by incubation with normal LDL or hypercholesterolemic LDL, the greater accumulation of cholesterol in cells incubated with hypercholesterolemic LDL cannot be explained by differences in rates of efflux.-St. Clair, R. W., and M. A. Leight. Cholesterol efflux from cells enriched with cholesteryl esters by incubation with hypercholesterolemic monkey low density lipoprotein.

Keyword: hyperlipedemia

Activation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase during high fat diet feeding.

The liver plays a central role in regulating cholesterol homeostasis. High fat diets have been shown to induce obesity and . Despite considerable advances in our understanding of cholesterol metabolism, the regulation of liver cholesterol biosynthesis in response to high fat diet feeding has not been fully addressed. The aim of the present study was to investigate mechanisms by which a high fat diet caused activation of liver 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) leading to increased cholesterol biosynthesis. Mice were fed a high fat diet (60% kcal fat) for 5weeks. High fat diet feeding induced weight gain and elevated lipid levels (total cholesterol and triglyceride) in both the liver and serum. Despite cholesterol accumulation in the liver, there was a significant increase in hepatic HMG-CoA reductase mRNA and protein expression as well as enzyme activity. The DNA binding activity of sterol regulatory element binding protein (SREBP)-2 and specific protein 1 (Sp1) were also increased in the liver of mice fed a high fat diet. To validate the in vivo findings, HepG2 cells were treated with . Such a treatment activated SREBP-2 as well as increased the mRNA and enzyme activity of HMG-CoA reductase leading to intracellular cholesterol accumulation. Inhibition of Sp1 by siRNA transfection abolished -induced SREBP-2 and HMG-CoA reductase mRNA expression. These results suggest that Sp1-mediated SREBP-2 activation contributes to high fat diet induced HMG-CoA reductase activation and increased cholesterol biosynthesis. This may play a role in liver cholesterol accumulation and hypercholesterolemia.Copyright © 2013 Elsevier B.V. All rights reserved.

Keyword: hyperlipedemia

Inhibitory effects of chondroitin sulfate prepared from salmon nasal cartilage on fat storage in mice fed a high-fat diet.

Chondroitin sulfate is an acidic polymer consisting of repeating D-glucuronic and D-N-acetylgalactosamine units, and the N-acetylgalactosamine is substituted with the sulfate at either the 4\' or 6\' position, with approximately one sulfate being present per disaccharide unit. The present study assessed the effects of chondroitin sulfate on the activity of pancreatic lipase and lipid uptake into brush border membrane vesicles of the rat small intestine in vitro, and on the degree of fat storage induced in mice by the oral administration of a high-fat diet for 8 weeks.Experiments were carried out to clarify whether or not chondroitin sulfate inhibited pancreatic lipase activity in assay systems using triolein emulsified with phosphatidylcholine or gum arabic. In addition, the effects of chondroitin sulfate on lipid absorption by brush border membrane vesicles were examined. Moreover, mice were fed a high-fat diet and treated with chondroitin sulfate for 8 weeks.Chondroitin sulfate dose-dependently inhibited the pancreatic lipase activity in an assay system using triolein emulsified with phosphatidylcholine. In addition, chondroitin sulfate inhibited the uptake into the brush border membrane vesicles of the rat jejunum. Chondroitin sulfate caused the reduction of body weight and parametrial adipose tissue weight, and prevention of fatty liver and in mice fed a high-fat diet.The reduction of fat storage and the antihyperlipidemic action of chondroitin sulfate might be due to the inhibition of small intestinal absorption of dietary fat through the inhibition of pancreatic lipase activity and fatty uptake through brush border membrane.

Keyword: hyperlipedemia

Plasma n-3 fatty acids and the risk of cognitive decline in older adults: the Atherosclerosis Risk in Communities Study.

Plasma fatty acids may affect the risk of cognitive decline in older adults.We prospectively studied the association between plasma fatty acids and cognitive decline in adults aged 50-65 y at baseline and conducted a subgroup analysis.From 1987 through 1989, the Atherosclerosis Risk in Communities (ARIC) Study analyzed plasma fatty acids in cholesteryl esters and phospholipids in whites residing in Minneapolis, MN. From 1990 through 1992 and from 1996 through 1998, 3 neuropsychological tests in the domains of delayed word recall, psychomotor speed, and verbal fluency were administered. We selected cutoffs for statistically reliable cognitive decline in each of these domains and a measure of global cognitive change computed by principal-components analysis. Multivariate logistic regression was conducted. Focusing on n-3 highly unsaturated fatty acids (HUFAs), a subgroup analysis assessed differential association across potential effect modifiers implicated in oxidative stress and increased risk of neurodegenerative disease.In the 2251 study subjects, the risk of global cognitive decline increased with elevated in both fractions and with high arachidonic and low linoleic in cholesteryl esters. Higher n-3 HUFAs reduced the risk of decline in verbal fluency, particularly in hypertensive and dyslipidemic subjects. No significant findings were shown for psychomotor speed or delayed word recall.Promoting higher intakes of n-3 HUFAs in the diet of hypertensive and dyslipidemic persons may have substantial benefits in reducing their risk of cognitive decline in the area of verbal fluency. However, clinical trials are needed to confirm this finding.

Keyword: hyperlipedemia

Antihyperlipidemic Effect, Identification and Isolation of the Lipophilic Components from Artemisia integrifolia.

L (Compositae) is a medicinal and edible plant. To investigate its antihyperlipidemic effect, a crude lipophilic extract and the composing compounds were isolated and fractioned from the petroleum ether extract of aerial parts of using column chromatography on silica gel. The anti- effect was studied in a rat model of acute , which was induced by triton WR-1339. A new compound, integrinol (), together with nine known compounds, namely chamazulene (), acetylenes (E)-2 (), acetylenes (E)-3 (), eugenol (), (), oleic (), linoleic (), linolenic () and 12,13-epoxylinolenic were isolated from the crude lipophilic extract of A. integrifolia. The LD50 value of the crude extract was more than 4g/kg. In Triton WR-1339-induced acute model, the crude lipophilic extract (200 mg/kg) significantly reduced total cholesterol (TC) by 70% ( ≤ 0.01) and triglycerides (TGs) by 94% ( ≤ 0.001). The fractioned compounds, such as chamazulene (1), acetylene-2 (2), and linolenic (9), used at 4 mg/kg dose, also significantly decreased the concentrations of TC (32%, 33% and 64%, respectively) and TGs (48%, 33% and 93%, respectively). These compounds (i.e., chamazulene, acetylenes (E)-2, and linolenic ) were considered to be responsible for the bioactive antihyperlipidemic effect. In conclusion, the crude lipid extract of Artemisia integrifolia L could be used as a potential treatment to avert . Further studies to confirm these results in other models of (e.g., diet-induced obesity) are warranted.

Keyword: hyperlipedemia

Sterol regulatory element-binding protein-1c mediates increase of postprandial stearic , a potential target for improving insulin resistance, in .

Elevated serum free fatty acids (FFAs) levels play an important role in the development of insulin resistance (IR) and diabetes. We investigated the dynamic changes and the underlying regulatory mechanism of postprandial FFA profile in (HLP) and their relation with insulin sensitivity in both humans and mice. We found that serum stearic (SA) is the only fatty that is increased dramatically in the postprandial state. The elevation of SA is due to increased insulin-stimulated de novo synthesis mediated by sterol regulatory element-binding protein-1c (SREBP-1c)/acetyl-CoA carboxylase/fatty synthase/elongation of long-chain fatty family member 6 (ELOVL6) and the elongation of (PA) catalyzed by ELOVL6. Downregulation of SREBP-1c or ELOVL6 by small interfering RNA can reduce SA synthesis in liver and serum SA level, followed by amelioration of IR in HLP mice. However, inhibition of SREBP-1c is more effective in improving IR than suppression of ELOVL6, which resulted in accumulation of PA. In summary, increased postprandial SA is caused by the insulin-stimulated SREBP-1c pathway and elongation of PA in HLP. Reduction of postprandial SA is a good candidate for improving IR, and SREBP-1c is potentially a better target to prevent IR and diabetes by decreasing SA.

Keyword: hyperlipedemia

Elucidation of mechanisms of actions of thymoquinone-enriched methanolic and volatile oil extracts from Nigella sativa against cardiovascular risk parameters in experimental .

Nigella sativa belonging to the Ranunculaceae family has been reported to use for thousands of years as protective and curative traditional medicine against a number of diseases. GC-MS analysis of methanolic extract (ME) and volatile oil (VO) extracted from Nigella sativa seed oil was performed by two different mass spectrometry libraries, WIlEY8 and NIST05s. The cholesterol lowering and antioxidant actions of VO and ME fractions were investigated in atherogenic suspension fed rats.In this study, four groups of male Wistar rats were used: normolipidemic control (NLP-C), hyperlipidemic control (HLP-C), methanolic extract (HLP-ME) and volatile oil treated (HLP-VO) groups for 30 days of duration. P value < 0.05 was assumed as significant data in groups.Administration of atherogenic suspension to male Wistar rats for 30 days resulted in a marked increase of plasma triglycerides and total cholesterol, and significant change in plasma lipoprotein levels along with a decrease in antioxidant arylesterase activity in hyperlipidemic control (HLP-C) group. The oral feeding of 100 mg ME or 20 mg VO per rat/day effectively reduced the plasma triglycerides to near normal level, while high density lipoprotein cholesterol and its subfraction along with arylesterase activity levels were significantly increased. The test fractions elicited a significant decrease in hepatic HMG-CoA reductase activity. The fractions significantly blocked the ex vivo basal and in vitro maximal formation of conjugated diene and malondialdehyde, and lengthened the lag times of low density lipoprotein, small dense low density lipoprotein and large buoyant low density lipoprotein. ME possessing ω-6 linoleic along with active compounds was more effective than VO extract containing thymol and isothymol phenolic antioxidant compounds, thymoquinone phenolic compound common to the both extracts, via reduction in hepatic HMG-CoA reductase activity as well as antioxidant mechanisms.The both extracts especially, ME significantly improve cardiovascular risk parameters in treated rats, and can be used in reactive oxygen species disorders such as cardiovascular diseases.

Keyword: hyperlipedemia

The effect of on lipoprotein cholesterol levels and endogenous cholesterol synthesis in hyperlipidemic subjects.

The present study assesses the effect of high vs. low intakes on plasma lipoprotein cholesterol levels and on rates for endogenous synthesis of cholesterol in healthy and hyperlipidemic subjects. Four diets were formulated to provide combinations of 16:0 at two levels of 18:2n-6. Subjects received each diet treatment for 21 d, followed by washout periods of 21 d. On day 21 of each diet treatment, a fasting blood sample was drawn for lipoprotein determination and to provide a measure of the background level of deuterium. A priming dose of deuterium was consumed and a second blood sample obtained 24 h after the first sample. Isotope ratio mass spectrometry was used to determine the incorporation of deuterium into the newly synthesized cholesterol molecule, and fractional synthetic rates were calculated. Serum total cholesterol and low density lipoprotein-cholesterol was not significantly affected by the high level of 16:0 when diets also contained a high level of 18:2n-6. There was no effect of dietary 16:0 on high density liproprotein-cholesterol at either the high or low levels of intake. The results indicate that 16:0 has no effect on serum lipoprotein profiles in the presence of recommended intakes for 18:2n-6.

Keyword: hyperlipedemia

Biochemical and functional alterations associated with hypercholesterolemia in platelets from hypertensive patients.

Hypercholesterolemia and hypertension are two of the major risk factors associated with increased atherosclerotic vascular disease. An abnormal platelet function is one of the mechanisms proposed to participate in atherogenesis. This study was undertaken to find out whether hypercholesterolemia in hypertensive patients can change platelet lipid composition and reactivity. Twenty-nine untreated hypertensive patients were distributed into 3 age, body mass index and blood pressure-matched groups according to their plasma cholesterol levels (normal, borderline or elevated, group NC, BC and HC respectively). Their platelet lipid composition, cytosolic Ca2+ concentration, cyclic AMP content and aggregating response to ADP and collagen were determined. Platelet from group HC patients were characterized by reduced cyclic AMP content (evaluated in the presence and absence of a platelet phosphodiesterase inhibitor) and aggregating responses to ADP and collagen, increased content and decreased arachidonic, eicosapentaenoic and docosatetraenoic and pentaenoic content, resulting in a lowered polyunsaturated to saturated fatty ratio (P less than 0.001). In contrast, platelet cytosolic Ca2+ concentration, DPH steady-state anisotropy and cholesterol to phospholipid molar ratio were not significantly changed. This indicates that hypercholesterolemia is accompanied in hypertensive patients by marked changes in platelet fatty composition, cyclic AMP content and response to aggregating agents. These changes, which clearly differ from those induced by in vitro cholesterol loading, could reflect not only the balance between LDL and HDL stimulation but also an adaptation to hemodynamic perturbations.

Keyword: hyperlipedemia

Hypolipidemic and hepatoprotective effects of flax and pumpkin seed mixture rich in omega-3 and omega-6 fatty acids in hypercholesterolemic rats.

Flax and pumpkin seeds are a rich source of unsaturated fatty acids, antioxidants and fibers, known to have anti-atherogenic and hepatoprotective activities. These effects were evaluated in Wistar rats fed with 1% cholesterol diet. The study was performed on 30 male rats divided into three groups: a control group (CD), CD-chol group fed diet with 1% cholesterol and MS-chol group fed diet enriched with flax and pumpkin seed mixture. In CD-chol group, total cholesterol TC, triacylglycerol TG in plasma and liver, plasma LDL-C, atherogenic index AI and LDL/HDL ratio significantly increased. In MS-chol group lipid parameters decreased significantly, plasma and liver fatty composition showed an increase of PUFAs (ALA and LA), and MUFAs (oleic and eicosaenoic ) and a decrease of SFA ( and stearic ). In plasma and liver of MS-chol group, malondialdehyde levels decreased and the efficiency of antioxidant defense system was improved compared to CD-chol group. Liver histological sections showed lipid storage in hepatocytes of CD-chol group and an improvement was noted in MS-chol group. Our results suggested that flax and pumpkin seed mixture had anti-atherogenic and hepatoprotective effects which were probably mediated by unsaturated fatty acids present in seed mixture.

Keyword: hyperlipedemia

Suppression of hepatocyte nuclear factor-4alpha by acyl-CoA thioesters of hypolipidemic peroxisome proliferators.

Hepatocyte nuclear factor-4alpha (HNF-4alpha) modulates the expression of liver-specific genes that control the production (e.g. apolipoprotein [apo] A-I and apo B) and clearance (e.g. apo C-III) of plasma lipoproteins. We reported that the CoA thioesters of amphipathic carboxylic hypolipidemic drugs (e.g. clofibric analogues currently used for treating in humans and substituted long-chain dicarboxylic acids) were formed in vivo, bound to HNF-4alpha, inhibited its transcriptional activity, and suppressed the expression of HNF-4alpha-responsive genes. Hypolipidemic PPARalpha (peroxisome proliferator-activated receptor alpha) activators that were not endogenously thioesterified into their respective acyl-CoAs were shown to be effective in rats but not in humans, implying that the hypolipidemic activity transduced by PPARalpha in rats was PPARalpha-independent in humans. The suppressed acyl-CoA synthase of PPARalpha knockout mice left unresolved the contribution made by the acyl-CoA/HNF-4alpha pathway to the hypolipidemic effect of PPARalpha agonists in rodents. Hence, suppression of HNF-4alpha activity by the CoA thioesters of hypolipidemic "peroxisome proliferators" may account for their hypolipidemic activity independently of PPARalpha activation by their respective free carboxylates. The hypolipidemic activity of peroxisome proliferators is mediated in rats and humans by the PPARalpha and HNF-4alpha pathways, respectively.

Keyword: hyperlipedemia

Ameliorative potential of S-allylcysteine: effect on lipid profile and changes in tissue fatty composition in experimental diabetes.

is an associated complication of diabetes mellitus. The association of hyperglycemia with an alteration of lipid parameters presents a major risk for cardiovascular complications in diabetes. The present study was designed to examine the antihyperlipidemic effect of S-allylcysteine (SAC) in STZ induced diabetic rats. The levels of blood glucose, cholesterol (TC), triglycerides (TG), free fatty acids, phospholipids and fatty composition were estimated in the liver and kidneys of control and experimental groups of rats. Oral administration of SAC at a dose of 150 mg/kg bodyweight per day to STZ-induced diabetic rats for a period of 45 days resulted in a significant reduction in fasting blood glucose, TC, TG, free fatty acids, phospholipids, LDL-C, VLDL-C and elevation of HDL-C in comparison with diabetic control group. Oral administration of SAC to diabetic rats also decreased the concentrations of fatty acids, viz., , stearic (16:1), and oleic (18:1), whereas linolenic (18:3) and arachidonic (20:4) were elevated. The antihyperlipidemic effect of SAC was compared with glyclazide; a well-known antihyperglycemic drug. The result of the present study indicates that SAC showed an antihyperlipidemic effect in addition to its antidiabetic effect in experimental diabetes.Copyright © 2010 Elsevier GmbH. All rights reserved.

Keyword: hyperlipedemia

[Fatty composition of triglycerides in inflammatory liver diseases in diabetics with and without hyperlipoproteinemia].

In 228 patients with diabetes mellitus (130 diabetics without and 98 diabetics with hyperlipoproteinaemia) percutaneous liver punctures after Menghini as well as biopsies of the subcutaneous fatty tissue were carried out. From the biopsy specimens and from serum the fatty pattern of triglycerides was estimated. In 87 patients with chronic aggressive hepatitis and 37 patients with fibrosis no differences in the fatty composition could be found. According to the present findings there was no evidence of alterations in the supply of individual fatty acids caused by chronic hepatitis per se. However, it is to be taken into consideration that simultaneous liver steatosis can provoke marked changes in the fatty pattern of liver triglycerides. The best reference seems to be the size of the fat droplet in the hepatocytes. Its rise is associated with an increase of and oleic , whereas the percentage of arachidonic and eicosapentaenoic is decreased. It must be clarified by further studies, whether this reveals a general pathophysiological phenomenon or is restricted to diabetic subjects.

Keyword: hyperlipedemia

Modulation of human postprandial lipemia by changing ratios of polyunsaturated to saturated (P/S) fatty content of blended dietary fats: a cross-over design with repeated measures.

Postprandial lipemia (PL) contributes to coronary artery disease. The fatty composition of dietary fats is potentially a modifiable factor in modulating PL response.This human postprandial study evaluated 3 edible fat blends with differing polyunsaturated to saturated fatty acids (P/S) ratios (POL\u2009=\u20090.27, AHA\u2009=\u20091.00, PCAN\u2009=\u20091.32). A cross-over design included mildly hypercholestrolemic subjects (9 men and 6 women) preconditioned on test diets fats at 31% energy for 7 days prior to the postprandial challenge on the 8th day with 50 g test fat. Plasma lipids and lipoproteins were monitored at 0, 1.5, 3.5, 5.5 and 7 hr.Plasma triacylglycerol (TAG) concentrations in response to POL, AHA or PCAN meals were not significant for time x test meal interactions (P\u2009>\u20090.05) despite an observed trend (POL\u2009>\u2009AHA\u2009>\u2009PCAN). TAG area-under-the-curve (AUC) increased by 22.58% after POL and 7.63% after PCAN compared to AHA treatments (P\u2009>\u20090.05). Plasma total cholesterol (TC) response was not significant between meals (P\u2009>\u20090.05). Varying P/S ratios of test meals significantly altered prandial high density lipoprotein-cholesterol (HDL-C) concentrations (P\u2009<\u20090.001) which increased with decreasing P/S ratio (POL\u2009>\u2009AHA\u2009>\u2009PCAN). Paired comparisons was significant between POL vs PCAN (P\u2009=\u20090.009) but not with AHA or between AHA vs PCAN (P\u2009>\u20090.05). A significantly higher HDL-C AUC for POL vs AHA (P\u2009=\u20090.015) and PCAN (P\u2009=\u20090.001) was observed. HDL-C AUC increased for POL by 25.38% and 16.0% compared to PCAN and AHA respectively. Plasma low density lipoprotein-cholesterol (LDL-C) concentrations was significant (P\u2009=\u20090.005) between meals and significantly lowest after POL meal compared to PCAN (P\u2009=\u20090.004) and AHA (P\u2009>\u20090.05) but not between AHA vs PCAN (P\u2009>\u20090.05). AUC for LDL-C was not significant between diets (P\u2009>\u20090.05). (C16:0), oleic (C18:1), linoleic (C18:2) and linolenic (C18:3) acids in TAGs and cholesteryl esters were significantly modulated by meal source (P\u2009<\u20090.05).P/S ratio of dietary fats significantly affected prandial HDL-C levels without affecting lipemia.

Keyword: hyperlipedemia

Effects of the individual saturated fatty acids on serum lipids and lipoprotein concentrations.

A mixture of dietary saturated fatty acids raises the serum total cholesterol concentrations compared with a diet of isoenergetic amounts of carbohydrates. Saturated fatty acids are not all equally hypercholesterolemic: stearic (18:0) and saturated fatty acids with < 12 carbon atoms are thought not to raise serum cholesterol concentrations. This suggest that the cholesterol-raising properties of saturated fatty acids should be attributed solely to lauric (12:0), myristic (14:0), and (16:0). These three saturated fatty acids, however, may have different effects on serum total-cholesterol concentrations as well. Results from controlled dietary experiments suggest that lauric (12:0) is less, and myristic (14:0) probably more, hypercholesterolemic than (16:0). Effects of the different saturated fatty acids on the distribution of cholesterol over the various lipoproteins are largely unknown, but it is suggested that stearic lowers HDL cholesterol concentrations relative to other saturated fatty acids. At present, however, too many questions are unanswered to quantitate these differences.

Keyword: hyperlipedemia

Altered plasma lysophosphatidylcholines and amides in non-obese and non-diabetic subjects with borderline-to-moderate hypertriglyceridemia: a case-control study.

Hypertriglyceridemia (HTG) is a risk factor for atherosclerotic cardiovascular disease (CVD). We investigated alterations in plasma metabolites associated with borderline-to-moderate HTG (triglycerides (TG) 150-500 mg/dL). Using UPLC-LTQ-Orbitrap mass spectrometry analysis, the metabolomics profiles of 111 non-diabetic and non-obese individuals with borderline-to-moderate HTG were compared with those of 111 age- and sex-matched controls with normotriglyceridemia (NTG, TG <150 mg/dL). When compared to the NTG control group, the HTG group exhibited higher plasma levels of lysophosphatidylcholines (lysoPCs), including C14:0 (q = 0.001) and C16:0 (q = 1.8E-05), and several amides, including N-ethyldodecanamide (q = 2.9E-05), N-propyldodecanamide (q = 3.5E-05), palmitoleamide (q = 2.9E-06), and amide (q = 0.019). The metabolomic profiles of the HTG group also exhibited lower plasma levels of cis-4-octenedioic (q<1.0E-9) and docosanamide (q = 0.002) compared with those of the NTG controls. LysoPC 16:0 and palmitoleamide emerged as the primary metabolites able to discriminate the HTG group from the NTG group in a partial least-squares discriminant analysis and were positively associated with the fasting triglyceride levels. We identified alterations in lysoPCs, amides, and cis-4-octenedioic among non-diabetic and non-obese individuals with borderline-to-moderate HTG. These results provide novel insights into the metabolic alterations that occur in the early metabolic stages of HTG. This information may facilitate the design of early interventions to prevent disease progression.

Keyword: hyperlipedemia

Blunted lipolysis and fatty oxidation during moderate exercise in HIV-infected subjects taking HAART.

The protease inhibitor (PI) ritonavir (RTV) has been associated with elevated resting lipolytic rate, , and insulin resistance/glucose intolerance. The purpose of this study was to examine relationships between lipolysis and fatty (FA) oxidation during rest, moderate exercise and recovery, and measures of insulin sensitivity/glucose tolerance and fat redistribution in HIV-positive subjects taking RTV (n=12), HAART but no PI (n=10), and HIV-seronegative controls (n=10). Stable isotope tracers [1-(13)C]palmitate and [1,1,2,3,3-(2)H5]glycerol were continuously infused with blood and breath collection during 1-h rest, 70-min submaximal exercise (50% VO2 peak), and 1-h recovery. Body composition was evaluated using DEXA, MRI, and MRS, and 2-h oral glucose tolerance tests with insulin monitoring were used to evaluate glucose tolerance and insulin resistance. Lipolytic and FA oxidation rates were similar during rest and recovery in all groups; however, they were lower during moderate exercise in both HIV-infected groups [glycerol Ra: HIV+RTV 5.1+/-1.2 vs. HIV+no PI 5.9+/-2.8 vs. Control 7.4+/-2.2 micromol.kg fat-free mass (FFM)-1.min-1; palmitate oxidation: HIV+RTV 1.6+/-0.8 vs. HIV+no PI 1.6+/-0.8 vs. Control 2.5+/-1.7 micromol.kg FFM.min, P<0.01]. Fasting and orally-challenged glucose and insulin values were similar among groups. Lipolytic and FA oxidation rates were blunted during moderate exercise in HIV-positive subjects taking HAART. Lower FA oxidation during exercise was primarily due to impaired plasma FA oxidation, with a minor contribution from lower nonplasma FA oxidation. Regional differences in adipose tissue lipolysis during rest and moderate exercise may be important in HIV and warrant further study.

Keyword: hyperlipedemia

[Effects of various modes of therapy on the fatty spectrum in hyperlipoproteinemia].

Gas-chromatographic investigations of the fatty pattern of the fractions of serum triglyceride and serum cholesterol ester revealed characteristic changes under different therapeutic measures in altogether 120 patients with primary hyperlipoproteinaemia of the types IIa, IIb and IV. Already under the treatment with basal metabolic diet, more clearly by medication with clofibrin , in the hyperlipoproteinaemia type IV the decrease of the , the palmitoleic , the stearic and oleic and the increase of linoleic , linoleneic , arachidonic and eikopentaenic in the triglyceride fraction developed. These fatty acids showed the same behaviour under the therapy with derivatives of nicotinic acids in the cholesterol ester fraction in the hyperlipoproteinaemia of type IIa. The combined treatment of the type IIb resulted in the same tendencies, however not so pronounced, of the spectrum in the cholesterol esters as well as of the fatty acids esterified in the triglycerides.

Keyword: hyperlipedemia

Palmitate-induced inflammatory pathways in human adipose microvascular endothelial cells promote monocyte adhesion and impair insulin transcytosis.

Obesity is associated with inflammation and immune cell recruitment to adipose tissue, muscle and intima of atherosclerotic blood vessels. Obesity and are also associated with tissue insulin resistance and can compromise insulin delivery to muscle. The muscle/fat microvascular endothelium mediates insulin delivery and facilitates monocyte transmigration, yet its contribution to the consequences of is poorly understood. Using primary endothelial cells from human adipose tissue microvasculature (HAMEC), we investigated the effects of physiological levels of fatty acids on endothelial inflammation and function. Expression of cytokines and adhesion molecules was measured by RT-qPCR. Signaling pathways were evaluated by pharmacological manipulation and immunoblotting. Surface expression of adhesion molecules was determined by immunohistochemistry. THP1 monocyte interaction with HAMEC was measured by cell adhesion and migration across transwells. Insulin transcytosis was measured by total internal reflection fluorescence microscopy. Palmitate, but not palmitoleate, elevated the expression of IL-6, IL-8, TLR2 (Toll-like receptor 2), and intercellular adhesion molecule 1 (ICAM-1). HAMEC had markedly low fatty uptake and oxidation, and CD36 inhibition did not reverse the palmitate-induced expression of adhesion molecules, suggesting that inflammation did not arise from palmitate uptake/metabolism. Instead, inhibition of TLR4 to NF-κB signaling blunted palmitate-induced ICAM-1 expression. Importantly, palmitate-induced surface expression of ICAM-1 promoted monocyte binding and transmigration. Conversely, palmitate reduced insulin transcytosis, an effect reversed by TLR4 inhibition. In summary, palmitate activates inflammatory pathways in primary microvascular endothelial cells, impairing insulin transport and increasing monocyte transmigration. This behavior may contribute in vivo to reduced tissue insulin action and enhanced tissue infiltration by immune cells.Copyright © 2015 the American Physiological Society.

Keyword: hyperlipedemia

Lipid-stimulated somatostatin secretion in rainbow trout,Oncorhynchus mykiss.

Previous work has shown that somatostatins (SS) affect teleost lipid metabolism indirectly by inhibition of insulin (INS) and directly by stimulation of hepatic lipolysis. In the present study, rainbow trout (Oncorhynchus mykiss) were used to characterize further the lipid-SS relationship by evaluating how lipid, contributes to SS secretion bothin vivo andin vitro. In vivo was induced for up to 3 h by short-term (2 min) infusion of a triacylglycerol (TG)-rich lipid emulsion (20% Intralipid(®)). Plasma total lipid concentration increased 118 and 155% over control levels 1 h and 3 h, respectively, after infusion; much of this increase was due to elevated plasma fatty acids (FA), which increased 39 and 520%, respectively, over the same time-frame. The hyperlipidemic pattern was attended by a significant increase in the plasma concentration of SS. The specific effects of fatty acids were evaluated on isolated Brockmann bodies. and oleic stimulated SS release 378 and 82%, respectively, over baseline levels. These results indicate that lipids, and in particular fatty acids, modulate SS secretion in rainbow trout.

Keyword: hyperlipedemia

The fatty liver dystrophy mutant mouse: microvesicular steatosis associated with altered expression levels of peroxisome proliferator-regulated proteins.

Fatty liver dystrophy ( fld) is an autosomal recessive mutation in mice characterized by hypertriglyceridemia and fatty liver during neonatal development. The fatty liver in fld/fld mice spontaneously resolves between the age of 14-18 days, at which point the animals develop a neuropathy associated with abnormal myelin formation in peripheral nerve. We have investigated the morphological and biochemical alterations that occur in the fatty liver of neonatal fld/fld mice. Studies at the light and electron microscopic level demonstrated the accumulation of lipid droplets and hypertrophic parenchymal cells in fld neonates, with no apparent liver pathology after resolution of the fatty liver. To better characterize the biochemical basis for the development of fatty liver in fld mice, we compared protein expression patterns in the fatty liver of fld mice and in the liver of phenotypically normal (wild-type) littermates using quantitative two-dimensional gel electrophoresis. We detected 24 proteins with significantly altered expression levels (P < 0.001) in the fld fatty liver, 15 of which are proteins that are altered in abundance by peroxisome proliferating chemicals. As these compounds characteristically elicit changes in the expression of mitochondrial and peroxisomal enzymes involved in fatty oxidation, we quantitated rates of fatty oxidation in hepatocytes isolated from fld and wild-type mice. These studies revealed that hepatic fatty oxidation in fld neonates is reduced by 60% compared to wild-type littermates. In hepatocytes from adult fld mice that no longer exhibit a fatty liver, oxidation rates were similar to those in hepatocytes from age-matched wild-type mice. These findings indicate that altered expression of proteins involved in fatty oxidation is associated with triglyceride accumulation in the fld fatty liver.

Keyword: hyperlipedemia

[Clinico-experimental long-term study on the combined effect of clofibrin and nicotinic on abnormal fatty patterns in lipid metabolism disorders in the aged].

Seventy-one patients (forty-two males and twenty-nine females aged forty-five to seventy-six) with disturbances of fat metabolism were treated for three years with clofibric and nicotinic derivatives. Regular gas chromatographic analyses of the composition of cholesterol ester and triglyceride fatty acids in the serum showed an increase of linoleic, linolenic, arachidonic, and eicosane-pentaenoic acids and a decrease of , palmitoleic, stearic, oleic, and eicosanetrienic acids during treatment. These changes were far more strongly marked than under monotherapy. Possible causes of the observed changes include selective competitive inhibition of unesterified serum fatty acids, inhibition of lipolysis, effects upon hepatogenic fatty metabolism and LCAT, as well as better utilization of alimentary polyunsaturated fatty acids as a result of combination treatment. The increase of polyunsaturated fatty acids and the decrease of saturated and monounsaturated fatty acids with their reciprocal relations to the prostaglandin metabolism may be considered a positive vasoprotective effect which is of particular importance in middle and old age.

Keyword: hyperlipedemia

Peroxisome proliferators as adjuvants for the reverse-electron-transport therapy of obesity: an explanation for the large increase in metabolic rate of MEDICA 16-treated rats.

The efficacy of reverse-electron-transport therapy of obesity should be promoted by agents which up-regulate hepatocyte enzymes that are potentially rate-limiting for mitochondrial fatty oxidation and electron shuttles. Peroxisome proliferator drugs, including the fibrates used to treat , may be useful in this regard, as they induce malic enzyme, the mitochondrial glycerol-3-phosphate dehydrogenase, and carnitine palmitoyl transferase I in rodent hepatocytes. An agent of this class, MEDICA 16, has the additional property of potently inhibiting both citrate lyase and acetyl-CoA carboxylase. As a result, methyl-substituted diacarboxylic acids (MEDICA) 16 can be expected to disinhibit hepatic fatty oxidation while up-regulating electron shuttle mechanisms, and thus should stimulate reverse electron transport. This may explain the remarkable 40% increase in basal metabolic rate observed in normal rats ingesting MEDICA 16--an effect not associated with any compensatory increase in food intake. Relative to controls, the MEDICA 16-treated rats achieved a 50% reduction in body fat and a modest increase in lean mass, such that weight and growth were not changed. In other rodent strains, MEDICA 16 has prevented obesity diabetes and atherogenesis. However, whether MEDICA 16 and other peroxisome proliferator drugs will have clinical utility in reverse-electron-transport therapy may hinge on their ability to induce key enzymes in human hepatocytes; cell culture studies to evaluate this are required.

Keyword: hyperlipedemia

An anti-inflammatory chalcone derivative prevents heart and kidney from -induced injuries by attenuating inflammation.

Obesity is a growing pandemic in both developed and developing countries. Lipid overload in obesity generates a chronic, low-grade inflammation state. Increased inflammation in heart and renal tissues has been shown to promote the progression of heart and renal damage in obesity. Previously, we found that a novel chalcone derivative, L6H21, inhibited lipopolysaccharide-induced inflammatory response. In the present study, we investigated the effects of L6H21 on inflammatory responses in culture and in animal models of lipid overload. We utilized (PA) challenging in mouse peritoneal macrophages and apolipoprotein E knockout (ApoE) mice fed a high fat diet (HFD) to study whether L6H21 mitigates the inflammatory response. Our studies show that L6H21 significantly reduced PA-induced expression of inflammatory cytokines in macrophages by inhibiting mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NFκB) signaling pathways. L6H21 also reduced fibrosis in the kidney and heart tissues, and indices of inflammatory response in the ApoE mice fed a HFD. These effects in vivo were also associated with inhibition of MAPK and NFκB signaling by L6H21. These findings strongly suggest that L6H21 may be a potential agent for high fat diet-induced injuries in heart and kidney.Copyright © 2017. Published by Elsevier Inc.

Keyword: hyperlipedemia

Chalcones suppress fatty -induced lipid accumulation through a LKB1/AMPK signaling pathway in HepG2 cells.

Excessive lipid accumulation in the liver has been proposed to cause , diabetes and fatty liver disease. 4-Hydroxyderricin (4HD), xanthoangelol (XAG), cardamonin (CAR) and flavokawain B (FKB) are chalcones that have exhibited various biological effects against obesity, inflammation, and diabetes; however, little is known about the inhibitory effects of these chalcones on fatty liver disease. In the present study, we investigated the ability of 4HD, XAG, CAR, and FKB to reduce lipid accumulation in hepatocytes. When HepG2 cells were treated with a mixture of fatty acids (FAs; : oleic = 1 : 2 ratio), significant lipid accumulation was observed. Under the same experimental conditions, addition of chalcones at 5 μM significantly suppressed the FA-induced lipid accumulation. We found that the expression of sterol regulatory element-binding protein-1 (SREBP-1), a key molecule involved in lipogenesis, was decreased in these chalcone-treated cells. We also found that these chalcones increased the expression of peroxisome proliferator-activated receptor α (PPARα), which is involved in FA oxidation. Moreover, these chalcones increased phosphorylation of AMP-activated protein kinase (AMPK) and liver kinase B1 (LKB1), upstream regulators of SREBP-1 and PPARα. We confirmed that an AMPK inhibitor, compound C, reversed chalcone-induced changes in SREBP-1 and PPARα expression in the HepG2 cells. Collectively, we found that 4HD, XAG, CAR, and XAG attenuated lipid accumulation through activation of the LKB1/AMPK signaling pathway in HepG2 cells.

Keyword: hyperlipedemia

Familial hypertriglyceridemia and the regulation of lipid synthesis by insulin and triiodothyronine in human cell cultures.

Keyword: hyperlipedemia

Relationship between fatty acids of adipose tissue and plasma cholesterol.

The fatty composition of adipose tissue from normal and hypercholesterolemic men and women was studied. We found a difference in the fatty composition between both sexes. A decrease in linoleic was found in hypercholesterolemic men, while there was an increase in and totally saturated acids in women. A decrease in total monounsaturated and in monounsaturated/saturated ratio was also found in women.

Keyword: hyperlipedemia

PTEN phosphorylation and nuclear export mediate free fatty -induced oxidative stress.

Oxidative stress induced by free fatty acids (FFA) contributes to metabolic syndrome-associated development of cardiovascular diseases, yet molecular mechanisms remain poorly understood. This study aimed at establishing whether phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and its subcellular location play a role in FFA-induced endothelial oxidative stress.Exposing human endothelial cells (ECs) with FFA activated mammalian target of rapamycin (mTOR)/S6K pathway, and upon activation, S6K directly phosphorylated PTEN at S380. Phosphorylation of PTEN increased its interaction with its deubiquitinase USP7 in the nucleus, leading to PTEN deubiquitination and nuclear export. The reduction of PTEN in the nucleus, in turn, decreased p53 acetylation and transcription, reduced the expression of the p53 target gene glutathione peroxidase-1 (GPX1), resulting in reactive oxygen species (ROS) accumulation and endothelial damage. Finally, C57BL/6J mice fed with high-fat atherogenic diet (HFAD) showed PTEN nuclear export, decreased p53 and GPX1 protein expressions, elevated levels of ROS, and significant lesions in aortas. Importantly, inhibition of mTOR or S6K effectively blocked these effects, suggesting that mTOR/S6K pathway mediates HFAD-induced oxidative stress and vascular damage via PTEN/p53/GPX1 inhibition in vivo.Our study demonstrated for the first time that S6K directly phosphorylated PTEN at S380 under high FFA conditions, and this phosphorylation mediated FFA-induced endothelial oxidative stress. Furthermore, we showed that S380 phosphorylation affected PTEN monoubiquitination and nuclear localization, providing the first example of coordinated regulation of PTEN nuclear localization via phosphorylation and ubiquitination.Our studies provide a novel mechanism by which causes vascular oxidative damage through the phosphorylation of PTEN, blocking of PTEN nuclear function, and inhibition of p53/GPX1 activity.

Keyword: hyperlipedemia

Lipidomics identification of metabolic biomarkers in chemically induced hypertriglyceridemic mice.

In this study, we aim to identify the potential biomarkers in hTG pathogenesis in schisandrin B-induced hTG mouse model. To investigate whether these identified biomarkers are only specific to schisandrin B-induced hTG mouse model, we also measured these biomarkers in a high fat diet (HFD)-induced hTG mouse model. We employed a LC/MS/MS-based lipidomic approach for the study. Mouse liver and serum metabolites were separated by reversed phase liquid chromatography. Metabolite candidates were identified by matching with marker retention times, isotope distribution patterns, and high-resolution MS/MS fragmentation patterns. Subsequently, target candidates were quantified by quantitative MS. In the schisandrin B-induced hTG mice, we found that the plasma fatty acids, diglyceroids, and phospholipids were significantly increased. and stearic were increased in the plasma; oleic , linoleic , linolenic , arachidonic , and docosahexaenoic were increased in both the plasma and the liver. Acetyl-CoA, malonyl-CoA, and succinyl-CoA were increased only in the liver. The changes in levels of these identified markers were also observed in HFD-induced hTG mouse model. The consistent results obtained from both hTG models not only suggest novel biomarkers in hTG pathogenesis, but they also provide insight into the underlying mechanism of the schisandrin B-induced hTG.

Keyword: hyperlipedemia

Plasma lipoproteins and the synthesis and turnover of plasma triglyceride in normal and genetically obese mice.

1. Lipoproteins in the plasma of mice were characterized by agarose-gel chromatography and polyacrylamide-gel electrophoresis: genetically obese (ob/ob) mice exhibited hyperlipoproteinaemia (compared with lean mice), largely owing to an increase in the concentration of cholesterol in high-density lipoprotein. Plasma concentrations of triglyceride and phospholipid were not markedly increased in genetically obese mice. 2. The formation of glycerolipids in liver and plasma was investigated with (14)C-labelled precursors. The synthesis of hepatic triglyceride and phospholipid from glucose or palmitate was enhanced in ob/ob mice, compared with lean mice. The rate of entry of triglyceride into plasma, calculated from the time-course of incorporation of (14)C from [(14)C]palmitate into plasma triglyceride, was increased in ob/ob mice (0.5mumol of fatty /min, compared with 0.2 in lean mice). 3. The removal from plasma of murine lipoprotein triglyceride-[(14)C]fatty was increased in ob/ob mice (half-time 2.2min, compared with 7.2min in lean mice). Similar results were obtained with an injected lipid emulsion (Intralipid). 4. From these measurements, estimates of the rates of turnover of plasma triglyceride in mice (fed on a mixed diet, female, 3 months old) are about 1.0mumol of fatty /min in ob/ob mice, and 0.25 in lean mice. 5. The major precursor of hepatic and plasma triglyceride in lean and ob/ob mice was calculated to be plasma free fatty . 6. These results are discussed, in connexion with the role of the liver in triglyceride metabolism in mice, especially in relation to genetic obesity.

Keyword: hyperlipedemia

Daytime restricted feeding modifies the daily regulation of fatty β-oxidation and the lipoprotein profile in rats.

Daytime restricted feeding (2 h of food access from 12.00 to 14.00 hours for 3 weeks) is an experimental protocol that modifies the relationship between metabolic networks and the circadian molecular clock. The precise anatomical locus that controls the biochemical and physiological adaptations to optimise nutrient use is unknown. We explored the changes in liver oxidative lipid handling, such as β-oxidation and its regulation, as well as adaptations in the lipoprotein profile. It was found that daytime restricted feeding promoted an elevation of circulating ketone bodies before mealtime, an altered hepatic daily rhythmicity of 14CO2 production from radioactive , and an up-regulation of the fatty oxidation activators, the α-subunit of AMP-activated protein kinase (AMPK), the deacetylase silent mating type information regulation homolog 1, and the transcriptional factor PPARγ-1α coactivator. An increased localisation of phosphorylated α-subunit of AMPK in the periportal hepatocytes was also observed. Liver hepatic lipase C, important for lipoprotein transformation, showed a change of daily phase with a peak at the time of food access. In serum, there was an increase of LDL, which was responsible for a net elevation of circulating cholesterol. We conclude that our results indicate an enhanced fasting response in the liver during daily synchronisation to food access, which involves altered metabolic and cellular control of fatty oxidation as well a significant elevation of serum LDL. These adaptations could be part of the metabolic input that underlies the expression of the food-entrained oscillator.

Keyword: hyperlipedemia

[THE EFFECT OF SATINS: ACTIVATION OF LIPOLYSIS AND ABSORPTION BY INSULIN-DEPENDED CELLS LIPOPROTEINS OF VERY LOW DENSITY, INCREASING OF BIO-AVAILABILITY OF POLYENOIC FATTY ACIDS AND DECREASING OF CHOLESTEROL OF LIPOPROTEINS OF LOW DENSITY].

The Russian cardiologic R&D production complex of Minzdrav of Russia, 121552 Moscow, Russia The statins are synthetic xenobiotics alien to animal cells. They are unlikely capable to manifest pleiotropic effect. It is feasible to evaluate effect of statins by stages: a) initially a specific inhibition of synthesis of cholesterol alcohol; b) further indirect activation of hydrolysis of triglycerides in lipoproteins of very low density; c) nonspecific activation of cells\' receptor absorption of and oleic lipoproteins of very low density and then d) linoleic and linolenic lipoproteins of low density with all polyenoic fatty acids. On balance, statins activate absorption ofpolyenoic fatty acids by cells. Just they manifest physiological, specific pleiotropic effect. The statins inhibit synthesis of pool of cholesterol alcohol-lipoproteins of very low density condensed between phosphatidylcholines in polar mono-layer phosphatidylcholines+cholesterol alcohol on surface oftriglycerides. The low permeability of mono-layer separates substrate-triglycerides in lipoproteins of very low density and post-heparin lipoprotein lipase in hydrophilic blood plasma. The higher is ratio cholesterol alcohol/phosphatidylcholines in mono-layer of lipoproteins of very low density the slower is lipolysis, formation of ligand lipoproteins of very low density and their absorption by cells under apoB-100-endocytosis. The statins normalize hyperlipemia by force of a) activation of absorption oflipoproteins of very low density by insulin-depended cells and b) activation of absorption of lipoproteins of low density by all cells, increasing of bio-availability of polyenoic fatty acids, activation of apoB-100-endocytosis. The limitation in food of content of saturated fatty and increasing of content of ω-3 polyenoic fatty acids improve "bio-availability" of polyenoic fatty acids and their absorption by cells and also decreases cholesterol alcohol/phosphatidylcholines and biological pleiotropic effect of essential polyenoic fatty acids. According our opinion, atherosclerosis is intracellular deficiency of polyenoic fatty acids. The value of cholesterol alcohol-lipoproteins of low density is equimolar to content of lipoproteins of low density in blood which under low bio-availability can\'t to absorb cells byforce of apoB-100-endocytosis.

Keyword: hyperlipedemia

Adipose tissue glyceride synthesis in patients with hyperapobetalipoproteinemia.

Adipose tissue was obtained at thoracotomy in five control patients with valvular heart disease, all of whom were free of coronary artery disease and all of whom were normolipidemic with normal low density lipoprotein apolipoprotein B levels, and eight patients with coronary artery disease, all of whom had hyperapobetalipoproteinemia. In both groups, the rates at which linoleic and were incorporated into diglyceride and triglyceride were determined in vitro. The data indicate that fatty incorporation into adipose tissue glycerides was twice as rapid in controls as in patients with hyperapobetalipoproteinemia. By contrast there was no difference between the groups in the rate of net lipolysis of adipocyte glyceride. The data at hand do not establish the mechanism responsible for the difference in synthesis between normal subjects and patients with hyperapobetalipoproteinemia, but this may explain the delayed chylomicron triglyceride clearance previously observed in the disorder.

Keyword: hyperlipedemia

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial lipid accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic lipid metabolism. Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 diabetes animal models. After streptozotocin (STZ) or high-fat diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, , myocardial hypertrophy, fibrosis, lipid accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and lipid accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing lipid accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

[Human ethanol-induced hyperlipoproteinemia (author\'s transl)].

Reversible hyperlipoproteinemia may be observed after ethanol loads in healthy man before any ethanol-induced disease is being established. Different pathogenetic ways to this acute ethanol-induced hyperlipoproteinemia have been investigated or postulated in recent years. Two main sites have appeared: changes in the metabolism of lipids and their precursors which depend from acutal oxidation of ethanol in the liver, and ethanol-induced activation of lipolysis in adipose tissue, transmitted by the sympathico-adrenal system. The changes in liver metabolism during ethanol oxidation have been well confirmed in many experiments, they nevertheless do not seem to lead to hyperlipoproteinemia in many experimental designs in animals and after drinkable amounts of ethanol in healthy man when lipolysis of adipose tissue is blocked and no food is ingested. After the intake of a fatty meal these triglycerides are becoming importance as a source of fatty acids. A possible increased de novo synthesis of may to a minor degree contribute to hypertriglyceridemia.

Keyword: hyperlipedemia

Effect of Euterpe oleracea Mart. (Açaí) Oil on Dyslipidemia Caused by Cocos nucifera L. Saturated Fat in Wistar Rats.

Dyslipidemia is caused by disturbances in lipid metabolism that lead to chronic elevations of serum lipids, especially low-density lipoprotein (LDL)-cholesterol and triglycerides, increasing the risk of metabolic syndrome, obesity, diabetes, atherogenic processes, and cardiovascular diseases. The oil from the fruits of Euterpe oleracea (OFEO) is rich in unsaturated fatty acids with potential for treating alterations in lipid metabolism. In this study, we aimed to investigate the effect of OFEO on induced by Cocos nucifera L. saturated fat (GSC) in Wistar rats. Chromatographic profile showed that unsaturated fatty acids account for 66.08% in OFEO, predominately oleic (54.30%), and saturated fatty acids ( 31.6%) account for 33.92%. GSC-induced dyslipidemia resulted in an increase in total cholesterol, LDL-cholesterol, triglycerides, glucose, and liver and abdominal fat, as well as atherogenic processes in the thoracic aorta. OFEO treatment did not reduce hypertriglyceridemia, but did reduce total cholesterol and LDL-cholesterol, thus contributing to the antiatherogenic action of OFEO. OFEO treatment inhibited the formation of atheromatous plaques in the vascular endothelium of the treated rats, as well as those who were treated with simvastatin. The results obtained suggest that OFEO has an antiatherogenic effect in a rat model of dyslipidemia.

Keyword: hyperlipedemia

Alterations of fatty metabolism in liver cells of hyperlipaemic old rats.

Keyword: hyperlipedemia

One-year effects of increasingly fat-restricted, carbohydrate-enriched diets on lipoprotein levels in free-living subjects.

Restriction of all dietary fat is a popular strategy for restricting saturated fat intake to lower LDL cholesterol. Some authorities advise the restriction of fat intake to the extreme of less than 10% of daily energy on the assumption that more fat restriction is better. The two studies described herein address questions relating to whether increasing fat restriction produces proportionally increasing benefit on cardiovascular risk factors in hyperlipidemic subjects. The first study is the Dietary Alternatives Study (DAS). The DAS was conducted in 531 male Boeing employees over a 2-year period. Subjects were defined as hypercholesterolemic (HC) or combined hyperlipidemic (CHL) based on age-specific 75th percentiles for plasma LDL-C and triglyceride levels. Hypothesis test analyses were performed at 1 year. HC subjects were randomized to diets taught to attain fat intakes of 30, 26, 22, and 18% (Diets levels 1-4, respectively). CHL subjects (slightly fewer in number) were randomized to Diets 1-3. After 1 year, subjects\' total fat intakes were 27, 26, 25, and 22% of energy (en%), resulting in saturated fat intakes of 8, 7, 7, and 6%, respectively. In HC subjects the greatest LDL-C decrease was with Diet 2 (mean of 13.4%) and in CHL subjects with Diet 1 (7.0%). Surprisingly, plasma triglyceride concentrations rose in HC subjects 20% and 40% above baseline on Diets 3 and 4, respectively, with reciprocal reductions in HDL cholesterol of 2.5% and 3%, respectively. Furthermore, apo B reductions were attenuated below Diet 2 in HC subjects and Diet 1 in CHL subjects, and no further reductions were seen in plasma glucose and insulin concentrations, blood pressure, or body weight. Measurements of plasma total fatty composition showed a slight increase in plasma palmitate, whereas stearate decreased slightly, supporting the idea that de novo synthesis of was increased in the chronic high-carbohydrate feeding condition. The second study asked if the most effective diet in HC subjects, Diet 2, has an equivalent effect in women and men. To answer this question, men and women Boeing employees were taught the closely similar National Cholesterol Education Program (NCEP) Step II diet. After 6 and 12 months, equivalent reductions in LDL cholesterol were observed in women compared with men. HDL cholesterol levels in men were unchanged from baseline at 6 and 12 months, but were reduced 8% in HC women, with accompanying decreases of 18% in HDL2-cholesterol and 5% in apoprotein A-I (all P < 0.01). These data indicate that intakes of fat below about 25 en% and carbohydrate intake above approximately 60 en% yield no further LDL-C lowering in HC and CHL male subjects and can be counterproductive to triglyceride, HDL-C, and apo B levels. This lack of benefit appears to be explained by an enhanced endogenous synthesis of , which negates the benefit of further saturated fat restriction. The HDL-C decrease in HC women may have a similar cause and points to an underlying male-female difference. Alternative dietary approaches to limit saturated fat intake deserve intensive study.

Keyword: hyperlipedemia

Changes in the metabolism of fatty acids in adipose tissue in obese patients with primary hypertriacylglycerolemia.

Changes in the release and esterification of free fatty acids (FFA) in adipose tissue were looked for as a cause of moderate primary hypertriacylglycerolemia (HTG) in five obese subjects. Comparison was made with six obese normolipidemic subjects. The two groups were matched for body weight, tolerance of intravenous glucose, fat cell size, fasting levels of serum immunoreactive insulin, and serum insulin response to an intravenous glucose load. Subcutaneous adipose tissue was incubated in vitro with [1-14C] for 30, 60, and 120 min. There was a significant, twofold increase in the rate of FFA mobilization, but no change of glycerol release in HTG patients. The adipose tissue levels of mono- and diacylglycerols were similar in the two groups of subjects and did not change during incubation. Re-esterification of FFA, calculated from the net changes in medium and in tissue FFA and glycerol release, was lower in HTG patients than in the controls (3 and 12 mumol/10(7) cells/hr, respectively; P less than 0.025). In adipose tissue of HTG patients, the amount of radioactive fatty acids incorporated into triacylglycerols (TG) was 50% lower (P less than 0.025), whereas that incorporated into tissue FFA was three times higher (P less than 0.01) when compared with control patients. It is concluded that, in adipose tissue of obese patients with primary hypertriacylglycerolemia, the esterification of free fatty acids to triacylglycerol is decreased. As a consequence, free fatty acids are mobilized at an increased rate.

Keyword: hyperlipedemia

GPR40 protein levels are crucial to the regulation of stimulated hormone secretion in pancreatic islets. Lessons from spontaneous obesity-prone and non-obese type 2 diabetes in rats.

The role of islet GPR40 protein in the pathogenesis of diabetes is unclear. We explored the influence of GPR40 protein levels on hormone secretion in islets from two rat models of spontaneous type 2 diabetes displaying either or hyperglycaemia. GPR40 expression was analysed by confocal microscopy, Western blot and qPCR in islets from preobese Zucker (fa/fa) rats, diabetic Goto-Kakizaki (GK) rats, and controls. Confocal microscopy of control islets showed expression of GPR40 protein in insulin, glucagon and somatostatin cells. GPR40 expression was strongly increased in islets of hyperlipidaemic fa/fa rats and coincided with a concentration-related increase in palmitate-induced release of insulin and glucagon and its inhibition of somatostatin release. Conversely, hyperglycaemic GK islets displayed an extremely faint expression of GPR40 as did high-glucose-cultured control islets. This was reflected in abolished palmitate-induced hormone response in GK islets and high-glucose-cultured control islets. The palmitate antagonist rosiglitazone promoted reappearance of GPR40 in high-glucose-cultured islets and served as partial agonist in glucose-stimulated insulin release. GPR40 protein is abundantly expressed in pancreatic islets and modulates stimulated hormone secretion. Mild in obesity-prone diabetes creates increased GPR40 expression and increased risk for an exaggerated palmitate-induced insulin response and lipotoxicity, a metabolic situation suitable for GPR40 antagonist treatment. Chronic hyperglycaemia creates abrogated GPR40 expression and downregulated insulin release, a metabolic situation suitable for GPR40 agonist treatment to avoid glucotoxicity. GPR40 protein is interactively modulated by both free fatty acids and glucose and is a promising target for pharmacotherapy in different variants of type 2 diabetes.Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Keyword: hyperlipedemia

Effects of diet and simvastatin on fatty composition in hypercholesterolemic men: a randomized controlled trial.

To explore the separate and combined effects of simvastatin and a low-saturated diet rich in alpha-linolenic on serum fatty acids.120 hypercholesterolemic men were randomly allocated to a habitual diet or dietary treatment group and to receive, in random order, simvastatin 20 mg/d or placebo, each for 12 weeks, in a double-blind manner. Dietary treatment decreased proportions from total fatty acids of (C16:0) by 3.3% (P<0.05), stearic (C18:0) by 3.7% (P<0.05) and increased proportions of oleic (C18:1n-9) by 4.2% (P<0.01), and alpha-linolenic (C18:3n-3) by 29.8% (P<0.001). Simvastatin decreased proportions from total fatty acids of by 2.0% (P<0.01), linoleic (C18:2n-6) by 5.3% (P<0.001), and alpha-linolenic by 6.8% (P<0.05), and increased proportions of gamma-linolenic (C18:3n-6) by 11.1% (P<0.001), dihomo-gamma-linolenic (C20:3n-6) by 4.2% (P<0.01), arachidonic (C20:4n-6) by 14.2% (P<0.001), and the sum of long-chain polyunsaturated fatty acids (C20-22) by 9.0% (P<0.001). Simvastatin increased ratios of stearic to , gamma-linolenic to linoleic, and arachidonic to dihomo-gamma-linolenic by 7.6%, 17.0%, and 10.0% (P<0.001 for all), respectively, suggesting increased fatty elongase and Delta6- and Delta5-desaturase enzyme activities.Increased formation of long-chain polyunsaturated fatty acids and their metabolites may contribute a substantial part of the pleiotropic effects of simvastatin.

Keyword: hyperlipedemia

Glucose and fatty acids synergize to promote B-cell apoptosis through activation of glycogen synthase kinase 3β independent of JNK activation.

The combination of elevated glucose and free-fatty acids (FFA), prevalent in diabetes, has been suggested to be a major contributor to pancreatic β-cell death. This study examines the synergistic effects of glucose and FFA on β-cell apoptosis and the molecular mechanisms involved. Mouse insulinoma cells and primary islets were treated with palmitate at increasing glucose and effects on apoptosis, endoplasmic reticulum (ER) stress and insulin receptor substrate (IRS) signaling were examined.Increasing glucose (5-25 mM) with palmitate (400 µM) had synergistic effects on apoptosis. Jun NH2-terminal kinase (JNK) activation peaked at the lowest glucose concentration, in contrast to a progressive reduction in IRS2 protein and impairment of insulin receptor substrate signaling. A synergistic effect was observed on activation of ER stress markers, along with recruitment of SREBP1 to the nucleus. These findings were confirmed in primary islets. The above effects associated with an increase in glycogen synthase kinase 3β (Gsk3β) activity and were reversed along with apoptosis by an adenovirus expressing a kinase dead Gsk3β.Glucose in the presence of FFA results in synergistic effects on ER stress, impaired insulin receptor substrate signaling and Gsk3β activation. The data support the importance of controlling both hyperglycemia and in the management of Type 2 diabetes, and identify pancreatic islet β-cell Gsk3β as a potential therapeutic target.

Keyword: hyperlipedemia

Hypertriglyceridemia aggravates ER stress and pathogenesis of acute pancreatitis.

Endoplasmic reticulum (ER) stress and hypertriglyceridemia (HTG) have been implicated in acute pancreatitis (AP).For cellular model, rat exocrine acinar cells were preincubated with (0.05 or 0.1 mmol/L, 3 h) and stimulated with a cholecystokinin analog, CCK-8 (100 pmol/L, 30 min). For animal model, rats fed a high-fat diet to cause HTG and AP was induced by injection of caerulein (20 μg/kg). Injury to pancreatic cells was estimated by measuring amylase secretion, intracellular calcium concentration, apoptosis and histological changes. Expression of genes involved in ER stress-induced unfolded protein response (UPR) was monitored by RT-PCR and immunohistology.In CCK-8 stimulated rat acinar cells, preincubation with PA caused an increased secretion of amylase, a higher and prolonged accumulation of intracellular calcium and increased apoptosis. Rats on high-fat diet had significantly elevated serum triglyceride levels. Induction of AP led to increased apoptosis in pancreatic tissue on high-fat diet than controls. For favoring HTG, expression of UPR components, GRP78/Bip, XBP-1, GADD153/CHOP and caspase-12 was upregulated.Levels of markers of AP pathogenesis and components of UPR were elevated in the presence of excess fatty acids in pancreatic acinar cells. HTG appears to aggravate ER-stress and pathogenesis of AP.

Keyword: hyperlipedemia

Behenic is a cholesterol-raising saturated fatty in humans.

Dietary behenic (22:0) is poorly absorbed. Because of its low bioavailability compared with other fatty acids and because of its very long chain length, the effect of dietary behenic (behenate) on serum lipid concentrations in humans is assumed to be neutral.The objective was to establish the cholesterol-raising potential of behenic by comparing the effects on lipid and lipoprotein concentrations of a specially formulated fat enriched with behenic with those of palm oil (rich in ; 16:0) and high-oleic sunflower oil (rich in cis oleic ; 18:1).In a randomized, crossover, metabolic-ward study, 7 mildly hypercholesterolemic men were fed 3 natural-food diets supplemented with behenate oil, palm oil, or high-oleic sunflower oil. Mean serum lipid and lipoprotein concentrations and plasma triacylglycerol fatty composition were determined from fasting blood drawn during the final 4 d of each 3-wk diet period.Behenate oil produced mean concentrations of total cholesterol (5.87+/-0.8 mmol/L) and LDL cholesterol (4.40+/-0.8 mmol/L) not significantly different from those produced by palm oil (5.84+/-0.7 and 4.42+/-0.7 mmol/L, respectively) but significantly higher than those produced by high-oleic sunflower oil (5.12+/-0.5 and 3.70+/-0.6 mmol/L, respectively). There were no significant differences in triacylglycerol or HDL-cholesterol concentrations.Despite its low bioavailability compared with oleic , behenic is a cholesterol-raising fatty in humans and is therefore not a suitable substitute for in manufactured triacylglycerols.

Keyword: hyperlipedemia

Hyperglycemia magnifies Schwann cell dysfunction and cell death triggered by PA-induced lipotoxicity.

Lipid overload resulting in lipotoxicity is prominent in a number of chronic diseases and has been associated with cellular dysfunction and cell death. This study characterizes -induced lipotoxicity (PA-LTx) in Schwann cell cultures grown in normal and high glucose concentrations. The study shows for the first time that Schwann cell (SC) cultures exposed to elevated levels of PA exhibit a dose- and time-dependent loss in cell viability. Hoescht and Annexin V/7AAD staining confirmed cell death through apoptosis and the lipotoxic effect was more dramatic in SC cultures grown under high glucose conditions. The first indication of cellular dysfunction in treated SC cultures was a decrease in Ca(++) levels in the endoplasmic reticulum (ER, [Ca(++)](ER)) observed five minutes following the initial challenge with PA. This decrease in [Ca(++) ](ER) was followed by a significant increase in the expression of ER stress signature genes CHOP, Xbp1 and GRP78. The early ER stress response induced by PA-LTx was followed by a strong mitochondrial membrane depolarization. Flow cytometry using 2\', 7\'-dichlorodihydrofluorescein diacetate (H(2)DCFDA) showed an increase in oxidative stress within three to six hours after PA treatment. Treatment of cultures undergoing PA-LTx with the calcium chelator BAPTA-AM and the anti-oxidant MCI-186 significantly reversed the lipotoxic effect by decreasing the generation of ROS and significantly increasing cell viability. We conclude that lipotoxicity in Schwann cells results in cellular dysfunction and cell death that involves a robust ER stress response, mitochondrial dysfunction and an augmented state of cellular oxidative stress (ASCOS).Copyright © 2010 Elsevier B.V. All rights reserved.

Keyword: hyperlipedemia

Insulin-independent regulation of hepatic triglyceride synthesis by fatty acids.

A central paradox in type 2 diabetes is the apparent selective nature of hepatic insulin resistance--wherein insulin fails to suppress hepatic glucose production yet continues to stimulate lipogenesis, resulting in hyperglycemia, , and hepatic steatosis. Although efforts to explain this have focused on finding a branch point in insulin signaling where hepatic glucose and lipid metabolism diverge, we hypothesized that hepatic triglyceride synthesis could be driven by substrate, independent of changes in hepatic insulin signaling. We tested this hypothesis in rats by infusing [U-(13)C] palmitate to measure rates of fatty esterification into hepatic triglyceride while varying plasma fatty and insulin concentrations independently. These experiments were performed in normal rats, high fat-fed insulin-resistant rats, and insulin receptor 2\'-O-methoxyethyl chimeric antisense oligonucleotide-treated rats. Rates of fatty esterification into hepatic triglyceride were found to be dependent on plasma fatty infusion rates, independent of changes in plasma insulin concentrations and independent of hepatocellular insulin signaling. Taken together, these results obviate a paradox of selective insulin resistance, because the major source of hepatic lipid synthesis, esterification of preformed fatty acids, is primarily dependent on substrate delivery and largely independent of hepatic insulin action.

Keyword: hyperlipedemia

Stereospecific distribution of fatty acids in human plasma triglycerides.

Because of a biological difference between both primary hydroxylgroups of glycerol there are three distinct hydroxylgroups esterified with fatty acids in triglycerides. The distribution of fatty acids in the three distinct positions of triglycerides can be investigated by a combination of partial hydrolysis of triglycerides by pancreatic lipase, phosphorylation of the diglycerides formed and the stereospecific action of phospholipase A on phosphatides. Plasma triglycerides from normal and hyperlipemic humans are analysed. Human plasma triglycerides are highly asymmetric. The 1-position contains more than twice as much as the 3-position. Statistically significant differences are found between normal and hyperlipemic humans for and linoleic at the 2-position, and for oleic and linoleic at the 3-position.

Keyword: hyperlipedemia

Saturated fat and lipemia: importance of study design and triglyceride structure.

Keyword: hyperlipedemia

Erythrocyte membrane fatty composition in hypercholesterolaemia.

The fatty composition of the erythrocyte membrane was determined in 22 hypercholesterolaemic patients managed by dietary restriction, and compared with that of 22 normocholesterolaemic controls, roughly matched for age and sex with the patient group. The patients exhibited higher relative proportions of (P < 0.01) and stearic (P < 0.005) acids and lower relative proportions of linoleic (P < 0.05) in the erythrocyte membrane, compared with controls, which could be due to presumed dietary differences between the groups. In the patients, the degree of hypercholesterolaemia was poorly correlated with erythrocyte linoleic . Measurement of erythrocyte linoleic might prove useful in the routine management of hypercholesterolaemia.

Keyword: hyperlipedemia

High density lipoprotein (HDL) reverses induced energy metabolism imbalance by switching CD36 and GLUT4 signaling pathways in cardiomyocyte.

In our previous study (PA) induced lipotoxicity and switches energy metabolism from CD36 to GLUT4 in H9c2 cells. Low level of high density lipoprotein (HDL) is an independent risk factor for cardiac hypertrophy. Therefore, we in the present study investigated whether HDL can reverse PA induced lipotoxicity in H9c2 cardiomyoblast cells. In this study, we treated H9c2 cells with PA to create a model in vitro and analyzed for CD36 and GLUT4 metabolic pathway proteins. CD36 metabolic pathway proteins (phospho-AMPK, SIRT1, PGC1α, PPARα, CPT1β, and CD36) were decreased by high PA (150 and 200\u2009μg/μl) concentration. Interestingly, expression of GLUT4 metabolic pathway proteins (p-PI3K and pAKT) were increased at low concentration (50\u2009μg/μl) and decreased at high PA concentration. Whereas, phospho-PKCζ, GLUT4 and PDH proteins expression was increased in a dose dependent manner. PA treated H9c2 cells were treated with HDL and analyzed for cell viability. Results showed that HDL treatment induced cell proliferation efficiency in PA treated cells. In addition, HDL reversed the metabolic effects of PA: CD36 translocation was increased and reduced GLUT4 translocation, but HDL treatment significantly increased CD36 metabolic pathway proteins and reduced GLUT4 pathway proteins. Rat neonatal cardiomyocytes showed similar results. In conclusion, HDL reversed palmatic -induced lipotoxicity and energy metabolism imbalance in H9c2 cardiomyoblast cells and in neonatal rat cardiomyocyte cells.© 2017 Wiley Periodicals, Inc.

Keyword: hyperlipedemia

Effect on plasma lipids of interesterifying a mix of edible oils.

The aim was to establish whether interesterification of oils, an alternative to the use of trans fatty acids in margarine manufacture, adversely affects plasma lipids. Twenty-seven hypercholesterolemic men participated in a double-blind, crossover trial of three margarines: 1) high-linoleic , moderate trans fatty acids; 2) high-palm oil blend (predominantly lauric, myristic, , oleic, and linoleic acids); and 3) an interesterified form of the high-palm oil margarine. Both high-palm oil margarines led to similar low-density-lipoprotein (LDL) cholesterol concentrations (4.43 +/- 0.94 and 4.54 +/- 0.88 mmol/L, respectively), which were significantly higher than the LDL concentrations after the high-linoleic margarine (4.02 +/- 0.85 mmol/L, P < 0.001). Interesterification transferred substantial proportions of into the sn-2 position of glycerides and unsaturated fatty acids into the sn-1,3 positions, and this was reflected in plasma chylomicrons. This study shows that interesterification of oils used to harden margarines does not raise plasma cholesterol more than does the margarine\'s constituent fatty acids.

Keyword: hyperlipedemia

[ induces apoptosis in human hepatoma cell line, HepG2 cells].

To investigate the effects of (PA) on human hepatocytes and its mechanism.We administered a mimic condition of 0.2-0.4 mmol/L PA to human hepatoma cell line, HepG2 cells. Cell viability was determined by Trypan blue staining. Cell cycle and early apoptosis were determined by propidium iodide and/or Annexin V staining, and the levels of Bcl-2 and Bax were analyzed by flow cytometry.An inhibition of cell growth was observed at a dose- and time-dependent manner in HepG2 cells after the treatment of PA. An apoptosis with appearance of sub-G1 fraction determined by cell cycle analysis significantly increased after the treatment of PA for 4 days. Bcl-2 level slightly decreased; in contrast, Bax level elevated markedly, which resulted in a significant decrease of Bcl-2/Bax ratio.PA may induce cell death on hepatocytes via mitochondria-mediated apoptosis by reducing the level of Bcl-2/Bax.

Keyword: hyperlipedemia

[The function of mitochondrion, carnitine, coenzyme-A, fat acids, glucose, the Randle cycle and insulin: a lecture].

In the function of feeding as biologic function of trophology, occurs the interchange of biologic reactions of exotrophy (postprandial hyperglycemia and ) and periods of food absence (biologic reaction of endotrophy). The action of insulin is realized in the biologic reaction of exotrophy only. The phylogenetic late insulin manages the function of phylogenetic early mitochondrion in the biologic reaction of exotrophy. In the biologic reaction of exotrophy, the cells activatedly absorb glucose under effect of insulin through glucose carriers-4 and actively absorb fat acids in the form of nonpolar triglycerides in olein lipoproteins of very low density by force of receptor endocytosis. These mechanisms formed on late stages of phylogenesis in the becoming of biologic function of locomotion--the function of movement. In the biologic reaction of endotrophy the cells absorb passively glucose under effect of glycaemia in intercellular medium through glucose carriers and passively (on gradient of concentration) absorb the fat acids in polar form under passive diffusion through plasmatic membrane. These reactions are worked out on the early stages of phylogenesis in the becoming of biologic function of homeostasis. The syndrome of resistance to insulin is formed only in biologic reaction of exotrophy but the disorders can persist and during realization of biologic reaction of endotrophy. In the biologic reaction of exotrophy insulin "decides": a) glucose can be deposited only in a limited way and can be consumed (oxidized in mitochondrion) in the first instance: b) fat acids can be stored and kept to be used in biologic function of locomotion. In the biologic reaction of exotrophy insulin "endeavors" as fast and full as possible use glucose and preserve in vivo as much as possible of fat acids as a substratum for further realization of biologic function of locomotion. Insulin minimizes in cytosol the content of a) ketone bodies - metabolites of C4 butyric fat and b) short chained C6-C10 fat acids and C16 for which in mitochondrion exists specific carrier - carnitin-palmitoilacyltransferase and "forces" mitochondrion to oxidize glucose. The main biologic role of insulin is to provide the biologic function of locomotion with substratum of energy.

Keyword: hyperlipedemia

[Lipid-reducing basic diet and serum fatty levels].

In 50 patients with primary hyperlipoproteinaemia during a 3 year treatment with lipid reducing metabolic basis diet in intervals of 4 months the composition of the cholesterol ester and triglyceride fatty acids of the serum was determined by means of gas chromatography. After 12--16 months in the hyperlipoproteinaemia type IIa (10 patients) in the cholesterol ester fraction, after 12 months in the hyperlipoproteinaemia type IIb (10 patients) in the cholesterol ester fraction and in the triglyceride fraction significant changes of the spectre of fatty acids developed. , stearic , palmitoleic , oleic and eikosatrienic decreased, and linolic , linolenic , arachidonic and eikosapentaenic increased. In the course of the further therapy the precental proportions of the fatty acids did no more change. Triglyceride fatty acids in hyperlipoproteinaemia of type IIa and cholesterol ester fatty acids in hyperlipoproteinaemia of type IIb were not influenced. These metabolic effects of the metabolic basis diet on the spectres of fatty acids of cholesterol ester and triglyceride fractions in primary hyperlipoproteinaemia of type IIa, IIb and IV must not be seen only in their increased content of polyene fatty acids which influences the hepatogenic processes of synthesis and oxydation of fatty acids, but are traced back also to the reduced proportions of fat, mono- and disaccharide. With metabolic basis diet decreasing lipids alone we, however, did not succeed in normalising fully the composition of fatty acids, and the constellations of fatty acids of test persons with healthy metabolism were reached only approximately.

Keyword: hyperlipedemia

Triacylglycerol structure and interesterification of and stearic -rich fats: an overview and implications for cardiovascular disease.

The position of fatty acids in the TAG molecule (sn-1, sn-2 and sn-3) determines the physical properties of the fat, which affects its absorption, metabolism and distribution into tissues, which may have implications for the risk of CHD. The TAG structure of fats can be manipulated by the process of interesterification, which is of increasing commercial importance, as it can be used to change the physical characteristics of a fat without the generation of trans-fatty acids. Interesterified fats rich in long-chain SFA are commercially important, but few studies have investigated their health effects. Evidence from animal and human infant studies suggests that TAG structure and interesterification affect digestibility, atherogenicity and fasting lipid levels, with fats containing and stearic in the sn-2 position being better digested and considered to be more atherogenic. However, chronic studies in human adults suggest that TAG structure has no effect on digestibility or fasting lipids. The postprandial effects of fats with differing TAG structure are better characterised but the evidence is inconclusive; it is probable that differences in the physical characteristics of fats resulting from interesterification and changes in TAG structure are key determinants of the level of postprandial lipaemia, rather than the position of fatty acids in the TAG. The present review gives an overview of TAG structure and interesterified and stearic -rich fats, their physical properties and their acute and chronic effects in human adults in relation to CHD.

Keyword: hyperlipedemia

[The role of modification of fatty composition of erythrocyte lipids in pathogenesis of arterial hypertension].

We used liquid chromatography for analysis of fatty acids (FA) in lipids of erythrocytes of patients with hypertensive disease (HD) with normo- (group 1) and (group 2). Abnormalities of FA composition of erythrocyte lipids were revealed in both groups. In group 1 we found deficit of polyenic acids of omega-6 family, accumulation of Mead - prostanoid precursor with pronounced vasoconstrictor and pro inflammatory properties. In group 2 we noted more profound rearrangement of lipid matrix of erythrocyte membrane manifested as deficiency of omega-3 polyenic acids, accumulation of palmitinic and arachidonic acids. Preponderance of saturated FA in erythrocytes and deficiency of polyenic acids might evidence for pathology of their ligand-receptor transport into the cell. Blockade of active FA transport initiates formation of HD, promotes accumulation of atherogenic fractions of lipoproteins in blood. These results evidence for important pathogenetic role of FA in development of hypertension.

Keyword: hyperlipedemia

Influence of pantethine on platelet volume, microviscosity, lipid composition and functions in diabetes mellitus with .

The in vivo effects of pantethine were investigated on serum lipids and platelet lipid and platelet functions in 31 diabetic patients with . Pantethine decreased cholesterol from 236 +/- 62 mg/dl (M +/- SD) to 217 +/- 51 mg/dl (p less than 0.01) and increased high density lipoprotein cholesterol from 40 +/- 11mg/dl to 43 +/- 15 mg/dl. The diabetic platelets were larger when accompanied by higher microviscosity that healthy platelets. The characteristics of lipid composition in diabetic platelets were high levels of free cholesterol, phospholipid, triglyceride, cholesterol ester, palmitoleic , linoleic and palmitoleic / and low levels of the molar ratio of free cholesterol/phospholipids, phosphatidylethanolamine, oleic , arachidonic and oleic /stearic . Pantethine normalized these values of fatty acids to the control levels, and concomitantly reduced significantly the hyperaggregation of platelets induced by 10(6) M ADP and the hyper-ADP release reaction from platelets when exposed to 2 microgram of collagen, and made the volume smaller and the microviscosity lower after oral administration. From these data, it was concluded that pantethine normalized the abnormalities of serum lipids as well as platelet lipid compositions and subsequently reduced the hyper-aggregation and hyper-release reactions through the changes of volume and microviscosity of the platelets in diabetes mellitus with .

Keyword: hyperlipedemia

Procyanidin B2 ameliorates free fatty acids-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state.

Procyanidin B2, a naturally occurring phenolic compound, has been reported to exert multiple beneficial functions. However, the effect of procyanidin B2 on free fatty acids (FFAs)-induced hepatic steatosis remains obscure. The present study is therefore aimed to elucidate the protective effect of procyanidin B2 against hepatic steatosis and its underlying mechanism. Herein, we reported that procyanidin B2 attenuated FFAs-induced lipid accumulation and its associated oxidative stress by scavenging excessive ROS and superoxide anion radicals, blocking loss of mitochondrial membrane potential, restoring glutathione content, and increasing activity of antioxidant enzymes (GPx, SOD and CAT) in hepatocytes. Procyanidin B2 mechanistically promoted lipid degradation via modulation of transcription factor EB (TFEB), a master regulator of lysosomal pathway. Molecular docking analysis indicated a possible ligand-binding position of procyanidin B2 with TFEB. In addition, administration of procyanidin B2 resulted in a significant reduction of hepatic fat accumulation in high-fat diet (HFD)-induced obese mice, and also ameliorated HFD-induced metabolic abnormalities, including and hyperglycemia. It was confirmed that procyanidin B2 prevented HFD-induced hepatic fat accumulation through down-regulating lipogenesis-related gene expressions (PPARγ, C/EBPα and SREBP-1c), inhibiting pro-inflammatory cytokines production (IL-6 and TNF-α) and increasing antioxidant enzymes activity (GPx, SOD and CAT). Moreover, hepatic fatty acids analysis indicated that procyanidin B2 caused a significant increase in the levels of , oleic and linoleic . Intriguingly, procyanidin B2 restored the decreased nuclear TFEB expression in HFD-induced liver steatosis and up-regulated its target genes involved in lysosomal pathway (Lamp1, Mcoln, Uvrag), which suggested a previously unrecognized mechanism of procyanidin B2 on ameliorating HFD-induced hepatic steatosis. Taken together, our results demonstrated that procyanidin B2 attenuated FFAs-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state, which had important implications that modulation of TFEB might be a potential therapeutic strategy for hepatic steatosis and procyanidin B2 could represent a promising novel agent in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD).Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering via regulating the AMPK/SREBP1/PPARα signaling cascade of lipid metabolism.

The present study for the first time aims to examine the hypothesis that circulating gamma-glutamyl carboxylated growth arrest specific protein 6 (Gla-Gas6) deficiency may be associated with and vitamin K (VK) supplementation may ameliorate the impaired lipid homeostasis via activating Gas6 protein. Subjects with (n=22) and age-matched healthy controls (n=19) were included in this study. Results showed that plasma levels of Gla-Gas6 protein and VK were significantly lower in hyperlipidemic subjects compared to control. Moreover, Gla-Gas6 levels were significantly and positively correlated with VK (P=.034, r=0.452) and negatively with triglyceride (P=.022, r=-0.485) and total cholesterol (P=.043, r=-0.435) in hyperlipidemic subjects, which suggest that VK supplementation may have a positive effect in activating Gas6 protein and thereby reducing the aberrant plasma lipid levels. Further studies with high-fat diet (HFD)-fed animal model of demonstrated that VK supplementation (5 μg/kg body weight, 8 weeks) reduced the plasma lipid levels, stimulated both the plasma levels and the hepatic protein expression of Gla-Gas6 protein, and regulated the AMPK/SREBP1/PPARα signaling pathways of hepatic lipid metabolism in HFD-fed mice. Moreover, by using (PA, 0.75 mM)-treated both control and GGCX knockdown hepatocytes, this study dissected the direct role of Gla-Gas6 in mediating the positive effect of VK on preventing the PA-induced impaired hepatic lipid metabolism via regulating AMPK/SREBP1/PPARα pathways. Combining all, the present study demonstrated the beneficial effect of VK supplementation in preventing the impaired lipid homeostasis via activating VK-dependent Gas6 protein.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

The triglyceride-lowering effect of supplementation with dual probiotic strains, Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032: Reduction of fasting plasma lysophosphatidylcholines in nondiabetic and hypertriglyceridemic subjects.

This study evaluated the triglyceride (TG)-lowering effects of consuming dual probiotic strains of Lactobacillus curvatus (L. curvatus) HY7601 and Lactobacillus plantarum (L. plantarum) KY1032 on the fasting plasma metabolome.A randomized, double-blind, placebo-controlled study was conducted on 92 participants with hypertriglyceridemia but without diabetes. Over a 12-week testing period, the probiotic group consumed 2 g of powder containing 5 × 10(9) colony-forming units (cfu) of L. curvatus HY7601 and 5 × 10(9) cfu of L. plantarum KY1032 each day, whereas the placebo group consumed the same product without probiotics. Fasting plasma metabolomes were profiled using UPLC-LTQ-Orbitrap MS. After 12 weeks of treatment, the probiotic group displayed a 20% reduction (p = 0.001) in serum TGs and 25% increases (p=0.001) in apolipoprotein A-V (apoA-V). At the 12-week follow-up assessment, the following 11 plasma metabolites were significantly reduced in the probiotic group than the placebo group: palmitoleamide, amide, oleamide, and lysophosphatidyl choline (lysoPC) containing C14:0, C16:1, C16:0, C17:0, C18:3, C18:2, C18:1, and C20:3. In the probiotic group, changes (▵) in TG were negatively correlated with ▵ apoA-V, which was positively correlated with ▵ FFA. In addition, ▵ FFA was strongly and positively correlated with ▵ lysoPCs in the probiotic group but not the placebo group.The triglyceride-lowering effects of probiotic supplementation, partly through elevated apoA-V, in borderline to moderate hypertriglyceridemic subjects showed reductions in plasma metabolites, fatty primary amides and lysoPCs (; http://www.clinicaltrials.gov). Clinical trials: ; http://www.clinicaltrials.gov.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: hyperlipedemia

Investigation of 3,5-isoxazolidinediones as hypolipidemic agents in rodents.

A series of 2-benzoyl-4,4-dialkyl-3,5-isoxazolidinediones proved to have potent hypolipidemic activity, lowering both serum cholesterol and triglyceride levels at 10 or 20 mg/kg/day, IP and orally in rodents. 2-(3,4,5-Trimethoxybenzoyl)-4,4-diethyl-3,5-isoxazolidinedione+ ++ (4) afforded the best hypolipidemic activity lowering normolipidemic CF1 mouse serum cholesterol levels 49% and serum triglyceride levels 34% at 20 mg/kg/day, IP. Compound 4 was selected as a typical derivative of the chemical class for further detailed studies. Serum cholesterol levels in normolipidemic Sprague Dawley male rats were reduced 45% after 8 weeks at 10 and 20 mg/kg/day of compound, orally. Serum triglyceride levels were reduced 38-49% at 10 and 20 mg/kg/day, orally. In vitro liver enzyme activities studies in normolipidemic CF1 mice showed the compound inhibited mitochondrial citrate exchange, acetyl CoA synthetase, HMG CoA reductase, acyl CoA cholesterol acyl transferase, acetyl CoA carboxylase, sn-glycerol-3-phosphate acyl transferase, phosphatidylate phosphohydrolase and heparin-induced lipoprotein lipase activities with increases in the activities of cholesterol ester hydrolase and ATP-dependent citrate lyase. Similar enzyme activities were inhibited in vivo except HMG CoA reductase activity was not inhibited in rat liver or small intestinal mucosa after 8 weeks drug administration. Cholesterol levels were reduced in tissues after 8 weeks administration of compound 4 in normolipidemic rats. Bile cholesterol and triglyceride levels were elevated after two weeks administration to rats at 20 mg/kg/day. Serum lipoprotein levels in normolipidemic and hyperlipidemic rats showed the cholesterol levels in VLDL and LDL fractions after 4, 6 and 8 weeks at 10 and 20 mg/kg/day were reduced whereas HDL-cholesterol levels were significantly elevated. Studies demonstrated that 3H-cholesterol and 14C- incorporation into lipids of the lipoprotein fraction was reduced by the drug but 32P-incorporation was generally elevated. The agent demonstrated no observable toxicity in rats after 8 weeks administration, orally. The acute toxicity study in normolipidemic mice at 20, 40 and 100 mg/kg/day, IP, demonstrated no observable harmful effects of the drug.

Keyword: hyperlipedemia

Supplementation with vitamin E in hyperlipidemic patients treated with diet and clofibrate. Effects on serum lipoprotein concentrations, plasma fatty composition and adipose tissue lipoprotein lipase activity.

Twelve hyperlipidemic patients on long term treatment with a lipid lowering diet enriched in polyunsaturated fatty acids and with clofibrate were supplemented with vitamin E (400 mg/day). The effect on serum lipoprotein concentration, plasma lipid fatty composition, and adipose tissue lipoprotein lipase activity was studied. No additional lipid-lowering effect was registered during a treatment period of 4 months. A slight increase in total serum cholesterol concentration and in high density lipoprotein concentration was probably attributable to seasonal variations in serum lipoprotein concentrations. No major changes of fatty composition in plasma cholesteryl esters or triglycerides were recorded. However, an increased relative amount of arachidonic and a reduced amount of in the plasma phospholipids after 2 months was possibly caused by the vitamin E therapy.

Keyword: hyperlipedemia

-induced hepassocin in the liver contributes to insulin resistance in skeletal muscle.

Hepassocin (HPS) has recently been identified as a novel hepatokine that causes hepatic steatosis. However, the role of HPS in the development of insulin resistance in skeletal muscle under obesity remains unclear. The effect of on hepatic HPS expression was evaluated in primary hepatocytes and liver of mice. HPS-mediated signal pathways were explored using small interfering (si) RNAs of specific genes or inhibitors. We found that treatment of primary hepatocytes with palmitate could induce HPS expression through C/EBPβ-mediated transcriptional activation. Furthermore, increased HPS expression was observed in the liver of high fat diet (HFD)-fed or tunicamycin-treated mice. Pretreatment with 4-phenylbutyrate (4-BPA) (an endoplasmic reticulum (ER) stress inhibitor) and suppression of p38 by siRNA abrogated the effect of palmitate on HPS expression in primary hepatocytes. Treatment of differentiated C2C12\xa0cells with recombinant HPS caused c-Jun N-terminal kinase (JNK) phosphorylation and impairment of insulin sensitivity in a dose-dependent manner. siRNA-mediated suppression of JNK reduced the effect of HPS on insulin signaling. Furthermore, the suppression of epidermal growth factor receptor (EGFR) by siRNA mitigated both HPS-induced JNK phosphorylation and insulin resistance. In addition, HPS did not affect inflammation and ER stress in differentiated C2C12\xa0cells. In conclusion, we elucidated that ER stress induced by palmitate could increase the expression of HPS in hepatocytes and further contribute to the development of insulin resistance in skeletal muscle via EGFR/JNK-mediated pathway. Taken together, we suggest that HPS could be a therapeutic target for obesity-linked insulin resistance.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: hyperlipedemia

An interesterified palm olein test meal decreases early-phase postprandial lipemia compared to palm olein: a randomized controlled trial.

Palm oil that has been interesterified to produce a higher proportion of (16:0) in the sn-2 position reduces postprandial lipemia in young, normolipidemic men and women, but effects in older subjects with higher fasting triacylglycerol (TAG) concentrations are unknown. We tested the hypothesis that high-fat meals rich in interesterified palm olein (IPO) decrease lipemia and alter plasma lipoprotein fraction composition compared to native palm olein (NPO) in men aged 40-70 years with fasting TAG concentrations ≥1.2 mmol/L. Postprandial changes in plasma lipids following meals containing 75 g fat (NPO and IPO) were compared using a randomized, double-blind crossover design (n = 11). Although there were no significant differences in plasma TAG concentrations between meals over the total 6-h postprandial measurement period, IPO resulted in a decreased plasma TAG response during the first 4 h of the postprandial period (iAUC 1.65 mmol/L h, 95% CI 1.01-2.29) compared to NPO (iAUC 2.33 mmol/L h, 95% CI 1.58-3.07); meal effect P = 0.024. Chylomicron fraction TAG concentrations at 4-6 h were slightly reduced following IPO compared to NPO [NPO-IPO mean difference 0.29 mmol/L (95% CI -0.01-0.59), P = 0.055]. There were no differences in IDL fraction TAG, cholesterol or apolipoprotein B48 concentrations following IPO compared with NPO. In conclusion, consuming a meal containing palm olein with a higher proportion of 16:0 in the sn-2 position decreases postprandial lipemia compared to native palm olein during the early phase of the postprandial period in men with higher than optimal fasting triacylglycerol concentrations.

Keyword: hyperlipedemia

Fatty acids and glucose increase neutral endopeptidase activity in human microvascular endothelial cells.

Neutral endopeptidase (NEP), a membrane-bound metallopeptidase enzyme that degrades neuropeptides, bradykinin, atrial natriuretic factor, enkephalins, and endothelin may regulate response to injury. We have previously demonstrated increased NEP localization and enzyme activity in diabetic wounds and skin compared with normal controls. We hypothesized that and hyperglycemia associated with type 2 diabetes mellitus may induce excessive NEP activity and thereby diminish normal response to injury. Human microvascular endothelial cells were treated with five different fatty acids (40 microM) with varying degrees of saturation, including oleic , linoleic , , stearic , and linolenic and/or glucose (40 mM) for 48 h. The effect of the antioxidative agents vitamin E and C on NEP enzyme activation was determined by treating the cultured cells with alpha-tocopherol succinate and/or L-ascorbic . Cell membrane preparations were assayed for NEP activity by incubation with glutaryl-Ala-Ala-Phe-4-methoxy-beta naphthylamide to generate a fluorescent degradation product methoxy 2 naphthylamine. High glucose or fatty concentration upregulated NEP activity. The highest NEP activity was observed with combined elevated glucose, linoleic , and oleic (P < 0.05). Antioxidant vitamin E and C treatment significantly reduced NEP enzyme activity after fatty exposure (P < 0.05). Thus, hyperglycemia and associated with type 2 diabetes mellitus may increase endothelial cell NEP activity and thereby decrease early pro-inflammatory responses. The modulator effect of vitamin E and C on NEP membrane enzyme activity after exposure to fatty stimulation suggests that lipid oxidation may activate NEP.

Keyword: hyperlipedemia

Liver X receptor agonist T0901317 enhanced peroxisome proliferator-activated receptor-delta expression and fatty oxidation in rat skeletal muscle.

Liver X receptors (LXR) have been characterized as key transcriptional regulators of hepatic lipid and carbohydrate metabolism. LXR are expressed also in skeletal muscle, however, their role in this tissue is poorly investigated and the vast majority of available data comes from studies on cultured myotubes. Therefore, we aimed to examine effects of in vivo LXR activation on muscle lipid metabolism. The experiments were performed on male Wistar rats fed on a standard rodent chow. The animals were divided into two groups (n=10) receiving either LXR activator (T0901317, 10 mg/kg/day) or vehicle for one week. Samples of the soleus as well as red and white sections of the gastrocnemius muscle were excised. T0901317 increased muscle expression of peroxisome proliferator-activated receptor-δ and its target genes involved in fatty uptake and oxidation. In addition, LXR agonist enhanced palmitate oxidation (by 55%) in isolated soleus muscle. However, palmitate incorporation into triacylglycerol was decreased (by 38%), which was associated with reduced diacylglycerol acyltransferase expression (by 66%). Despite markedly increased plasma lipid concentration upon T0901317 treatment, muscle triacylglycerol level was elevated only in the red section of the gastrocnemius muscle. We conclude that T0901317 enhances muscle fatty oxidation, which prevents overt accumulation of intramuscular lipids that could be expected considering T0901317-induced .

Keyword: hyperlipedemia

Role of protein kinase C isoenzymes in fatty stimulation of insulin secretion.

Although is frequently associated with hyperinsulinemia. the stimulation of insulin secretion by fatty acids in the in vitro studies has remained a matter of constant debate, partly because of the uncertainty about a clearly defined mechanism to explain such a direct effect. In this study, we used a pharmacologic approach to test the hypothesis that protein kinase C (PKC) signal-transduction pathway is involved in fatty -stimulated insulin secretion. Isolated rat islets were perifused with either palmitate (C(16:0)) or linoleate (C(18:2)) in the absence or presence of selective inhibitors of PKC isoenzymes. Our results suggest a role for Ca2+-independent PKC isoenzymes in the signal transduction of fatty -stimulated insulin secretion. The data imply that either the nonconventional and/or atypical isoforms of PKC are involved in the stimulation of insulin release induced by fatty acids.

Keyword: hyperlipedemia

Lecithin has no effect on serum lipoprotein, plasma fibrinogen and macro molecular protein complex levels in hyperlipidaemic men in a double-blind controlled study.

To examine the effects of lecithin on serum lipoprotein, plasma fibrinogen and macro molecular protein complex (MPC) levels.Twenty free living hyperlipidaemic men participated in this double-blind study which controlled for possible indirect effects. The subjects were randomly assigned to one of three treatments: frozen yoghurt or frozen yoghurt with 20 g soya bean lecithin or frozen yoghurt with 17 g sunflower oil. Sunflower oil was used to control for the increased energy and linoleic intake from lecithin. Yoghurt served as the \'vehicle\' for the lecithin and sunflower oil and yoghurt alone was given to one group to control for possible effects due to the yoghurt \'vehicle\', as well as other environmental influences. Variables were measured with standard methods twice at baseline and after 2 and 4 weeks of treatment.Plasma linoleic levels increased significantly with lecithin and sunflower oil treatments indicating that compliance to the treatments were obtained. Lecithin treatment did not have significant effects on serum total cholesterol, triglyceride, high density lipoprotein cholesterol, low density lipoprotein cholesterol, apolipoprotein A, apolipoprotein B or lipoprotein (a) levels. Plasma fibrinogen and MPC levels were also not affected by lecithin therapy. Sunflower oil treatment resulted in significant increased body weight, serum TC and decreased MPC levels.Lecithin treatment had no independent effects on serum lipoprotein, plasma fibrinogen or MPC levels in hyperlipidaemic men.

Keyword: hyperlipedemia

Cannabinoid receptor 1 mediates -induced apoptosis via endoplasmic reticulum stress in human renal proximal tubular cells.

The endocannabinoid system (ECS) is activated at the onset of obesity and diverse metabolic diseases. Endocannabinoids mediate their physiological and behavioral effects by activating specific cannabinoid receptors, mainly cannabinoid receptor 1 (CB(1)R). Diabetic nephropathy (DN) is induced by , and renal proximal tubule cells are an important site for the onset of DN. However, the pathophysiology of CB(1)R, especially in the of DN, has not been elucidated. Therefore, we examined the effect of (PA) on CB(1)R expression and its related signal pathways in human renal proximal tubular cells (HK-2 cells). PA significantly increased CB(1)R mRNA and protein levels and induced CB(1)R internalization. PA-induced activation of CB(1)R is prevented by the treatment of AACOCF(3) (a cPLA(2) inhibitor), indomethacin and NS398 (a COX 2 inhibitors). Indeed, PA increased cPLA(2), and COX-2 but not COX-1. We also investigated whether the PA-induced activation of CB(1)R is linked to apoptosis. As a result, AM251 (a CB(1)R antagonist) attenuated PA-mediated apoptosis in a concentration-dependent manner. Furthermore, PA decreased GRP78 expression and induced increases in the endoplasmic reticulum (ER) stress signaling pathways p-PERK, p-eIF2α, p-ATF4, and CHOP, which were blocked by AM251 treatment. Moreover, PA increased the Bax/Bcl-2 ratio, cleaved PARP, and caspase-3 levels. The PA-induced apoptotic effects were decreased with CB(1)R-specific antagonist (AM251) treatment and CB1 si-RNA transfection. In conclusion, PA induced apoptosis through ER stress via CB(1)R expression in human proximal tubule cells. Our results provide evidence that CB(1)R blockade may be a potential anti-diabetic therapy for the treatment of DN.© 2010 Wiley-Liss, Inc.

Keyword: hyperlipedemia

Nonpharmacological approaches for reducing serum low-density lipoprotein cholesterol.

To reinforce the key role of diet and lifestyle modification as the first-line treatment for the reduction of raised serum low-density lipoprotein cholesterol (LDL-C) and prevention of cardiovascular disease. Also, to counter recent claims that the current dietary guidelines for the treatment of cardiovascular disease have misplaced emphasis on the importance of removing dietary saturated fat instead of sugar.This review provides new insight into the effects of diet and lifestyle factors with established efficacy in lowering serum LDL-C. This includes energy-restricted weight loss and new findings on the effects of alternative day fasting; novel metabolic and molecular effects of replacing with oleic ; evidence for a dose-response relationship between the intake of dietary stanols and LDL-C; and identification of a unique metabolic pathway for the excretion of cholesterol.The review reports new evidence for the efficacy of alternate day fasting, reassurance that the current dietary guidelines are not misguided by recommending removal of saturated fat, that a high intake of dietary stanols can achieve a reduction in LDL-C of up to 18%, and describes a pathway of cholesterol excretion that may help to explain variation in the response of serum LDL-C to dietary fat and cholesterol.

Keyword: hyperlipedemia

[The alternative view on diagnostic of hyperlipoproteinemia, cholesterol lipoproteins of low density and effect of statins: a lecture].

The effect of statins occur in several stages: 1) inhibition in hepatocytes of synthesis of functionally specific pool of spirit cholesterol, polar mono-layer of lipoproteins of very low density; 2) activation of hydrolysis of triglycerides in lipoproteins of very low density, formation of apoE/B-100-ligand and absorption of lipoproteins of very low density by insulin-depended cells; 3) decreasing of content of and spirit cholesterol-lipoproteins of very low density in blood plasma; 4) activation of hydrolysis of triglycerides in lipoproteins of low density, formation of apoB-100-ligand and absorption of lipoproteins of low density by insulin-independent cells; 5) decreasing of level of and increasing of content of lipoproteins of high density. During first weeks of effect of statins occurs decreasing of concentration of triglycerides and unesterified spirit cholesterol-lipoproteins of very low density in blood plasma. Then, slower and more durational decreasing of level of spirit cholesterol-lipoproteins of low density occurs. The value of spirit cholesterol-lipoproteins of low density is primarily determined by content of saturated fatty in food, its endogenous synthesis from glucose and concentration of triglycerides and lipoproteins of very low density of the same name in blood plasma. The effect of preparations is biologically valid and corresponds to alternative hypolipidemic preparations. All these preparations have an effect following a common algorithm: they activate, using different mechanisms, receptor absorption of lipoproteins of very low density or lipoproteins of low density by cells. The level of spirit cholesterol-lipoproteins of low density in full measure depends on content of triglycerides in blood. The concentration of spirit cholesterol in blood plasma has a reliable diagnostic significance only under physiological content of triglycerides. The main criterion of diagnostic and control of hypolipidemic therapy biologically is content of triglycerides. The comprehension of differences in effect of hypolipidemic preparations within framework of common algorithm permits rationally combine them under treatment of both primary inheritable phenotypes of glucolipoproteins and secondary symptomatic types of glucolipoproteins under obligatory observation of strict dietary treatment.

Keyword: hyperlipedemia

Lipotoxicity in renal proximal tubular cells: relationship between endoplasmic reticulum stress and oxidative stress pathways.

in the general population has been linked to the development of chronic kidney disease with both oxidative and endoplasmic reticulum stress implicated. Physiological levels (50-300 micromol/L) of saturated fatty acids such as (PA) cause cytotoxicity in vitro. We investigated cell type- and stimulus-specific signaling pathways induced by PA in renal proximal tubular cells and whether oxidative stress leads to ER stress or vice versa and which pathways predominate in signaling for PA-induced apoptosis and necrosis. NRK-52E cells were incubated with PA or hydrogen peroxide (H(2)O(2)) combined with SP600125 which blocks c-Jun N-terminal kinase (JNK) activation; salubrinal, which maintains eukaryotic initiation factor 2 alpha in its phosphorylated state and the antioxidant EUK-134 - a superoxide dismutase mimetic with catalase activity. We found that (i) PA causes both oxidative and ER stress leading to apoptosis which is mediated by phosphorylated JNK; (ii) oxidant-induced apoptosis generated by H(2)O(2) involves ER stress signaling and CHOP expression; (iii) the ER stress mediated by PA is largely independent of oxidative stress; (iv) in contrast, the apoptosis produced by PA is mediated partly via oxidative stress. PA-mediated cell signaling in renal NRK-52E cells therefore differs from that identified in neuronal, hepatic and pancreatic beta cells.(c) 2010 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Dietary fatty acids, cholesterol, and the lipoprotein profile.

Keyword: hyperlipedemia

Blockade of myeloid differentiation protein 2 prevents obesity-induced inflammation and nephropathy.

Obesity is a major and independent risk factor of kidney diseases. The pathogenic mechanisms of obesity-associated renal injury are recognized to at least involve a lipid-rich and pro-inflammatory state of the renal tissues, but specific mechanisms establishing causal relation remain unknown. Saturated fatty acids are elevated in obesity, and known to induce chronic inflammation in kidneys. Myeloid differentiation protein 2 (MD2) is an important protein in lipopolysaccharide-induced innate immunity response and inflammation. We suggested that obesity-associated renal injury is regulated by MD2 thereby driving an inflammatory renal injury. The used three mouse models for in vivo study: MD2 knockout mice (KO) maintained on high fat diet (HFD), wild-type mice on HFD plus L6H21, a specific MD2 inhibitor and KO mice given (PA) by IV injection. The in vitro studies were carried out in cultured renal tubular epithelial cells, mouse mesangial cells and primary macrophages, respectively. The HFD mice presented with increased , serum creatinine and proteinuria. Renal tissue from HFD mice had increased fibrosis, inflammatory cytokines, macrophage infiltration, and activation of NF-κB and MAPKs. This HFD-induced renal injury profile was not observed in KO mice or L6H21-treated mice. Mice given PA mimmicked the HFD-induced renal injury profiles, which were prevented by MD2 knockout. The in vitro data further confirmed MD2 mediates PA-induced inflammation. MD2 is causally related with obesity-associated renal inflammatory injury. We believe that MD2 is an attractive target for future therapeutic strategies in obesity-associated kidney diseases.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: hyperlipedemia

Simultaneous measurement of free and esterified fatty acids by gas chromatography from normal and type IV hyperlipoproteinaemic sera.

A transmethylated reaction of esterified fatty acids with sodium- methoxide in a mixture of serum, petroleum ether and methanol is presented. In the conditions used the free fatty acids in the sample were not esterified. 1-2 microliters of the organic phase was injected into an OV-351 fused silica capillary column of gas liquid chromatography (GLC) fitted with flameionizing detector (FID) temperature program, and calculating integrator. By this method both free and esterified fatty acids were measured in a single run. The development of the present method for the better quantitation of free fatty acids is in progress as using gas chromatography/mass spectrometry for identification of the peaks. In thirteen healthy subjects, the serum free fatty content was 538 +/- 176 mumol/l and free glycerol 107 +/- 39 mumol/l, while in seven type IV hyperlipoproteinaemic sera the corresponding values were 1049 +/- 529 and 86 +/- 38 mumol/l. The most prominent differences between healthy and type IV hyperlipoproteinaemic subjects for esterified fatty acids were found in , oleic and arachidonic acids. The correlations between free and bound fatty acids has been discussed.

Keyword: hyperlipedemia

Impaired triacylglycerol catabolism in hypertriglyceridemia of the diabetic, cholesterol-fed rabbit: a possible mechanism for protection from atherosclerosis.

The etiology of the hypertriglyceridemia in alloxan-diabetic rabbits was studied by two independent methods. Production and removal rates of VLDL triacylglycerol were measured in diabetic rabbits by injection of [3H]palmitate-labelled VLDL. Similarly, triacylglycerol total removal rates were determined in non-diabetic rabbits which were infused with Intralipid to mimic the plasma triacylglycerol concentrations of diabetic rabbits. Compared to nondiabetic rabbits, triacylglycerol removal rats were decreased in diabetic rabbits, particularly at higher levels of plasma triacylglycerol. During cholesterol and triacylglycerol supplementation of the diet, post-heparin plasma lipoprotein lipase activity of diabetic rabbits with severe hypertriglyceridemia averaged 36% of that of nondiabetics, suggesting an impaired triacylglycerol removal capacity. Furthermore, plasma triacylglycerol was inversely related to post-heparin plasma lipoprotein lipase activity among diabetic rabbits. VLDL triacylglycerol production increased with increasing plasma triacylglycerol concentration among diabetic cholesterol-fed rabbits with moderately severe hypertriglyceridemia, but reached an apparent plateau among rabbits with plasma triacylglycerol concentrations from approx. 2000-9000 mg/dl. Thus, severe hypertriglyceridemia in this model of insulin deficiency can be attributed only partially to VLDL hypersecretion, whereas a removal defect, resulting in saturation of the triacylglycerol removal mechanism, appears to be largely responsible. The impaired removal of plasma triacylglycerol is also related to the presence of cholesterol predominantly in lipoproteins of increased size. The data support the hypothesis that protection against atherosclerosis in cholesterol-fed diabetic rabbits results from exclusion of very large cholesterol-containing lipoproteins from the arterial wall.

Keyword: hyperlipedemia

Inhibition of cholinergic potentiation of insulin secretion from pancreatic islets by chronic elevation of glucose and fatty acids: Protection by casein kinase 2 inhibitor.

Chronic and hyperglycemia are characteristic features of type 2 diabetes (T2DM) that are thought to cause or contribute to β-cell dysfunction by "glucolipotoxicity." Previously we have shown that acute treatment of pancreatic islets with fatty acids (FA) decreases acetylcholine-potentiated insulin secretion. This acetylcholine response is mediated by M3 muscarinic receptors, which play a key role in regulating β-cell function. Here we examine whether chronic FA exposure also inhibits acetylcholine-potentiated insulin secretion using mouse and human islets.Islets were cultured for 3 or 4 days at different glucose concentration with 0.5\xa0mM (PA) or a 2:1 mixture of PA and oleic (OA) at 1% albumin (PA/BSA molar ratio 3.3). Afterwards, the response to glucose and acetylcholine were studied in perifusion experiments.FA-induced impairment of insulin secretion and Ca signaling depended strongly on the glucose concentrations of the culture medium. PA and OA in combination reduced acetylcholine potentiation of insulin secretion more than PA alone, both in mouse and human islets, with no evidence of a protective role of OA. In contrast, lipotoxicity was not observed with islets cultured for 3 days in medium containing less than 1\xa0mM glucose and a mixture of glutamine and leucine (7\xa0mM each). High glucose and FAs reduced endoplasmic reticulum (ER) Ca storage capacity; however, preserving ER Ca by blocking the IP3 receptor with xestospongin C did not protect islets from glucolipotoxic effects on insulin secretion. In contrast, an inhibitor of casein kinase 2 (CK2) protected the glucose dependent acetylcholine potentiation of insulin secretion in mouse and human islets against glucolipotoxicity.These results show that chronic FA treatment decreases acetylcholine potentiation of insulin secretion and that this effect is strictly glucose dependent and might involve CK2 phosphorylation of β-cell M3 muscarinic receptors.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: hyperlipedemia

Antiretroviral therapy with or without protease inhibitors impairs postprandial TAG hydrolysis in HIV-infected men.

Mechanisms underlying the lipodystrophy syndrome associated with antiretroviral therapy (ART) for HIV infection are not completely understood. We investigated the effect of ART on blood lipid concentrations in the fasting state and after consumption of a meal containing [1-13C] in HIV-positive men receiving nucleoside reverse transcriptase inhibitors (NRTI, n 7), NRTI combined with protease inhibitors (PI; NRTIPI, n 6), in HIV-positive but therapy-naïve men (noART, n 5) and in HIV-seronegative men (controls, n 6). HIV-positive subjects had higher fasting TAG concentrations and resting energy expenditure than controls. Subjects receiving NRTIPI therapy had higher fasting NEFA concentrations than the other groups. There were no significant differences in postprandial lipid metabolism between noART subjects and controls. NRTI therapy impaired hydrolysis of meal-derived TAG, most evidently when combined with PI (the NRTIPI group). Accumulation of 13C-label in the NEFA fraction was not different between groups. In the NRTIPI group, fasting and postprandial NEFA concentrations were significantly higher than other groups. Postprandial glucose and insulin responses in HIV-positive subjects did not differ from controls. These findings suggest that ART dyslipidaemia is associated with impaired postprandial TAG clearance, which is exacerbated by NRTIPI therapy. If dyslipidaemia is to be minimised in ART, the specific adverse effects of particular combinations during the fed state should be considered.

Keyword: hyperlipedemia

Gestational in the rat is characterized by accumulation of n - 6 and n - 3 fatty acids, especially docosahexaenoic .

We have evaluated the relative and quantitative changes in long-chain fatty acids in maternal liver, serum, carcass and conceptus (fetuses plus placentae) during pregnancy in the rat, to ascertain whether previous concern over lower proportions of n - 6 and n - 3 fatty acids in maternal serum could be indicative of suboptimal n - 6 or n - 3 fatty status. Gestational was characterized by proportional decreases in linoleic, stearic and arachidonic acids but increases in and docosahexaenoic acids. However, the quantitative amount (microgram/ml) of linoleic, arachidonic and docosahexaenoic acids in serum lipids actually increased 2-5-fold from mid-pregnancy to term. Compared to non-pregnant rats, gestational was also associated with a lower proportion but similar quantity of linoleic in maternal carcass and adipose stores. We conclude that gestational in the rat is characterized by a relative but not quantitative decrease in whole-body stores of n - 6 fatty acids and a marked proportional and quantitative increase in docosahexaenoic in maternal organs and in the conceptus.

Keyword: hyperlipedemia

Fatty and lipid composition of the monkey retina in diet-induced hypercholesterolemia.

We determined the fatty composition of the lipids of cynomolgus monkey retina in animals fed commercial chow or a saturated fat, cholesterol-enriched atherogenic diet for 100 days. Doxosahexaenoic (22 : 6) accounted for 25.8% of the ethanolamine phosphoglyceride fatty acids, 17.6% of the serine plus inositol phosphoglyceride fatty acids, 8.4% of the choline phosphoglyceride fatty acids and 5.8% of the neutral lipid fatty acids in the retinas of the chow-fed animals. Therefore, monkey retinas, like those of other mammalian species, ordinarily contain large amounts of 22 : 6. Retinas from the monkeys fed the atherogenic diet contained less 22 : 6 as well as other polyunsaturates in each of the phospholipid classes. The decrease in polyunsaturates was compensated for by increases in , stearic, and oleic acids. There was no difference in the amount of phospholipid, the distribution of phospholipid classes, or the amount of cholesterol in the retinas of the monkeys fed the atherogenic diet. These results indicate that the single type of lipid alteration produced in the retina by a diet enriched in saturated fat and cholesterol is a decrease in the polyunsaturation of the retinal phospholipids. The reduction in retinal 22 : 6 content might have significance for photoreceptor function.

Keyword: hyperlipedemia

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro diabetes research.

The interplay between hyper-glycemia and -lipidemia in diabetes mellitus (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for lipid transport to explore the effects of . We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: hyperlipedemia

Increased platelet-fibrinogen interaction in patients with hypercholesterolemia and hypertriglyceridemia.

Binding of fibrinogen to platelets washed from the blood of patients with hypercholesterolemia and hypertriglyceridemia (n = 25) and control donors (n = 12) was compared. In addition, the content of platelet glycoprotein IIb was determined by radioimmunoassay. Fibrinogen was bound in significantly higher amounts (P < 0.02) to hyperlipidaemic platelets activated by ADP than to control ones (107,112 +/- 16,371 and 45,612 +/- 6495 molecules per platelet, respectively). The mean content of GPIIb was the same in hyperlipidaemic and in control platelets (2.06 +/- 0.16 and 1.94 +/- 0.21 micrograms/10(8) platelets, respectively). The amount of fibrinogen bound to the activated hyperlipidaemic platelets showed a positive correlation with total plasma cholesterol and LDL (r = 0.45 and 0.47, respectively) whereas a negative correlation with plasma HDL was found (r = -0.50). The increased expression of fibrinogen binding sites similar to that of hyperlipidaemic platelets could be produced by preincubation of normal platelets with . This was evidenced by a significant increase of fibrinogen binding sites in control platelets. This suggests that either palmitoylation of the receptor or microenvironment changes in the membrane lipid bilayer may be responsible for the enhanced platelet receptor capacity to bind fibrinogen.

Keyword: hyperlipedemia

The CCL5/CCR5 Axis Promotes Vascular Smooth Muscle Cell Proliferation and Atherogenic Phenotype Switching.

induces dysfunction in the smooth muscle cells (SMCs) of the blood vessels, and the vascular remodeling that ensues is a key proatherogenic factor contributing to cardiovascular events. Chemokines and chemokine receptors play crucial roles in vascular remodeling. Here, we examined whether the -derived chemokine CCL5 and its receptor CCR5 influence vascular SMC proliferation, phenotypic switching, and explored the underlying mechanisms.Thoracoabdominal aorta were isolated from wild-type, CCL5 and CCR5 double-knockout mice (CCL5-/-CCR5-/-) fed a high-fat diet (HFD) for 12 weeks. Expression of the contractile, synthetic, and proliferation markers were assayed using immunohistochemical and western blotting. The effects of CCL5 and on cultured SMC proliferation and phenotypic modulation were evaluated using flow cytometry, bromodeoxyuridine (BrdU), and western blotting.Wild-type mice fed an HFD showed markedly increased total cholesterol, triglyceride, and CCL5 serum levels, as well as significantly increased CCL5 and CCR5 expression in the thoracoabdominal aorta vs. normal-diet-fed controls. HFD-fed CCL5-/-CCR5-/- mice showed significantly decreased expression of the synthetic phenotype marker osteopontin and the proliferation marker proliferating cell nuclear antigen, and increased expression of the contractile phenotype marker smooth muscle α-actin in the thoracoabdominal aorta vs. wild-type HFD-fed mice. Human aorta-derived SMCs stimulated with showed significantly increased expression of CCL5, CCR5, and synthetic phenotype markers, as well as increased proliferation. CCL5-treated SMCs showed increased cell cycle regulatory protein expression, paralleling increased synthetic and decreased contractile phenotype marker expression. Inhibition of CCR5 activity by the specific antagonist maraviroc or its expression using small interfering RNA significantly inhibited human aortic SMC proliferation and synthetic phenotype formation. Therefore, CCL5 induces SMC proliferation and phenotypic switching from a contractile to synthetic phenotype via CCR5. CCL5-mediated SMC stimulation activated ERK1/2, Akt/p70S6K, p38 MAPK, and NF-κB signaling. NF-κB inhibition significantly reduced CCR5 expression along with CCR5-induced SMC proliferation and synthetic phenotype formation.-induced CCL5/CCR5 axis activation serves as a pivotal mediator of vascular remodeling, indicating that CCL5 and CCR5 are key chemokine-related factors in atherogenesis. SMC proliferation and synthetic phenotype transformation attenuation by CCR5 pharmacological inhibition may offer a new approach to treatment or prevention of atherosclerotic diseases associated with .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: hyperlipedemia

Effect of Sargassum polycystum (Phaeophyceae)-sulphated polysaccharide extract against acetaminophen-induced during toxic hepatitis in experimental rats.

The effect of Sargassum polycystum crude extract on lipid metabolism was examined against acetaminophen-induced (800 mg/kg body wt., intraperitoneally) during toxic hepatitis in experimental rats. The animals intoxicated with acetaminophen showed significant elevation in the levels of cholesterol, triglycerides and free fatty in both serum and liver tissue. The levels of tissue total lipids and serum LDL-cholesterol were also elevated with depleted levels of serum HDL-cholesterol and tissue phospholipid. The acetaminophen-induced animals showed significant alterations in the activities of lipid metabolizing enzymes serum lecithin cholesterol acyl transferase (LCAT) and hepatic triglyceride lipase (HTGL). The levels of liver tissue fatty acids (saturated, mono and polyunsaturated) such as , stearic , oleic , linoleic , arachidonic and linolenic monitored by gas chromatography were considerably altered in acetaminophen intoxicated animals when compared with control animals. The prior oral administration of Sargassum polycystum (200 mg/kg body wt./day for a period of 15 days) crude extract showed considerable prevention in the severe disturbances of lipid profile and metabolizing enzymes triggered by acetaminophen during hepatic injury. Liver histology also showed convincing supportive evidence regarding their protective nature against fatty changes induced during acetaminophen intoxication. Thus the present study indicates that the protective nature of Sargassum polycystum extract may be due to the presence of active compounds possessing antilipemic property against acetaminophen challenge.(Mol Cell Biochem 276: 89-96, 2005).

Keyword: hyperlipedemia

[Fibrate-induced changes in the serum lipid contents of individual C16 and C18 fatty acids in patients with ].

Keyword: hyperlipedemia

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, , hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid metabolism and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

Keyword: hyperlipedemia

Effect of probucol therapy on plasma fatty composition during postprandial lipemia in hypertriglyceridemia.

To determine the effect of probucol (PR) on fatty composition of plasma lipids in patients with hypertriglyceridemia during the fasting and postprandial states.Open-label, single-center, 6-week treatment, baseline-controlled trial.Outpatient clinical research center.Six patients with established hypertriglyceridemia and no complicating medical conditions.Step 1 American Heart Association diet and no lipid-lowering medications for at least 4 weeks. Lead to a baseline standarized meal ingestion with postprandial blood samplings. After 6 weeks of treatment with 500-mg of PR twice daily and diet, the meal tests were repeated.The clinical status, 3-day food records, postprandial blood samplings (0, 2, 4, 6, and 8 hours) for lipid and fatty acids in whole plasma, triglyceride (TG), phospholipid (PL), and cholesteryl ester (CE) fractions.PR had the following effects: (1) In plasma, cholesterol, high-density lipoprotein cholesterol, apolipoprotein (Apo) A1, and ApoB were decreased postprandially. ApoC111 ratio (heparin-treated plasma versus precipitate) was decreased in the fasting state. (2) The saturated/unsaturated (S/U) fatty ratios in the PL, TG, and CE fractions were elevated postprandially and increased in CE in the fasting state. In the PL fraction it was due to an increase in the percentage of myristic, , stearic, and oleic acids and a decrease in linolenic, eicosatrienoic, and arachidonic acids. In the TG and CE fractions the changes were not due to any particular patterns of fatty acids. The ratio of arachidonic/eicosatrienoic was decreased postprandially in PL and CE fractions. The ratio of eicosatrienoic/linoleic was decreased in PL and increased in CE fractions. (3) The S/U ratio in the lipoprotein (Lp) B and LP(a) lipid components decreased in PL and CE. Lp(a) was more saturated with respect to fatty acids than LpB.PR treatment for 6 weeks increased postprandial S/U fatty ratios. This was due to a combination of an increase in saturated and monounsaturated levels and a decrease in polyunsaturated levels. The effect was most notable in the PL fraction. In those systems dependent on the pattern of fatty acids, the fatty compositional change could modify biological responses in clinically important ways during lipid-lowering therapy. The change toward saturation of fatty acids in the postprandial state may contribute to the antioxidant properties of probucol.

Keyword: hyperlipedemia

Comparison of the effects of medium-chain triacylglycerols, palm oil, and high oleic sunflower oil on plasma triacylglycerol fatty acids and lipid and lipoprotein concentrations in humans.

Although medium-chain triacylglycerols (MCTs, composed of medium-chain fatty acids 8:0 and 10:0) have long been described as having neutral effects on serum cholesterol concentrations, experimental evidence supporting this claim is limited. In a randomized, crossover, metabolic-ward study, we compared the lipid effects of a natural food diet supplemented with either MCTs, palm oil, or high oleic sunflower oil in nine middle-aged men with mild hypercholesterolemia. Rather than having a neutral effect, MCT oil produced total cholesterol concentrations that were not significantly different from those produced by palm oil (MCT oil: 5.87 +/- 0.75 mmol/L; palm oil: 5.79 +/- 0.72 mmol/L) but significantly higher than that produced by high oleic sunflower oil (5.22 +/- 0.52 mmol/L). Low-density-lipoprotein (LDL)-cholesterol concentrations paralleled those of total cholesterol. MCT oil tended to result in higher triacylglycerol concentrations than either palm oil or high oleic sunflower oil, but this difference was not significant. There were no differences in high-density-lipoprotein cholesterol concentrations. The and total saturated fatty content of plasma triacylglycerols in the MCT-oil diet was not significantly different from that in the palm oil diet. On the basis of percentage of energy, this study suggests that medium-chain fatty acids have one-half the potency that has at raising total and LDL-cholesterol concentrations.

Keyword: hyperlipedemia

CD36 Mediated Fatty -Induced Podocyte Apoptosis via Oxidative Stress.

-induced apoptosis mediated by fatty translocase CD36 is associated with increased uptake of ox-LDL or fatty in macrophages, hepatocytes and proximal tubular epithelial cells, leading to atherosclerosis, liver damage and fibrosis in obese patients, and diabetic nephropathy (DN), respectively. However, the specific role of CD36 in podocyte apoptosis in DN with remains poorly investigated.The expression of CD36 was measured in paraffin-embedded kidney tissue samples (Ctr = 18, DN = 20) by immunohistochemistry and immunofluorescence staining. We cultured conditionally immortalized mouse podocytes (MPC5) and treated cells with , and measured CD36 expression by real-time PCR, Western blot analysis and immunofluorescence; lipid uptake by Oil red O staining and BODIPY staining; apoptosis by flow cytometry assay, TUNEL assay and Western blot analysis; and ROS production by DCFH-DA fluorescence staining. All statistical analyses were performed using SPSS 21.0 statistical software.CD36 expression was increased in kidney tissue from DN patients with . upregulated CD36 expression and promoted its translocation from cytoplasm to plasma membrane in podocytes. Furthermore, increased lipid uptake, ROS production and apoptosis in podocytes, Sulfo-N-succinimidyloleate (SSO), the specific inhibitor of the fatty binding site on CD36, decreased -induced fatty accumulation, ROS production, and apoptosis in podocytes. Antioxidant 4-hydroxy-2,2,6,6- tetramethylpiperidine -1-oxyl (tempol) inhibited the overproduction of ROS and apoptosis in podocytes induced by .CD36 mediated fatty -induced podocyte apoptosis via oxidative stress might participate in the process of DN.

Keyword: hyperlipedemia

Free fatty acids increase PGC-1alpha expression in isolated rat islets.

PGC-1alpha mRNA and protein are elevated in islets from multiple animal models of diabetes. Overexpression of PGC-1alpha impairs glucose-stimulated insulin secretion (GSIS). However, it is not well known which metabolic events lead to upregulation of PGC-1alpha in the beta-cells under pathophysiological condition. In present study, we have investigated effects of chronic and hyperglycemia on PGC-1alpha mRNA expression in isolated rat islets. Isolated rat islets are chronically incubated with 0, 0.2 and 0.4 mM oleic / (free fatty acids, FFA) or 5.5 and 25 mM glucose for 72 h. FFA dose-dependently increases PGC-1alpha mRNA expression level in isolated islets. FFA also increases PGC-1alpha expression in mouse beta-cell-derived beta TC3 cell line. In contrast, 25 mM glucose decreases expression level of PGC-1alpha. Inhibition of PGC-1alpha by siRNA improves FFA-induced impairment of GSIS in islets. These data suggest that and hyperglycemia regulate PGC-1alpha expression in islets differently, and elevated PGC-1alpha by FFA plays an important role in chronic -induced beta-cell dysfunction.

Keyword: hyperlipedemia

Palmitoylethanolamide Promotes a Proresolving Macrophage Phenotype and Attenuates Atherosclerotic Plaque Formation.

Objective- Palmitoylethanolamide is an endogenous fatty mediator that is synthetized from membrane phospholipids by N-acyl phosphatidylethanolamine phospholipase D. Its biological actions are primarily mediated by PPAR-α (peroxisome proliferator-activated receptors α) and the orphan receptor GPR55. Palmitoylethanolamide exerts potent anti-inflammatory actions but its physiological role and promise as a therapeutic agent in chronic arterial inflammation, such as atherosclerosis remain unexplored. Approach and Results- First, the polarization of mouse primary macrophages towards a proinflammatory phenotype was found to reduce N-acyl phosphatidylethanolamine phospholipase D expression and palmitoylethanolamide bioavailability. N-acyl phosphatidylethanolamine phospholipase D expression was progressively downregulated in the aorta of apolipoprotein E deficient (ApoE) mice during atherogenesis. N-acyl phosphatidylethanolamine phospholipase D mRNA levels were also downregulated in unstable human plaques and they positively associated with smooth muscle cell markers and negatively with macrophage markers. Second, ApoE mice were fed a high-fat diet for 4 or 16 weeks and treated with either vehicle or palmitoylethanolamide (3 mg/kg per day, 4 weeks) to study the effects of palmitoylethanolamide on early established and pre-established atherosclerosis. Palmitoylethanolamide treatment reduced plaque size in early atherosclerosis, whereas in pre-established atherosclerosis, palmitoylethanolamide promoted signs of plaque stability as evidenced by reduced macrophage accumulation and necrotic core size, increased collagen deposition and downregulation of M1-type macrophage markers. Mechanistically, we found that palmitoylethanolamide, by activating GPR55, increases the expression of the phagocytosis receptor MerTK (proto-oncogene tyrosine-protein kinase MER) and enhances macrophage efferocytosis, indicative of proresolving properties. Conclusions- The present study demonstrates that palmitoylethanolamide protects against atherosclerosis by promoting an anti-inflammatory and proresolving phenotype of lesional macrophages, representing a new therapeutic approach to resolve arterial inflammation.

Keyword: immunity

Comparative compositional analysis of transgenic potato resistant to potato tuber moth (PTM) and its non-transformed counterpart.

In this study, the compositions of transgenic potatoes (TPs) resistant to potato tuber moth (Phthorimaea operculella) were compared with those of its non-transgenic (NTP) counterparts. The light inducible promoter, phosphoenolpyruvate carboxylase led to the expression of Cry1Ab only in the leaves and light-treated tubers of the TPs. No significant differences were found in the moisture, ash, dry weight, total soluble protein, carbohydrate, starch, fiber, ascorbate, cations, anions, fatty acids, and glycoalkaloids contents of TP and NTP. Moreover, light treatment significantly affected the contents of ascorbate, acetate and nitrite anions, , stearic and linolenic fatty acids, α-haconine and α-solanine glycoalkaloids in TP and NTP tubers. While, significant differences were observed in the amino contents in light-treated tubers of TPs than the NTP ones. Although, light treatment in potato tubers resulted in marked metabolic changes, all the variations observed in the metabolites compositions were found to be within the desired reference ranges for potato plants. In conclusion, the results indicated that the TPs were substantially and nutritionally equivalent to the NTP counterparts.

Keyword: immunity

Saturated induces myocardial inflammatory injuries through direct binding to TLR4 accessory protein MD2.

Obesity increases the risk for a number of diseases including cardiovascular diseases and type 2 diabetes. Excess saturated fatty acids (SFAs) in obesity play a significant role in cardiovascular diseases by activating innate responses. However, the mechanisms by which SFAs activate the innate immune system are not fully known. Here we report that (PA), the most abundant circulating SFA, induces myocardial inflammatory injury through the Toll-like receptor 4 (TLR4) accessory protein MD2 in mouse and cell culture experimental models. Md2 knockout mice are protected against PA- and high-fat diet-induced myocardial injury. Studies of cell surface binding, cell-free protein-protein interactions and molecular docking simulations indicate that PA directly binds to MD2, supporting a mechanism by which PA activates TLR4 and downstream inflammatory responses. We conclude that PA is a crucial contributor to obesity-associated myocardial injury, which is likely regulated via its direct binding to MD2.

Keyword: immunity

[The individual fatty acids of blood plasma: biological role of substrates, parameters of quantity and quality, diagnostic of atherosclerosis and atheromotosis.]

The atherosclerosis and atheromotosis are supposed to be, according to phylogenetic theory of general pathology, two etiologically different aphysiological processes, unified by community of pathogenesis. The atherosclerosis is a derangement of biological function of trophology (feeding), biological reaction of exotrophy (external feeding) and biological function of adaptation, biological reaction of compensation in response to deficiency of ῳ-3 and ῳ-6 polyenoic fatty acids. In case of deficiency of polyenoic fatty acids in cells and during synthesis of eicosanoids of group I from unsaturated endogenous ῳ-6 С20: 3 digomo-γ-linoleic unsaturated fatty , atherosclerosis is developed, a complex metabolism disorder in vivo. The atheromotosis is a derangement of biological function of endoecology, biological reactions of inflammation and inherent . This incomplete utilization in intima of arteries of non-ligand lipoproteins of very low → low density under effect not of polyfunctional resident macrophage but monocytes of hematogenic origin without expression of hydrolase of polyenoic ethers of cholesterol. In intima, in area of cumulation of endogenous phlogogens (initiator of inflammation) from the pool of intra-vascular medium, polyenoic unsaturated fatty acids are cumulated that were not absorbed by cells in structure of ligand low density lipoproteins using apoB-100- endocytosis. The pathogenic factor of atherosclerosis - derangement of biological function of trophology. biological function of exotrophy under alimentary deficiency of in vivo of ῳ-3 and ῳ-6 polyenoic fatty acids with physiological parameters of feeding. The pathogenic factor of atheromotosis - phylogenetically herbivorous (carnivorous) human misusing of animal (meat) food, unsaturated fatty acids, development by hepatocytes of a large number of triglycerides and lipoproteins of very low density of the same name. The late in phylogenesis insulin-dependent lipoproteins of very low density transfer lipoproteins of very low density to cells slowly. The cells absorb them also slowly. The cumulation of non-ligand lipoproteins of very low density → low density in blood competitively blocks physiological absorption of polyenoic unsaturated fatty acids by cells in structure of physiological lipoproteins of low density. The atherosclerosis occurs blood flow and atheromotosis in intima of arteries of elastic type.

Keyword: immunity

Eicosanoids mediate Galleria mellonella immune response to hemocoel injection of entomopathogenic nematode cuticles.

Entomopathogenic nematodes are symbiotically associated with bacteria and widely used in biological control of insect pests. The interference of symbiotic bacteria with insect host immune responses is fairly well documented. However, knowledge of mechanisms regulating parasite–host interactions still remains fragmentary. In this study, we used nematode (Steinernema carpocapsae and Heterorhabditis bacteriophora) cuticles and Galleria mellonella larvae as parasite–host model, focused on the changes of innate immune parameters of the host in the early phase of nematode cuticle infection and investigated the role of eicosanoid biosynthesis pathway in the process. The results showed that injection of either S. carpocapsae or H. bacteriophora cuticles into the larval hemocoel both resulted in significant decreases in the key innate immune parameters (e.g., hemocyte density, microaggregation, phagocytosis and encapsulation abilities of hemocyte, and phenoloxidase and antibacterial activities of the cell-free hemolymph). Our study indicated that the parasite cuticles could actively suppress the innate immune response of the G. mellonella host. We also found that treating G. mellonella larvae with dexamethasone and indomethacin induced similar depression in the key innate immune parameters to the nematode cuticles. However, these effects were reversed when dexamethasone, indomethacin, or nematode cuticles were injected together with arachidonic . Additionally, we found that did not reverse the influence of the dexamethasone, indomethacin, or nematode cuticles on the innate immune responses. Therefore, we inferred from our results that both S. carpocapsae and H. bacteriophora cuticles inhibited eicosanoid biosynthesis to induce host immunodepression.

Keyword: immunity

Fatty metabolism in the host and commensal bacteria for the control of intestinal immune responses and diseases.

Intestinal tissue has a specialized immune system that exhibits an exquisite balance between active and suppressive responses important for the maintenance of health. Intestinal is functionally affected by both diet and gut commensal bacteria. Here, we review the effects of fatty acids on the regulation of intestinal and immunological diseases, revealing that dietary fatty acids and their metabolites play an important role in the regulation of allergy, inflammation, and immunosurveillance in the intestine. Several lines of evidence have revealed that some dietary fatty acids are converted to biologically active metabolites by enzymes not only in the host but also in the commensal bacteria. Thus, biological interaction between diet and commensal bacteria could form the basis of a new era in the control of host and its associated diseases.

Keyword: immunity

Branched Fatty Esters of Hydroxy Fatty Acids (FAHFAs) Protect against Colitis by Regulating Gut Innate and Adaptive Immune Responses.

We recently discovered a structurally novel class of endogenous lipids, branched esters of hydroxy stearic acids (PAHSAs), with beneficial metabolic and anti-inflammatory effects. We tested whether PAHSAs protect against colitis, which is a chronic inflammatory disease driven predominantly by defects in the innate mucosal barrier and adaptive immune system. There is an unmet clinical need for safe and well tolerated oral therapeutics with direct anti-inflammatory effects. Wild-type mice were pretreated orally with vehicle or 5-PAHSA (10 mg/kg) and 9-PAHSA (5 mg/kg) once daily for 3 days, followed by 10 days of either 0% or 2% dextran sulfate sodium water with continued vehicle or PAHSA treatment. The colon was collected for histopathology, gene expression, and flow cytometry. Intestinal crypt fractions were prepared for ex vivo bactericidal assays. Bone marrow-derived dendritic cells pretreated with vehicle or PAHSA and splenic CD4 T cells from syngeneic mice were co-cultured to assess antigen presentation and T cell activation in response to LPS. PAHSA treatment prevented weight loss, improved colitis scores (stool consistency, hematochezia, and mouse appearance), and augmented intestinal crypt Paneth cell bactericidal potency via a mechanism that may involve GPR120. In vitro, PAHSAs attenuated dendritic cell activation and subsequent T cell proliferation and Th1 polarization. The anti-inflammatory effects of PAHSAs in vivo resulted in reduced colonic T cell activation and pro-inflammatory cytokine and chemokine expression. These anti-inflammatory effects appear to be partially GPR120-dependent. We conclude that PAHSA treatment regulates innate and adaptive immune responses to prevent mucosal damage and protect against colitis. Thus, PAHSAs may be a novel treatment for colitis and related inflammation-driven diseases.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: immunity

Frontline Science: Specialized proresolving lipid mediators inhibit the priming and activation of the macrophage NLRP3 inflammasome.

The prototypic proinflammatory cytokine IL-1β plays a central role in innate immunity and inflammatory disorders. The formation of mature IL-1β from an inactive pro-IL-1β precursor is produced via nonconventional multiprotein complexes called the inflammasomes, of which the most common is the nucleotide-binding domain leucine-rich repeat-containing protein 3 (NLRP3) inflammasome composed by NLRP3, (ASC) apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (CARD), and caspase-1. Specialized proresolving mediators (SPMs) promote resolution of , which is an essential process to maintain host health. SPMs prevent excessive by terminating the inflammatory response and returning to tissue homeostasis without immunosupression. This study tested the hypothesis that modulation of the NLRP3 inflammasome in macrophages is one mechanism involved in the SPM-regulated processes during resolution. Our findings demonstrate that the SPM resolvin D2 (RvD2) suppressed the expression of pro-IL-1β and reduced the secretion of mature IL-1β in bone marrow-derived macrophages challenged with LPS+ATP (classical NLRP3 inflammasome model) or LPS+palmitate (lipotoxic model). Similar findings were observed in thioglycolate-elicited peritoneal macrophages, in which RvD2 remarkably reduced ASC oligomerization, inflammasome assembly, and caspase-1 activity. In vivo, in a self-resolving zymosan A-induced peritonitis model, RvD2 blocked the NLRP3 inflammasome leading to reduced release of IL-1β into the exudates, repression of osteopontin, and MCP-1 expression and induction of M2 markers of resolution (i.e., CD206 and arginase-1) in peritoneal macrophages. RvD2 inhibitory actions were receptor mediated and were abrogated by a selective GPR18 antagonist. Together, these findings support the hypothesis that SPMs have the ability to inhibit the priming and to expedite the deactivation of the NLRP3 inflammasome in macrophages during the resolution process.©2018 Society for Leukocyte Biology.

Keyword: immunity

An unexpected link between fatty synthase and cholesterol synthesis in proinflammatory macrophage activation.

Different immune activation states require distinct metabolic features and activities in immune cells. For instance, inhibition of fatty synthase (FASN), which catalyzes the synthesis of long-chain fatty acids, prevents the proinflammatory response in macrophages; however, the precise role of this enzyme in this response remains poorly defined. Consistent with previous studies, we found here that FASN is essential for lipopolysaccharide-induced, Toll-like receptor (TLR)-mediated macrophage activation. Interestingly, only agents that block FASN upstream of acetoacetyl-CoA synthesis, including the well-characterized FASN inhibitor C75, inhibited TLR4 signaling, while those acting downstream had no effect. We found that acetoacetyl-CoA could overcome C75\'s inhibitory effect, whereas other FASN metabolites, including palmitate, did not prevent C75-mediated inhibition. This suggested an unexpected role for acetoacetyl-CoA in that is independent of its role in palmitate synthesis. Our evidence further suggested that acetoacetyl-CoA arising from FASN activity promotes cholesterol production, indicating a surprising link between fatty synthesis and cholesterol synthesis. We further demonstrate that this process is required for TLR4 to enter lipid rafts and facilitate TLR4 signaling. In conclusion, we have uncovered an unexpected link between FASN and cholesterol synthesis that appears to be required for TLR signal transduction and proinflammatory macrophage activation.© 2018 Carroll et al.

Keyword: immunity

Novel lipopeptides of ESAT-6 induce strong protective against Mycobacterium tuberculosis: Routes of immunization and TLR agonists critically impact vaccine\'s efficacy.

Mycobacterium tuberculosis (Mtb), the bacterial cause of tuberculosis, is a leading infectious agent worldwide. The development of a new vaccine against Mtb is essential to control global spread of tuberculosis, since the current vaccine BCG is not very effective and antibiotic resistance is a serious, burgeoning problem. ESAT-6 is a secreted protein of Mtb, which is absent in BCG but has been implicated in inducing protective against Mtb. Peptide based subunit vaccines are attractive due to their safety and high specificity in eliciting immune responses, but small synthetic peptides are usually not very immunogenic. We have designed a novel subunit vaccine for Mtb by using simple lipid () modified derivatives of peptides from ESAT-6 protein corresponding to dominant human T cell epitopes and examined their ability to stimulate protective against Mtb by intranasal and subcutaneous immunization in mice. We also investigated how individual TLR agonists as adjuvants (PolyI:C, MPL and GDQ) contribute to enhancing the induced immune responses and resulting protective efficacy of our vaccine. We observed that single C-terminal palmitoyl-lysine modified lipopeptides derived from ESAT-6 induce significant cellular immune responses on their own upon mucosal and subcutaneous immunizations. Intriguingly, a combination of immunogenic lipopeptides of ESAT-6 antigen exhibited local (pulmonary) and systemic immune responses along with efficient protective efficacy when administered intranasally or subcutaneously. Surprisingly, combination of ESAT-6 derived lipopeptides with a TLR-4 agonist (MPL) enhanced protection, whereas TLR-3 (Poly I:C) and TLR-7/8 agonists (gardiquimod, GDQ) led to reduced protection associated with specific local and systemic immune modulation. Our studies demonstrate the potential of ESAT-6 derived lipopeptides as a promising vaccine candidate against Mtb, and emphasize that selection of adjuvant is critical for the success of vaccines. These findings demonstrate the promise of synthetic lipopeptides as the basis of a subunit vaccine for TB.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: immunity

Ginsenoside Rb1 improves leptin sensitivity in the prefrontal cortex in obese mice.

Obesity impairs leptin-induced regulation of brain-derived neurotrophic factor (BDNF) expression and synaptogenesis, which has been considered to be associated with the incidence of neuronal degenerative diseases, cognitive decline, and depression. Ginsenoside Rb1 (Rb1), a major bioactive component of ginseng, is known to have an antiobesity effect and improve cognition. This study examined whether Rb1 can improve central leptin effects on BDNF expression and synaptogenesis in the prefrontal cortex during obesity using an in vivo and an in vitro model.Ginsenoside Rb1 (Rb1) chronic treatment improved central leptin sensitivity, leptin-JAK2-STAT3 signaling, and leptin-induced regulation of BDNF expression in the prefrontal cortex of high-fat diet-induced obese mice. In cultured prefrontal cortical neurons, , the saturated fat, impaired leptin-induced BDNF expression, reduced the immunoreactivity and mRNA expression of synaptic proteins, and impaired leptin-induced neurite outgrowth and synaptogenesis. Importantly, Rb1 significantly prevented these pernicious effects induced by .These results indicate that Rb1 reverses central leptin resistance and improves leptin-BDNF-neurite outgrowth and synaptogenesis in the prefrontal cortical neurons. Thus, Rb1 supplementation may be a beneficial avenue to treat obesity-associated neurodegenerative disorders.© 2017 John Wiley & Sons Ltd.

Keyword: immunity

Diabetes Mellitus-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and via Nuclear Mechanisms.

Objective- Macrophages play key roles in and diabetic vascular complications. Emerging evidence implicates long noncoding RNAs in , but their role in macrophage dysfunction associated with inflammatory diabetic complications is unclear and was therefore investigated in this study. Approach and Results- RNA-sequencing and real-time quantitative PCR demonstrated that a long noncoding RNA Dnm3os (dynamin 3 opposite strand) is upregulated in bone marrow-derived macrophages from type 2 diabetic db/db mice, diet-induced insulin-resistant mice, and diabetic ApoE mice, as well as in monocytes from type 2 diabetic patients relative to controls. Diabetic conditions (high glucose and ) induced Dnm3os in mouse and human macrophages. Promoter reporter analysis and chromatin immunoprecipitation assays demonstrated that diabetic conditions induce Dnm3os via NF-κB activation. RNA fluorescence in situ hybridization and real-time quantitative PCRs of subcellular fractions demonstrated nuclear localization and chromatin enrichment of Dnm3os in macrophages. Stable overexpression of Dnm3os in macrophages altered global histone modifications and upregulated and immune response genes and phagocytosis. Conversely, RNAi-mediated knockdown of Dnm3os attenuated these responses. RNA pull-down assays with macrophage nuclear lysates identified nucleolin and ILF-2 (interleukin enhancer-binding factor 2) as protein binding partners of Dnm3os, which was further confirmed by RNA fluorescence in situ hybridization immunofluorescence. Furthermore, nucleolin levels were decreased in diabetic conditions, and its knockdown enhanced Dnm3os-induced inflammatory gene expression and histone H3K9-acetylation at their promoters. Conclusions- These results demonstrate novel mechanisms involving upregulation of long noncoding RNA Dnm3os, disruption of its interaction with nucleolin, and epigenetic modifications at target genes that promote macrophage inflammatory phenotype in diabetes mellitus. The data could lead to long noncoding RNA-based therapies for inflammatory diabetes mellitus complications.

Keyword: immunity

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: immunity

Assessment of free fatty acids and cholesteryl esters delivered in liposomes as novel class of antibiotic.

Healthcare associated infections (HAI) with multidrug-resistant (MDR) bacteria continue to be a global threat, highlighting an urgent need for novel antibiotics. In this study, we assessed the potential of free fatty acids and cholesteryl esters that form part of the innate host defense as novel antibacterial agents for use against MDR bacteria.Liposomes of six different phospholipid mixtures were employed as carrier for six different fatty acids and four different cholesteryl esters. Using a modified MIC assay based on DNA quantification with the fluoroprobe Syto9, formulations were tested against Gram-positive and Gram-negative bacteria implicated in HAI. Formulations with MIC values in the low μg/mL range were further subjected to determination of minimal bactericidal activity, hemolysis assay with sheep erythrocytes, and cytotoxicity testing with the human liver cell line HepG2. The potential for synergistic activity with a standard antibiotic was also probed. and stearic prepared in carrier 4 (PA4 and SA4, respectively) were identified as most active lipids (MIC against MDR Staphylococcus epidermidis was 0.5 and 0.25\xa0μg/mL, respectively; MIC against vancomycin resistant Enterococcus faecalis (VRE) was 2 and 0.5\xa0μg/mL, respectively). Cholesteryl linoleate formulated with carrier 3 (CL3) exhibited activity against the S. epidermidis strain (MIC 1\xa0μg/mL) and a Pseudomonas aeruginosa strain (MIC 8\xa0μg/mL) and lowered the vancomycin MIC for VRE from 32-64\xa0μg/mL to as low as 4\xa0μg/mL. At 90\xa0μg/mL PA4, SA4, and CL3 effected less than 5\xa0% hemolysis over 3\xa0h and PA4 and CL3 did not exhibit significant cytotoxic activity against HepG2 cells when applied at 100\xa0μg/mL over 48\xa0h.Our results showed that selected fatty acids and cholesteryl esters packaged with phospholipids exhibit antibacterial activity against Gram-positive and Gram-negative bacteria and may augment the activity of antibiotics. Bactericidal activity could be unlinked from hemolytic and cytotoxic activity and the type of phospholipid carrier greatly influenced the activity. Thus, fatty acids and cholesteryl esters packaged in liposomes may have potential as novel lipophilic antimicrobial agents.

Keyword: immunity

Free fatty acids sensitize dendritic cells to amplify TH1/TH17-immune responses.

Obesity is associated with body fat gain and impaired glucose metabolism. Here, we identified both body fat gain in obesity and impaired glucose metabolism as two independent risk factors for increased serum levels of free fatty acids (FFAs). Since obesity is associated with increased and/or delayed resolution of observed in various chronic inflammatory diseases such as psoriasis, we investigated the impact of FFAs on human monocyte-derived and mouse bone marrow-derived dendritic cell (DCs) functions relevant for the pathogenesis of chronic . FFAs such as (PA) and oleic (OA) did not affect the pro-inflammatory immune response of DCs. In contrast, PA and OA sensitize DCs resulting in augmented secretion of TH1/TH17-instructive cytokines upon pro-inflammatory stimulation. Interestingly, obesity in mice worsened a TH1/TH17-driven psoriasis-like skin . Strong correlation of the amount of total FFA, PA, and OA in serum with the severity of skin points to a critical role of FFA in obesity-mediated exacerbation of skin . Our data suggest that increased levels of FFAs might be a predisposing factor promoting a TH1/TH17-mediated such as psoriasis in response to an inflammatory danger signal.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: immunity

Palmitoylethanolamide induces microglia changes associated with increased migration and phagocytic activity: involvement of the CB2 receptor.

The endogenous fatty amide palmitoylethanolamide (PEA) has been shown to exert anti-inflammatory actions mainly through inhibition of the release of pro-inflammatory molecules from mast cells, monocytes and macrophages. Indirect activation of the endocannabinoid (eCB) system is among the several mechanisms of action that have been proposed to underlie the different effects of PEA in vivo. In this study, we used cultured rat microglia and human macrophages to evaluate whether PEA affects eCB signaling. PEA was found to increase CB2 mRNA and protein expression through peroxisome proliferator-activated receptor-α (PPAR-α) activation. This novel gene regulation mechanism was demonstrated through: (i) pharmacological PPAR-α manipulation, (ii) PPAR-α mRNA silencing, (iii) chromatin immunoprecipitation. Moreover, exposure to PEA induced morphological changes associated with a reactive microglial phenotype, including increased phagocytosis and migratory activity. Our findings suggest indirect regulation of microglial CB2R expression as a new possible mechanism underlying the effects of PEA. PEA can be explored as a useful tool for preventing/treating the symptoms associated with neuroinflammation in CNS disorders.

Keyword: immunity

Dipeptidyl peptidase 9 enzymatic activity influences the expression of neonatal metabolic genes.

The success of dipeptidyl peptidase 4 (DPP4) inhibition as a type 2 diabetes therapy has encouraged deeper examination of the post-proline DPP enzymes. DPP9 has been implicated in immunoregulation, disease pathogenesis and metabolism. The DPP9 enzyme-inactive (Dpp9 gene knock-in; Dpp9 gki) mouse displays neonatal lethality, suggesting that DPP9 enzyme activity is essential in neonatal development. Here we present gene expression patterns in these Dpp9 gki neonatal mice. Taqman PCR arrays and sequential qPCR assays on neonatal liver and gut revealed differential expression of genes involved in cell growth, innate and metabolic pathways including long-chain-fatty- uptake and esterification, long-chain fatty acyl-CoA binding, trafficking and transport into mitochondria, lipoprotein metabolism, adipokine transport and gluconeogenesis in the Dpp9 gki mice compared to wild type. In a liver cell line, Dpp9 knockdown increased AMP-activated protein kinase phosphorylation, which suggests a potential mechanism. DPP9 protein levels in liver cells were altered by treatment with EGF, HGF, insulin or palmitate, suggesting potential natural DPP9 regulators. These gene expression analyses of a mouse strain deficient in DPP9 enzyme activity show, for the first time, that DPP9 enzyme activity regulates metabolic pathways in neonatal liver and gut.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: immunity

TLR4 knockout attenuated high fat diet-induced cardiac dysfunction via NF-κB/JNK-dependent activation of autophagy.

Obesity is commonly associated with a low grade systemic inflammation, which may contribute to the onset and development of myocardial remodeling and contractile dysfunction. Toll-like receptor 4 (TLR4) plays an important role in innate and inflammation although its role in high fat diet-induced obesity cardiac dysfunction remains elusive. This study was designed to examine the effect of TLR4 ablation on high fat diet intake-induced cardiac anomalies, if any, and underlying mechanism(s) involved. Wild-type (WT) and TLR4 knockout mice were fed normal or high fat (60% calorie from fat) diet for 12weeks prior to assessment of mechanical and intracellular Ca properties. The inflammatory signaling proteins (TLR4, NF-κB, and JNK) and autophagic markers (Atg5, Atg12, LC3B and p62) were evaluated. Our results revealed that high fat diet intake promoted obesity, marked decrease in fractional shortening, and cardiomyocyte contractile capacity with dampened intracellular Ca release and clearance, elevated ROS generation and oxidative stress as measured by aconitase activity, the effects of which were significantly attenuated by TLR4 knockout. In addition, high fat intake downregulated levels of Atg5, Atg12 and LC3B, while increasing p62 accumulation. TLR4 knockout itself did not affect Atg5, Atg12, LC3B and p62 levels while it reconciled high fat diet intake-induced changes in autophagy. In addition, TLR4 knockout alleviated high fat diet-induced phosphorylation of IKKβ, JNK and mTOR. In vitro study revealed that suppressed cardiomyocyte contractile function, the effect of which was inhibited the TLR4 inhibitor CLI-095, the JNK inhibitor AS601245 or the NF-κB inhibitor Celastrol. Taken together, these data showed that TLR4 knockout ameliorated high fat diet-induced cardiac contractile and intracellular Ca anomalies through inhibition of inflammation and ROS, possibly through a NF-κB/JNK-dependent activation of autophagy. This article is part of a Special Issue entitled: Genetic and epigenetic control of heart failure - edited by Jun Ren & Megan Yingmei Zhang.Copyright © 2017. Published by Elsevier B.V.

Keyword: immunity

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: immunity

Palmitate differentially regulates the polarization of differentiating and differentiated macrophages.

The tissue accumulation of M1 macrophages in patients with metabolic diseases such as obesity and type 2 diabetes mellitus has been well-documented. Interestingly, it is an accumulation of M2 macrophages that is observed in the adipose, liver and lung tissues, as well as in the circulation, of patients who have had major traumas such as a burn injury or sepsis; however, the trigger for the M2 polarization observed in these patients has not yet been identified. In the current study, we explored the effects of chronic palmitate and high glucose treatment on macrophage differentiation and function in murine bone-marrow-derived macrophages. We found that chronic treatment with palmitate decreased phagocytosis and HLA-DR expression in addition to inhibiting the production of pro-inflammatory cytokines. Chronic palmitate treatment of bone marrows also led to M2 polarization, which correlated with the activation of the peroxisome proliferator-activated receptor-γ signalling pathway. Furthermore, we found that chronic palmitate treatment increased the expression of multiple endoplasmic reticulum (ER) stress markers, including binding immunoglobulin protein. Preconditioning with the universal ER stress inhibitor 4-phenylbutyrate attenuated ER stress signalling and neutralized the effect of palmitate, inducing a pro-inflammatory phenotype. We confirmed these results in differentiating human macrophages, showing an anti-inflammatory response to chronic palmitate exposure. Though alone it did not promote M2 polarization, hyperglycaemia exacerbated the effects of palmitate. These findings suggest that the dominant accumulation of M2 in adipose tissue and liver in patients with critical illness may be a result of hyperlipidaemia and hyperglycaemia, both components of the hypermetabolism observed in critically ill patients.© 2015 John Wiley & Sons Ltd.

Keyword: immunity

Adipocyte Fatty Binding Protein Potentiates Toxic Lipids-Induced Endoplasmic Reticulum Stress in Macrophages via Inhibition of Janus Kinase 2-dependent Autophagy.

Lipotoxicity is implicated in the pathogenesis of obesity-related inflammatory complications by promoting macrophage infiltration and activation. Endoplasmic reticulum (ER) stress and adipocyte fatty binding protein (A-FABP) play key roles in obesity and mediate inflammatory activity through similar signaling pathways. However, little is known about their interplay in lipid-induced inflammatory responses. Here, we showed that prolonged treatment of (PA) increased ER stress and expression of A-FABP, which was accompanied by reduced autophagic flux in macrophages. Over-expression of A-FABP impaired PA-induced autophagy associating with enhanced ER stress and pro-inflammatory cytokine production, while genetic ablation or pharmacological inhibition of A-FABP reversed the conditions. PA-induced expression of autophagy-related protein (Atg)7 was attenuated in A-FABP over-expressed macrophages, but was elevated in A-FABP-deficient macrophages. Mechanistically, A-FABP potentiated the effects of PA by inhibition of Janus Kinase (JAK)2 activity, thus diminished PA-induced Atg7 expression contributing to impaired autophagy and further augmentation of ER stress. These findings suggest that A-FABP acts as autophagy inhibitor to instigate toxic lipids-induced ER stress through inhibition of JAK2-dependent autophagy, which in turn triggers inflammatory responses in macrophages. A-FABP-JAK2 axis may represent an important pathological pathway contributing to obesity-related inflammatory diseases.

Keyword: immunity

MD2 Blockage Protects Obesity-Induced Vascular Remodeling via Activating AMPK/Nrf2.

Obesity and increased free fatty (FFA) levels are tightly linked with vascular oxidative stress and remodeling. Myeloid differentiation 2 (MD2), an important protein in innate , is requisite for endotoxin lipopolysaccharide responsiveness. This study shows that (PA) also bonds to MD2, initiating cardiac inflammatory injury. However, it is not clear whether MD2 plays a role in noninflammatory systems such as obesity- and FFA-related oxidative stress involved in vascular remodeling and injury. The aim of this study is to examine whether MD2 participates in reactive oxygen species increase and vascular remodeling.Male MD2 mice and wild-type littermates with a C57BL/6 background were fed a high-fat diet (HFD) to establish obesity-induced vascular remodeling. Rat aortic endothelial cells (RAECs) and vascular smooth muscle cells (VSMCs) were treated with PA to induce oxidative stress and injury.In vivo, MD2 deficiency significantly reduced HFD-induced vascular oxidative stress, fibrosis, and remodeling, accompanied with AMP-activated kinase (AMPK) activation and nuclear factor erythroid (Nrf2) upregulation. In VSMCs and RAECs, inhibition of MD2 by neutralizing monoclonal antibody to MD2 or small interfering RNA knockdown significantly activated the AMPK/Nrf2-signaling pathway and reduced PA-induced oxidative stress and cell injury.It was demonstrated that the deletion or inhibition of MD2 protects against HFD/FFA-induced vascular oxidative stress and remodeling by activating the AMPK/Nrf2-signaling pathway.© 2017 The Obesity Society.

Keyword: immunity

Long-term persistence of induced by OVA-coupled gas-filled microbubble vaccination partially protects mice against infection by OVA-expressing Listeria.

Vaccination aims at generating memory immune responses able to protect individuals against pathogenic challenges over long periods of time. Subunit vaccine formulations based on safe, but poorly immunogenic, antigenic entities must be combined with adjuvant molecules to make them efficient against infections. We have previously shown that gas-filled microbubbles (MB) are potent antigen-delivery systems. This study compares the ability of various ovalbumin-associated MB (OVA-MB) formulations to induce antigen-specific memory immune responses and evaluates long-term protection toward bacterial infections. When initially testing dendritic cells reactivity to MB constituents, exhibited the highest degree of activation. Subcutaneous immunization of naïve wild-type mice with the OVA-MB formulation comprising the highest content and devoid of PEG2000 was found to trigger the more pronounced Th1-type response, as reflected by robust IFN-γ and IL-2 production. Both T cell and antibody responses persisted for at least 6 months after immunization. At that time, systemic infection with OVA-expressing Listeria monocytgenes was performed. Partial protection of vaccinated mice was demonstrated by reduction of the bacterial load in both the spleen and liver. We conclude that antigen-bound MB exhibit promising properties as a vaccine candidate ensuring prolonged maintenance of protective .Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: immunity

Lipid Quality in Infant Nutrition: Current Knowledge and Future Opportunities.

Dietary lipids are key for infants to not only meet their high energy needs but also fulfill numerous metabolic and physiological functions critical to their growth, development, and health. The lipid composition of breast milk varies during lactation and according to the mother\'s diet, whereas the lipid composition of infant formulae varies according to the blend of different fat sources. This report compares the compositions of lipids in breast milk and infant formulae, and highlights the roles of dietary lipids in term and preterm infants and their potential biological and health effects. The major differences between breast milk and formulae lie in a variety of saturated fatty acids (such as , including its structural position) and unsaturated fatty acids (including arachidonic and docosahexaenoic ), cholesterol, and complex lipids. The functional outcomes of these differences during infancy and for later child and adult life are still largely unknown, and some of them are discussed, but there is consensus that opportunities exist for improvements in the qualitative lipid supply to infants through the mother\'s diet or infant formulae. Furthermore, research is required in several areas, including the needs of term and preterm infants for long-chain polyunsaturated fatty acids, the sites of action and clinical effects of lipid mediators on and inflammation, the role of lipids on metabolic, neurological, and immunological outcomes, and the mechanisms by which lipids act on short- and long-term health.

Keyword: immunity

TRIF-dependent Toll-like receptor signaling suppresses transcription in hepatocytes and prevents diet-induced hepatic steatosis.

Nonalcoholic fatty liver disease (NAFLD) includes a spectrum of diseases that ranges in severity from hepatic steatosis to steatohepatitis, the latter of which is a major predisposing factor for liver cirrhosis and cancer. Toll-like receptor (TLR) signaling, which is critical for innate , is generally believed to aggravate disease progression by inducing inflammation. Unexpectedly, we found that deficiency in TIR domain-containing adaptor-inducing interferon-β (TRIF), a cytosolic adaptor that transduces some TLR signals, worsened hepatic steatosis induced by a high-fat diet (HFD) and that such exacerbation was independent of myeloid cells. The aggravated steatosis in mice was due to the increased hepatocyte transcription of the gene encoding stearoyl-coenzyme A (CoA) desaturase 1 (SCD1), the rate-limiting enzyme for lipogenesis. Activation of the TRIF pathway by polyinosinic:polycytidylic [poly(I:C)] suppressed the increase in SCD1 abundance induced by or an HFD and subsequently prevented lipid accumulation in hepatocytes. Interferon regulatory factor 3 (IRF3), a transcriptional regulator downstream of TRIF, acted as a transcriptional suppressor by directly binding to the promoter. These results suggest an unconventional metabolic function for TLR/TRIF signaling that should be taken into consideration when seeking to pharmacologically inhibit this pathway.Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Keyword: immunity

STING-IRF3 Triggers Endothelial Inflammation in Response to Free Fatty -Induced Mitochondrial Damage in Diet-Induced Obesity.

Metabolic stress in obesity induces endothelial inflammation and activation, which initiates adipose tissue inflammation, insulin resistance, and cardiovascular diseases. However, the mechanisms underlying endothelial inflammation induction are not completely understood. Stimulator of interferon genes (STING) is an important molecule in and inflammation. In the present study, we sought to determine the role of STING in -induced endothelial activation/inflammation.In cultured endothelial cells, treatment activated STING, as indicated by its perinuclear translocation and binding to interferon regulatory factor 3 (IRF3), leading to IRF3 phosphorylation and nuclear translocation. The activated IRF3 bound to the promoter of ICAM-1 (intercellular adhesion molecule 1) and induced ICAM-1 expression and monocyte-endothelial cell adhesion. When analyzing the upstream signaling, we found that activated STING by inducing mitochondrial damage. treatment caused mitochondrial damage and leakage of mitochondrial DNA into the cytosol. Through the cytosolic DNA sensor cGAS (cyclic GMP-AMP synthase), the mitochondrial damage and leaked cytosolic mitochondrial DNA activated the STING-IRF3 pathway and increased ICAM-1 expression. In mice with diet-induced obesity, the STING-IRF3 pathway was activated in adipose tissue. However, STING deficiency ( ) partially prevented diet-induced adipose tissue inflammation, obesity, insulin resistance, and glucose intolerance.The mitochondrial damage-cGAS-STING-IRF3 pathway is critically involved in metabolic stress-induced endothelial inflammation. STING may be a potential therapeutic target for preventing cardiovascular diseases and insulin resistance in obese individuals.© 2017 American Heart Association, Inc.

Keyword: immunity

In vitro Modulation of the LPS-Induced Proinflammatory Profile of Hepatocytes and Macrophages- Approaches for Intervention in Obesity?

Low grade endotoxemia is a feature of obesity which is linked to development of steatohepatitis in non-alcoholic fatty liver disease. In this study, macrophages (J774) and hepatocytes (HepG2) were stimulated with lipopolysaccharide (LPS) from E. coli 0111: B4 and analyzed for modulation of this response when preconditioned or stimulated subsequent to LPS, with different doses of Vitamin D3 or docosahexaenoic (DHA) over a time period of 1 and 5 days. Pro-inflammatory TNFα and pro-fibrotic TGFβ released into the supernatants were measured by ELISA; qPCR was performed for Srebp-1c and PPARα mRNA (genes for products involved in fatty synthesis and catabolism, respectively). Vitamin D3 and DHA exerted a consistent, dose dependent anti-inflammatory effect, and increased PPARα relative to Srebp-1c in both cell types. By contrast, addition of free fatty acids (FFA, oleic / 2:1) caused aggravation of LPS-induced inflammatory reaction and an increase of Srebp-1c relative to PPARα. Our results argue in favor of dietary supplementation of Vitamin D3 or DHA (and avoidance of monounsaturated/saturated fatty acids) to alleviate development of fatty liver disease.

Keyword: immunity

WAX INDUCER1 (HvWIN1) transcription factor regulates free fatty biosynthetic genes to reinforce cuticle to resist Fusarium head blight in barley spikelets.

Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most devastating diseases of wheat and barley. Resistance to FHB is highly complex and quantitative in nature, and is most often classified as resistance to spikelet infection and resistance to spread of pathogen through the rachis. In the present study, a resistant (CI9831) and a susceptible (H106-371) two-row barley genotypes, with contrasting levels of spikelet resistance to FHB, pathogen or mock-inoculated, were profiled for metabolites based on liquid chromatography and high resolution mass spectrometry. The key resistance-related (RR) metabolites belonging to fatty acids, phenylpropanoids, flavonoids and terpenoid biosynthetic pathways were identified. The free fatty acids (FFAs) linoleic and acids were among the highest fold change RR induced (RRI) metabolites. These FFAs are deposited as cutin monomers and oligomers to reinforce the cuticle, which acts as a barrier to pathogen entry. Quantitative real-time PCR studies revealed higher expressions of KAS2, CYP86A2, CYP89A2, LACS2 and WAX INDUCER1 (HvWIN1) transcription factor in the pathogen-inoculated resistant genotype than in the susceptible genotype. Knockdown of HvWIN1 by virus-induced genes silencing (VIGS) in resistant genotype upon pathogen inoculation increased the disease severity and fungal biomass, and decreased the abundance of FFAs like linoleic and acids. Notably, the expression of CYP86A2, CYP89A2 and LAC2 genes was also suppressed, proving the link of HvWIN1 in regulating these genes in cuticle biosynthesis as a defense response.© The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Keyword: immunity

Lipids as activators of innate in peptide vaccine delivery.

Innate immune system plays an important role in pathogen detection and the recognition of vaccines, mainly through pattern recognition receptors (PRRs) that identify pathogen components (danger signals). One of the typically recognised bacterial components are lipids in conjugation with peptides, proteins and saccharides. Lipidic compounds are readily recognised by the immune system, and thus are ideal candidates for peptide-based vaccine delivery. Thus, bacterial or synthetic lipids mixed with, or conjugated to, antigens have shown adjuvanting properties. These systems have many advantages over traditional adjuvants, including low toxicity and good efficacy for stimulating mucosal and systemic immune responses.The most recent literature on the role of lipids in stimulation of immune responses was selected for this review. The vast majority of reviewed papers were published in the last decade. Older but significant findings are also cited.This review focuses on development of lipopeptide vaccine systems including application of , bacterial lipopeptides, glycolipids and the lipid core peptide and their routes of administration. The use of liposomes as a delivery system that incorporates lipopeptides is discussed. The review also includes a brief description of immune system in relation to vaccinology and discussion on vaccine delivery routes.Lipids and their conjugates are an ideal frontrunner in the development of safe and efficient vaccines for different immunisation routes.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Keyword: immunity

Glucose enhances tilapia against Edwardsiella tarda infection through metabolome reprogramming.

We have recently reported that the survival of tilapia, Oreochromis niloticus, during Edwardsiella tarda infection is tightly associated with their metabolome, where the survived O.\xa0niloticus has distinct metabolomic profile to dying O.\xa0niloticus. Glucose is the key metabolite to distinguish the survival- and dying-metabolome. More importantly, exogenous administration of glucose to the fish greatly enhances their survival for the infection, indicating the functional roles of glucose in metabolome repurposing, known as reprogramming metabolomics. However, the underlying information for the reprogramming is not yet available. Here, GC/MS based metabolomics is used to understand the mechanisms by which how exogenous glucose elevates O.\xa0niloticus, anti-infectious ability to E.\xa0tarda. Results showed that exogenous glucose promotes stearic and biosynthesis but attenuates TCA cycle to potentiate O.\xa0niloticus against bacterial infection, which is confirmed by the fact that exogenous stearic increases immune protection in O.\xa0niloticus against E.\xa0tarda infection in a manner of Mx protein. These results indicate that exogenous glucose reprograms O.\xa0niloticus anti-infective metabolome that characterizes elevation of stearic and and attenuation of the TCA cycle. Therefore, our results proposed a novel mechanism that glucose promotes unsaturated fatty biosynthesis to cope with infection, thereby highlighting a potential way of enhancing fish in aquaculture.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: immunity

Identification of lipophilic ligands of Siglec5 and 14 that modulate innate immune responses.

Sialic -binding immunoglobulin-like lectins (Siglecs) are a family of cell-surface immune receptors that bind to sialic at terminal glycan residues. Siglecs also recognize non-sialic ligands, many of which remain to be characterized. Here, we found that Siglec5 and Siglec14 recognize lipid compounds produced by , a fungal genus containing several pathogenic species. Biochemical approaches revealed that the Siglec ligands are fungal alkanes and triacylglycerols, an unexpected finding that prompted us to search for endogenous lipid ligands of Siglecs. Siglec5 weakly recognized several endogenous lipids, but the mitochondrial lipid cardiolipin and the anti-inflammatory lipid 5--hydroxy stearic exhibited potent ligand activity on Siglec5. Further, the hydrophobic stretch in the Siglec5 N-terminus region was found to be required for efficient recognition of these lipids. Notably, this hydrophobic stretch was dispensable for recognition of sialic . Siglec5 inhibited cell activation upon ligand binding, and accordingly, the lipophilic ligands suppressed interleukin-8 (IL-8) production in Siglec5-expressing human monocytic cells. Siglec14 and Siglec5 have high sequence identity in the extracellular region, and Siglec14 also recognized the endogenous lipids. However, unlike Siglec5, Siglec14 transduces activating signals upon ligand recognition. Indeed, the endogenous lipids induced IL-8 production in Siglec14-expressing human monocytic cells. These results indicated that Siglec5 and Siglec14 can recognize lipophilic ligands that thereby modulate innate immune responses. To our knowledge, this is the first study reporting the binding of Siglecs to lipid ligands, expanding our understanding of the biological function and importance of Siglecs in the innate .Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: immunity

Mp1 Protein, a Novel Virulence Factor, Carries Two Arachidonic -Binding Domains To Suppress Inflammatory Responses in Hosts.

infection causes talaromycosis (previously known as penicilliosis), a very important opportunistic systematic mycosis in immunocompromised patients. Different virulence mechanisms in have been proposed and investigated. In the sera of patients with talaromycosis, Mp1 protein (Mp1p), a secretory galactomannoprotein antigen with two tandem ligand-binding domains (Mp1p-LBD1 and Mp1p-LBD2), was found to be abundant. Mp1p-LBD2 was reported to possess a hydrophobic cavity to bind copurified (PLM). It was hypothesized that capturing of lipids from human hosts by expressing a large quantity of Mp1p is a virulence mechanism of It was shown that expression of Mp1p enhanced the intracellular survival of by suppressing proinflammatory responses. Mechanistic study of Mp1p-LBD2 suggested that arachidonic (AA), a precursor of paracrine signaling molecules for regulation of inflammatory responses, is the major physiological target of Mp1p-LBD2. In this study, we use crystallographic and biochemical techniques to further demonstrate that Mp1p-LBD1, the previously unsolved first lipid binding domain of Mp1p, is also a strong AA-binding domain in Mp1p. These studies on Mp1p-LBD1 support the idea that the highly expressed Mp1p is an effective AA-capturing protein. Each Mp1p can bind up to 4 AA molecules. The crystal structure of Mp1p-LBD1-LBD2 has also been solved, showing that both LBDs are likely to function independently with a flexible linker between them. and potentially other pathogens highly expressing and secreting proteins similar to Mp1p can severely disturb host signaling cascades during proinflammatory responses by reducing the availabilities of important paracrine signaling molecules.Copyright © 2019 American Society for Microbiology.

Keyword: immunity

HDL inhibits saturated fatty mediated augmentation of innate immune responses in endothelial cells by a novel pathway.

Peripheral insulin resistance is associated with several metabolic abnormalities, including elevated serum fatty acids that contribute to vascular injury and atherogenesis. Our goals were to examine whether saturated fatty acids can modify innate immune responses to subclinical concentrations of lipopolysaccharide (LPS) in endothelial cells, and to explore the underlying pathway and determine whether it is modified by high density lipoprotein (HDL) and other factors commonly altered in insulin resistance.Physiologic concentrations of were added to human aortic endothelial cells with and without a variety of inhibitors or HDL and measures of cell inflammation and function assessed. significantly amplified human aortic endothelial cell inflammatory responses to LPS. Similar results were obtained from lipolysis products of triglyceride rich lipoproteins. Metabolism of to ceramide and subsequent activation of PKC-ζ, MAPK and ATF3 appeared critical in amplifying LPS induced inflammation. The amplified response to /LPS was decreased by HDL, dose dependently, and this inhibition was dependent on activation of PI3K/AKT and reduction in ATF3.These results indicate that endothelial cell innate immune responses are modified by metabolic abnormalities commonly present in insulin resistance and provide evidence for a novel mechanism by which HDL may reduce vascular inflammation.Copyright © 2016. Published by Elsevier B.V.

Keyword: immunity

Macrophage-Specific Hypoxia-Inducible Factor-1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis.

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia-inducible factors (HIFs) have recently been identified as important regulators of immunity and . In nonalcoholic steatohepatitis (NASH), HIF-1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF-1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF-1α levels in hepatic macrophages in methionine-choline-deficient (MCD) diet-fed mice and in macrophages of patients with NASH compared with controls. The HIF-1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin-1, LC3-II, and p62 in both mouse and human macrophages. LysM HIF fl/fl mice, which have HIF-1α levels stabilized in macrophages, showed higher steatosis and liver compared with HIF fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated fatty , (PA), both induces HIF-1α and impairs autophagic flux in macrophages. Using small interfering RNA-mediated knock-down and overexpression of HIF-1α in macrophages, we demonstrated that PA impairs autophagy via HIF-1α. We found that HIF-1α mediates NF-κB activation and MCP-1 production and that HIF-1α-mediated impairment of macrophage autophagy increases IL-1β production, contributing to MCD diet-induced NASH. Conclusion: impairs autophagy via HIF-1α activation in macrophages. HIF-1α and impaired autophagy are present in NASH in vivo in mouse macrophages and in human blood monocytes. We identified that HIF-1α activation and decreased autophagic flux stimulate in macrophages through upregulation of NF-κB activation. These results suggest that macrophage activation in NASH involves a complex interplay between HIF-1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet-induced NASH.© 2018 by the American Association for the Study of Liver Diseases.

Keyword: immunity

Lipidomic evidence that lowering the typical dietary palmitate to oleate ratio in humans decreases the leukocyte production of proinflammatory cytokines and muscle expression of redox-sensitive genes.

We recently reported that lowering the high, habitual (PA) intake in ovulating women improved insulin sensitivity and both inflammatory and oxidative stress. In vitro studies indicate that PA can activate both cell membrane toll-like receptor-4 and the intracellular nucleotide oligomerization domain-like receptor protein (NLRP3). To gain further insight into the relevance to human metabolic disease of dietary PA, we studied healthy, lean and obese adults enrolled in a randomized, crossover trial comparing 3-week, high-PA (HPA) and low-PA/high-oleic- (HOA) diets. After each diet, both hepatic and peripheral insulin sensitivities were measured, and we assessed cytokine concentrations in plasma and in supernatants derived from lipopolysaccharide-stimulated peripheral blood mononuclear cells (PBMCs) as well as proinflammatory gene expression in skeletal muscle. Insulin sensitivity was unaffected by diet. Plasma concentration of tumor necrosis factor-α was higher during the HPA diet. Lowering the habitually high PA intake by feeding the HOA diet resulted in lower secretion of interleukin (IL)-1β, IL-18, IL-10, and tumor necrosis factor-α by PBMCs, as well as lower relative mRNA expression of cJun and NLRP3 in muscle. Principal components analysis of 156 total variables coupled to analysis of covariance indicated that the mechanistic pathway for the differential dietary effects on PBMCs involved changes in the PA/OA ratio of tissue lipids. Our results indicate that lowering the dietary and tissue lipid PA/OA ratio resulted in lower leukocyte production of proinflammatory cytokines and muscle expression of redox-sensitive genes, but the relevance to diabetes risk is uncertain.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: immunity

Investment of both essential fatty and amino acids to varies depending on reproductive stage.

Trade-offs among the key life-history traits of reproduction and have been widely documented. However, the currency in use is not well-understood. We investigated how reproducing female side-blotched lizards, Uta stansburiana, allocate lipids versus proteins when given an immune challenge. We tested whether lizards would invest more in reproduction or depending on reproductive stage. Females were given stable isotopes ( N-leucine and C-1-), maintained on a regular diet and given either a cutaneous biopsy or a sham biopsy (control). Stable isotopes were monitored and analyzed in feces and uric , skin biopsies, eggs, and toe clips. We found that lizards deposited both proteins and lipids into their healing wounds (immune-challenged), skin (control), and eggs (all) and that catabolism of proteins exceeded incorporation into tissue during wound-healing. Specifically, we found that healed biopsies of wounded animals had more leucine and than the nonregrown skin biopsies taken from unwounded control animals. Earlier in reproduction, lizards invested relatively more labeled proteins into healing their wound tissue, but not into unwounded skin of control animals. Thus, reproduction is sometimes favored over self-maintenance, but only in later reproductive stages. Finally, we documented positive relationships among the amount of deposited in the eggs, the amount of food eaten, and the amount of excreted, suggesting higher turnover rates of lipids in lizards investing highly in their eggs.© 2019 Wiley Periodicals, Inc.

Keyword: immunity

Complement Component C3 Is Highly Expressed in Human Pancreatic Islets and Prevents β Cell Death via ATG16L1 Interaction and Autophagy Regulation.

We show here that human pancreatic islets highly express C3, which is both secreted and present in the cytosol. Within isolated human islets, C3 expression correlates with type 2 diabetes (T2D) donor status, HbA1c, and inflammation. Islet C3 expression is also upregulated in several rodent diabetes models. C3 interacts with ATG16L1, which is essential for autophagy. Autophagy relieves cellular stresses faced by β cells during T2D and maintains cellular homeostasis. C3 knockout in clonal β cells impaired autophagy and led to increased apoptosis after exposure of cells to and IAPP. In the absence of C3, autophagosomes do not undergo fusion with lysosomes. Thus, C3 may be upregulated in islets during T2D as a cytoprotective factor against β cell dysfunction caused by impaired autophagy. Therefore, we revealed a previously undescribed intracellular function for C3, connecting the complement system directly to autophagy, with a broad\xa0potential importance in other diseases and cell types.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: immunity

From and Vaccines to Mammalian Regeneration.

Our current understanding of major histocompatibility complex (MHC)-mediated antigen presentation in self and nonself immune recognition was derived from immunological studies of autoimmunity and virus-host interactions, respectively. The trimolecular complex of the MHC molecule, antigen, and T-cell receptor accounts for the phenomena of immunodominance and MHC degeneracy in both types of responses and constrains vaccine development. Out of such considerations, we developed a simple peptide vaccine construct that obviates immunodominance, resulting in a broadly protective T-cell response in the absence of antibody. In the course of autoimmunity studies, we identified the MRL mouse strain as a mammalian model of amphibian-like regeneration. A significant level of DNA damage in the cells from this mouse pointed to the role of the cell cycle checkpoint gene CDKN1a, or p21(cip1/waf1). The MRL mouse has highly reduced levels of this molecule, and a genetic knockout of this single gene in otherwise nonregenerating strains led to an MRL-type regenerative response, indicating that the ability to regenerate has not been lost during evolution.© The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Keyword: immunity

Transcriptomic and metabolomic analyses reveal that bacteria promote plant defense during infection of soybean cyst nematode in soybean.

Soybean cyst nematode (SCN) is the most devastating pathogen of soybean. Our previous study showed that the plant growth-promoting rhizobacterium Bacillus simplex strain Sneb545 promotes soybean resistance to SCN. Here, we conducted a combined metabolomic and transcriptomic analysis to gain information regarding the biological mechanism of defence enhancement against SCN in Sneb545-treated soybean. To this end, we compared the transcriptome and metabolome of Sneb545-treated and non-treated soybeans under SCN infection.Transcriptomic analysis showed that 6792 gene transcripts were common in Sneb545-treated and non-treated soybeans. However, Sneb545-treated soybeans showed a higher concentration of various nematicidal metabolites, including 4-vinylphenol, methionine, piperine, and , than non-treated soybeans under SCN infection.Overall, our results validated and expanded the existing models regarding the co-regulation of gene expression and metabolites in plants, indicating the advantage of integrated system-oriented analysis.

Keyword: immunity

Role of Sphingolipids in Infant Gut Health and Immunity.

Sphingomyelin (SM), glycosphingolipids, and gangliosides are important polar lipids in the milk fat globule membrane but are not found in standard milk replacement formulas. Because digestion and absorption of SM and glycosphingolipids generate the bioactive metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P), and because intact gangliosides may have beneficial effects in the gut, this may be important for gut integrity and immune maturation in the neonate. The brush border enzymes that hydrolyze milk SM, alkaline sphingomyelinase (nucleotide phosphodiesterase pyrophosphatase 7), and neutral ceramidase are expressed at birth in both term and preterm infants. Released sphingosine is absorbed, phosphorylated to S1P, and converted to via S1P-lyase in the gut mucosa. Hypothetically, S1P also may be released from absorptive cells and exert important paracrine actions favoring epithelial integrity and renewal, as well as\xa0immune function, including secretory IgA production and migration of T lymphocyte subpopulations. Gluco-, galacto-, and lactosylceramide are hydrolyzed to ceramide by lactase-phlorizin hydrolase, which also hydrolyzes lactose. Gangliosides may adhere to the brush border and is internalized, modified, and possibly transported into blood, and may exert protective functions by their interactions with bacteria, bacterial toxins, and the brush border.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: immunity

Sebum lipids influence macrophage polarization and activation.

As lipids are known to regulate macrophage functions, it is reasonable to suppose that a sebocyte-macrophage axis mediated by sebum lipids may exist.To investigate if sebocytes could contribute to the differentiation, polarization and function of macrophages with their secreted lipids.Oil Red O lipid staining and Raman spectroscopy were used to assess the dermal lipid content and penetration. Immunohistochemistry was used to analyse the macrophage subsets. Human peripheral blood monocytes were differentiated in the presence of either supernatant from human SZ95 sebocytes or major sebum lipid components and activated with Propionibacterium acnes. Macrophage surface markers and their capacity to uptake fluorescein isothiocyanate-conjugated P.\xa0acnes were detected by fluorescence-activated cell sorting measurements. Cytokine protein levels were evaluated by enzyme-linked immunosorbent assay and Western blot analysis.Sebaceous gland-rich skin had an increased dermal lipid content vs. sebaceous gland-poor skin to which all the tested sebum component lipids could contribute by penetrating the dermoepidermal barrier. Of the lipids, oleic and linoleic promoted monocyte differentiation into alternatively activated macrophages. Moreover, linoleic also had an anti-inflammatory effect in P.\xa0acnes-activated macrophages, inhibiting the secretion of interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α. Squalene, , stearic and oleic augmented the secretion of IL-1β, even in the absence of P. acnes, whereas oleic had a selective effect of inducing IL-1β but downregulating IL-6 and TNF-α secretion.Our results suggest a role for sebaceous glands in modulating innate immune responses via their secreted lipids that are of possible pathological and therapeutic relevance.© 2017 British Association of Dermatologists.

Keyword: immunity

Isoliquiritigenin Attenuates Adipose Tissue Inflammation in vitro and Adipose Tissue Fibrosis through Inhibition of Innate Immune Responses in Mice.

Isoliquiritigenin (ILG) is a flavonoid derived from Glycyrrhiza uralensis and potently suppresses NLRP3 inflammasome activation resulting in the improvement of diet-induced adipose tissue inflammation. However, whether ILG affects other pathways besides the inflammasome in adipose tissue inflammation is unknown. We here show that ILG suppresses adipose tissue inflammation by affecting the paracrine loop containing saturated fatty acids and TNF-α by using a co-culture composed of adipocytes and macrophages. ILG suppressed inflammatory changes induced by the co-culture through inhibition of NF-κB activation. This effect was independent of either inhibition of inflammasome activation or activation of peroxisome proliferator-activated receptor-γ. Moreover, ILG suppressed TNF-α-induced activation of adipocytes, coincident with inhibition of IκBα phosphorylation. Additionally, TNF-α-mediated inhibition of Akt phosphorylation under insulin signaling was alleviated by ILG in adipocytes. ILG suppressed -induced activation of macrophages, with decreasing the level of phosphorylated Jnk expression. Intriguingly, ILG improved high fat diet-induced fibrosis in adipose tissue in vivo. Finally, ILG inhibited TLR4- or Mincle-stimulated expression of fibrosis-related genes in stromal vascular fraction from obese adipose tissue and macrophages in vitro. Thus, ILG can suppress adipose tissue inflammation by both inflammasome-dependent and -independent manners and attenuate adipose tissue fibrosis by targeting innate immune sensors.

Keyword: immunity

Investigating Cellular Quiescence of T Lymphocytes and Antigen-Induced Exit from Quiescence.

Naïve T cells are in a quiescent state under homeostasis but respond to antigen stimulation by exiting from quiescence and entering the cell cycle. Appropriate regulation of quiescence is crucial for maintaining T cell homeostasis at steady state and initiating proper T cell responses to antigen stimulation. Emerging evidence indicates that signaling by mechanistic target of rapamycin (mTOR) plays a central role in the control of T cell quiescence and antigen-induced exit from quiescence through coordinating immune signals, cellular metabolic programs, and cell cycle machinery. The mTOR-dependent regulation of quiescence has also been implicated in the differentiation and function of memory T cells. In this chapter, we describe techniques to assess quiescent state of naïve T cells under steady state and exit from quiescence upon TCR stimulation.

Keyword: immunity

Elevated biosynthesis of is required for zebrafish against Edwardsiella tarda infection.

Mechanisms by which vaccines enhance to combat bacterial pathogens are not fully understood. Recently, we have found that live Edwardsiella tarda vaccine enhances ability against the bacterial challenge by metabolic modulation in zebrafish. Here we first explored the metabolic modulation promoted by inactivated E. tarda to eliminate the pathogen. Inactivated E. tarda vaccine modulated a similar metabolome to combat with the pathogen in zebrafish as live E. tarda vaccine did. Specifically, both vaccines promoted biosynthesis of unsaturated fatty acids and the TCA cycle. However, due to relatively higher activated TCA cycle in inactivated vaccine than live vaccine, live vaccine promoted higher abundance of palmitate than inactivated vaccine. Consistently, the protection against E. tarda challenge was palmitate dose-dependent. Live vaccine activated higher expression of IL-1β, IL-8,Cox-2 genes and lower expression of IL-15, IL-21 genes than inactivated vaccine, which is similar to the results stimulated by high and low doses of palmitate, respectively. These findings indicate live and inactivated E. tarda vaccines stimulate differential abundances of palmitate that contribute to differential innate immunities against bacterial infection. Thus, metabolic environment contributes to immune response.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: immunity

Effects of elevated growth temperature and heat shock on the lipid composition of the inner and outer membranes of Yersinia pseudotuberculosis.

Differences in the distribution of individual phospholipids between the inner (IM) and outer membranes (OM) of gram-negative bacteria have been detected in mesophilic Escherichia, Erwinia and Salmonella species but have never been investigated in the psychrotrophic Yersinia genus. Therefore, the influence of an elevated growth temperature and heat shock on the phospholipid and fatty (FA) compositions of the fractionated Yersinia pseudotuberculosis envelope was investigated. The shift of the growth temperature from 8\xa0°C to 37\xa0°C to mimic the switch from saprophytic to parasitic growth of this bacteria and the exposure of the cells to heat shock, which was induced by a sharp increase in the temperature from 8\xa0°C to 45\xa0°C, increased the lysophosphatidylethanolamine content from zero and 1% to 6% and 10% in the IM and OM, respectively. These changes were accompanied by a decrease in the phosphatidylethanolamine (PE) content and a drastic increase (up to 3-fold higher) in the phosphatidylglycerol (PG) level in the OM of the bacteria, which increases the net negative charge of the cell envelope. The levels of the predominant saturated (16:0) and cyclopropane FAs were approximately 1.5- and 7.5-fold higher, respectively, but the content of the predominant unsaturated palmitoleic (16:1n-7) and cis-vaccenic (18:1n-7) FAs was approximately 10-30-fold lower in both membranes that were isolated from the cells grown at elevated temperatures. Due to these changes, reflecting the process of "homeoviscous adaptation", the ratio between the unsaturated and saturated FAs decreased but remained higher in the IM than that in the OM. Simultaneously, no significant changes were observed in the FA composition of cells subjected to heat shock, demonstrating a difference between the responses of the heat-shocked and heat-adapted Y.\xa0pseudotuberculosis. The unique ability of Y.\xa0pseudotuberculosis to reciprocally regulate the ratio of anionic PG and net neutral PE and therefore adjust the negative charge of the OM may be a common strategy used by pathogenic bacteria to promote the barrier function of the OM.Copyright © 2016. Published by Elsevier B.V.

Keyword: immunity

Fatty status in infancy is associated with the risk of type 1 diabetes-associated autoimmunity.

We investigated the association of early serum fatty composition with the risk of type 1 diabetes-associated autoimmunity. Our hypothesis was that fatty status during infancy is related to type 1 diabetes-associated autoimmunity and that long-chain n-3 fatty acids, in particular, are associated with decreased risk.We performed a nested case-control analysis within the Finnish Type 1 Diabetes Prediction and Prevention Study birth cohort, carrying HLA-conferred susceptibility to type 1 diabetes (n\xa0=\xa07782). Serum total fatty composition was analysed by gas chromatography in 240 infants with islet autoimmunity and 480 control infants at the age of 3 and 6\xa0months. Islet autoimmunity was defined as repeated positivity for islet cell autoantibodies in combination with at least one of three selected autoantibodies. In addition, a subset of 43 infants with primary insulin autoimmunity (i.e. those with insulin autoantibodies as the first autoantibody with no concomitant other autoantibodies) and a control group (n\xa0=\xa086) were analysed. A third endpoint was primary GAD autoimmunity defined as GAD autoantibody appearing as the first antibody without other concomitant autoantibodies (22 infants with GAD autoimmunity; 42 infants in control group). Conditional logistic regression was applied, considering multiple comparisons by false discovery rate <0.05.Serum fatty composition differed between breastfed and non-breastfed infants, reflecting differences in the fatty composition of the milk. Fatty acids were associated with islet autoimmunity (higher serum pentadecanoic, , palmitoleic and docosahexaenoic acids decreased risk; higher arachidonic:docosahexaenoic and n-6:n-3 ratios increased risk). Furthermore, fatty acids were associated with primary insulin autoimmunity, these associations being stronger (higher palmitoleic , cis-vaccenic, arachidonic, docosapentaenoic and docosahexaenoic acids decreased risk; higher α-linoleic and arachidonic:docosahexaenoic and n-6:n-3 ratios increased risk). Moreover, the quantity of breast milk consumed per day was inversely associated with primary insulin autoimmunity, while the quantity of cow\'s milk consumed per day was directly associated.Fatty status may play a role in the development of type 1 diabetes-associated autoimmunity. Fish-derived fatty acids may be protective, particularly during infancy. Furthermore, fatty acids consumed during breastfeeding may provide protection against type 1 diabetes-associated autoimmunity. Further studies are warranted to clarify the independent role of fatty acids in the development of type 1 diabetes.

Keyword: immunity

Direct interaction, instrumental for signaling processes, between LacCer and Lyn in the lipid rafts of neutrophil-like cells.

Lactosylceramide [LacCer; β-Gal-(1-4)-β-Glc-(1-1)-Cer] has been shown to contain very long fatty acids that specifically modulate neutrophil properties. The interactions between LacCer and proteins and their role in cell signaling processes were assessed by synthesizing two molecular species of azide-photoactivable tritium-labeled LacCer having acyl chains of different lengths. The lengths of the two acyl chains corresponded to those of a short/medium and very long fatty , comparable to the lengths of stearic and lignoceric acids, respectively. These derivatives, designated C18-[(3)H]LacCer-(N3) and C24-[(3)H]LacCer-(N3), were incorporated into the lipid rafts of plasma membranes of neutrophilic differentiated HL-60 (D-HL-60) cells. C24-[(3)H]LacCer-(N3), but not C18-[(3)H]LacCer-(N3), induced the phosphorylation of Lyn and promoted phagocytosis. Incorporation of C24-[(3)H]LacCer-(N3) into plasma membranes, followed by illumination, resulted in the formation of several tritium-labeled LacCer-protein complexes, including the LacCer-Lyn complex, into plasma membrane lipid rafts. Administration of C18-[(3)H]LacCer-(N3) to cells, however, did not result in the formation of the LacCer-Lyn complex. These results suggest that LacCer derivatives mimic the biological properties of natural LacCer species and can be utilized as tools to study LacCer-protein interactions, and confirm a specific direct interaction between LacCer species containing very long fatty acids, and Lyn protein, associated with the cytoplasmic layer via myristic/ chains.Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: immunity

Blockade of myeloid differentiation protein 2 prevents obesity-induced inflammation and nephropathy.

Obesity is a major and independent risk factor of kidney diseases. The pathogenic mechanisms of obesity-associated renal injury are recognized to at least involve a lipid-rich and pro-inflammatory state of the renal tissues, but specific mechanisms establishing causal relation remain unknown. Saturated fatty acids are elevated in obesity, and known to induce chronic inflammation in kidneys. Myeloid differentiation protein 2 (MD2) is an important protein in lipopolysaccharide-induced innate response and inflammation. We suggested that obesity-associated renal injury is regulated by MD2 thereby driving an inflammatory renal injury. The used three mouse models for in vivo study: MD2 knockout mice (KO) maintained on high fat diet (HFD), wild-type mice on HFD plus L6H21, a specific MD2 inhibitor and KO mice given (PA) by IV injection. The in vitro studies were carried out in cultured renal tubular epithelial cells, mouse mesangial cells and primary macrophages, respectively. The HFD mice presented with increased hyperlipidemia, serum creatinine and proteinuria. Renal tissue from HFD mice had increased fibrosis, inflammatory cytokines, macrophage infiltration, and activation of NF-κB and MAPKs. This HFD-induced renal injury profile was not observed in KO mice or L6H21-treated mice. Mice given PA mimmicked the HFD-induced renal injury profiles, which were prevented by MD2 knockout. The in vitro data further confirmed MD2 mediates PA-induced inflammation. MD2 is causally related with obesity-associated renal inflammatory injury. We believe that MD2 is an attractive target for future therapeutic strategies in obesity-associated kidney diseases.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: immunity

Surface engineering of Solid Lipid Nanoparticle assemblies by methyl α-d-mannopyranoside for the active targeting to macrophages in anti-tuberculosis inhalation therapy.

This study describes the development of new mannosylated Solid Lipid Nanoparticle assemblies (SLNas) delivering rifampicin for an inhaled treatment of tuberculosis. SLNas were surface engineered with mannose residues to recognize mannose receptors located on infected alveolar macrophages and facilitate cell internalization. Two sets of SLNas were produced by the melt emulsifying technique using biocompatible lipid components, i.e. cholesteryl myristate combined with (PA set) or tripalmitin (TP set), in the presence of the targeting moiety, methyl α-d-mannopyranoside. Mannosylated SLNas were examined for their physical properties, drug payloads and release, as well as respirability in terms of emitted dose and respirable fraction determined by Next Generation Impactor. The most appropriate formulations were assessed for mannosylation using FTIR, XPS, SEM coupled with EDX analysis, and wettability assay, in comparison with the respective non-functionalized SLNas. Besides, cytotoxicity and cell internalization ability were established on J774 murine macrophage cell line. Mannosylated SLNas exhibited physical properties suitable for alveolar macrophage passive targeting, adequate rifampicin payloads (10-15%), and feasible drug maintenance within SLNas along the respiratory tract before macrophage internalization. Despite respirability impaired by powder cohesiveness, surface mannosylation provided quicker macrophage phagocytosis, giving evidence of an active targeting promotion.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: immunity

Sex differences in the phagocytic and migratory activity of microglia and their impairment by .

Sex differences in the incidence, clinical manifestation, disease course, and prognosis of neurological diseases, such as autism spectrum disorders or Alzheimer\'s disease, have been reported. Obesity has been postulated as a risk factor for cognitive decline and Alzheimer\'s disease and, during pregnancy, increases the risk of autism spectrum disorders in the offspring. Obesity is associated with increased serum and brain levels of free fatty acids, such as , which activate microglial cells triggering a potent inflammatory cascade. In this study, we have determined the effect of in the inflammatory profile, motility, and phagocytosis of primary male and female microglia, both in basal conditions and in the presence of a pro-inflammatory stimulus (interferon-γ). Male microglia in vitro showed higher migration than female microglia under basal and stimulated conditions. In contrast, female microglia had higher basal and stimulated phagocytic activity than male microglia. did not affect basal migration or phagocytosis, but abolished the migration and phagocytic activity of male and female microglia in response to interferon-γ. These findings extend previous observations of sex differences in microglia and suggest that impairs the protective responses of these cells.© 2017 Wiley Periodicals, Inc.

Keyword: immunity

Increased palmitate intake: higher acylcarnitine concentrations without impaired progression of β-oxidation.

(PA) is associated with higher blood concentrations of medium-chain acylcarnitines (MCACs), and we hypothesized that PA may inhibit progression of FA β-oxidation. Using a cross-over design, 17 adults were fed high PA (HPA) and low PA/high oleic (HOA) diets, each for 3 weeks. The [1-(13)C]PA and [13-(13)C]PA tracers were administered with food in random order with each diet, and we assessed PA oxidation (PA OX) and serum AC concentration to determine whether a higher PA intake promoted incomplete PA OX. Dietary PA was completely oxidized during the HOA diet, but only about 40% was oxidized during the HPA diet. The [13-(13)C]PA/[1-(13)C]PA ratio of PA OX had an approximate value of 1.0 for either diet, but the ratio of the serum concentrations of MCACs to long-chain ACs (LCACs) was significantly higher during the HPA diet. Thus, direct measurement of PA OX did not confirm that the HPA diet caused incomplete PA OX, despite the modest, but statistically significant, increase in the ratio of MCACs to LCACs in blood.Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: immunity

The N-Acylethanolamine Amidase Inhibitor ARN077 Suppresses and Pruritus in a Mouse Model of Allergic\xa0Dermatitis.

N-acylethanolamine amidase (NAAA), a cysteine hydrolase highly expressed in macrophages and B lymphocytes, catalyzes the degradation of palmitoylethanolamide. Palmitoylethanolamide is an agonist of PPAR-α and an important regulator of pain and innate immunity. In this study, we investigated the properties of the NAAA inhibitor, ARN077, in a mouse model of allergic contact dermatitis. Acute topical applications of ARN077 attenuated key signs of DNFB-induced dermatitis in a dose-dependent manner. Moreover, ARN077 increased tissue palmitoylethanolamide content and normalized circulating levels of cytokines and immunoglobulin E. No such effect was seen in PPAR-α-deficient mice. Moreover, mice lacking NAAA failed to develop edema or scratching behavior after challenge with DNFB, confirming that this enzyme plays an important role in dermatitis. Consistent with this conclusion, subchronic applications of ARN077 suppressed DNFB-induced when administered either before or after the DNFB challenge. The effects of subchronic ARN077 were dose dependent and comparable in size to those produced by the steroids clobetasol and dexamethasone. Unlike the latter, however, ARN077 did not cause skin atrophy. The results identify NAAA as a promising target for the development of effective and safe treatments for atopic dermatitis and other inflammatory disorders of the skin.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: immunity

Molecular characterization, functional expression, tissue localization and protective potential of a Taenia solium fatty -binding protein.

The fatty -binding proteins (FABPs) comprise a family of proteins that are widely expressed in animal cells and perform a variety of vital functions. Here, we report the identification, characterization, recombinant expression, tissue localization and protective potential of a Taenia solium FABP (TsFABP1). The TsFABP1 primary structure showed all the conserved residues characteristic of the subfamily iv of the intracellular Lipid-Binding Proteins (iLBPs), including those involved in the binding stabilization of the fatty molecule. Through a competitive binding assay we found that TsFABP1 is able to bind at least six different fatty acids with preference toward and stearic , suggesting that TsFABP1 is a member of the iLBP subfamily iv. Immunolocalization assays carried out on larval and adult tissues of four species of taeniids using anti-TsFABP1 hyperimmune sera produced in mice and rabbit, showed intense labeling in the tegument of the spiral canal and in subtegumental cytons of the larvae. These findings suggest that the spiral canal might be a major place for FA uptake in the developing scolex. In contrast, only subtegumental cytons in the adult worms stained positive. We propose that TsFABP1 is involved in the mechanism to mobilize fatty acids between compartments in the extensive syncytial tissue of taeniids. Protection assays carried out in a murine model of cysticercosis showed that subcutaneous immunization with TsFABP1 resulted in about 45% reduction of parasite load against an intraperitoneal challenge with Taenia crassiceps cysts. This reduction in parasite load correlated with the level of cellular and humoral immune responses against TsFABP1, as determined in spleen lymphocyte proliferation and ELISA testing.Copyright © 2012 Elsevier B.V. All rights reserved.

Keyword: immunotherapy

Inducing protective antibodies against ring-infected erythrocyte surface peptide antigen of Plasmodium falciparum using immunostimulating complex (ISCOMs) delivery.

In the present study, synthetic peptides (EENVEHDA)2 [(oc)2] and (DDEHVEEPTVA)2 [(un)2] of ring-infected erythrocyte surface antigen (RESA) of Plasmodlium filciparum were linked with and entrapped in immunostimulating complexes (ISCOMs). The immunogenicity of the peptide(s) and mixture of peptides were studied in mice with different genetic background. Peptide(s) entrapped in ISCOMs using a low-dose immunization strategy generated high-titer as well as high-affinity antibodies. Interestingly, no genetic restriction of the immune response was observed in any of the strains studied. The IgG subclass pattern with the peptide(s) showed predominately IgG2a/2b isotypes, while with the mixed peptide formulation, (un)2-specific IgG isotype pattern showed induction of both IgG1 and IgG2a/2b isotypes. These cytophilic antibodies inhibited the ring as well as schizont stage and total parasite growth during in vitro merozoite reinvasion inhibition study. In the mixed peptide preparation, the same pattern of immune response was achieved as that of individual peptide(s) using ISCOMs delivery. Therefore, the entrapment of otherwise poorly immunogenic synthetic peptides in ISCOMs resulted in increased immunogenicity followed by strong secondary response and can be adopted for developing subunit immunogen formulation against malarial parasite.

Keyword: immunotherapy

A palmitoyl-tailed sequential oligopeptide carrier for engineering immunogenic conjugates.

The main guideline in designing effective immunogens as vaccine candidates capable of eliciting potent and specific immune responses is to combine B/T cell epitopes and adjuvants as immunostimulators on the same carrier that links the major histocompatibility complex with T cell receptors. Aiming at contributing to the development of carriers for human usage a helicoid type sequential oligopeptide carrier, SOC(n)-II, formed by the repeating tetrapeptide unit (Aib-Lys-Aib-Gly)(n), n=2-7, elongated from the amino-terminus by the palmitoyl group, known for its adjuvanticity, is now presented. The main B cell epitope, PPGMRPP, of the Sm autoantigen against which the majority of antibodies in patients with systemic lupus erythematosus is directed, was coupled to the Lys-N(epsilon)H(2) groups of the carrier in four copies and the resulting conjugate Palm-SOC(4)-II-Sm(4) was subjected to animal immunizations without utilizing any adjuvant. The induced immune response was comparable with that produced when Ac-SOC(4)-II-Sm(4) was administered in animals following the conventional immunization protocol of complete/incomplete Freund\'s adjuvant. High titers of anti-Palm-SOC(4)-II-Sm(4) antibodies were generated, which recognize the priming immunogenic conjugate, as well as reconstituted Sm mimics but not the carrier alone. It is concluded that Palm-SOC(n)-II carrier is a valuable tool for engineering immunogens eliciting enhanced and specific humoral immune responses.

Keyword: immunotherapy

Depletion of tumor-associated macrophages enhances the anti-tumor immunity induced by a Toll-like receptor agonist-conjugated peptide.

It has been reported that lipopeptides can be used to elicit cytotoxic T lymphocyte (CTL) responses against viral diseases and cancer. In our previous study, we determined that mono-palmitoylated peptides can enhance anti-tumor responses in the absence of adjuvant activity. To investigate whether di-palmitoylated peptides with TLR2 agonist activity are able to induce anti-tumor immunity, we synthesized a di--conjugated long peptide that contains a murine CTL epitope of HPV E749-57 (Pam2IDG). Pam2IDG stimulated the maturation of bone marrow-derived dendritic cells (BMDCs) through TLR2/6. After immunization, Pam2IDG induced higher levels of T cell responses than those obtained with its non-lipidated counterpart (IDG). In the prophylactic model, Pam2IDG immunization completely inhibited tumor growth, whereas IDG immunization was unable to inhibit tumor growth. However, Pam2IDG immunization could not effectively inhibit the growth of established tumors. Therefore, we further investigated whether the depletion of immunosuppressive factors could improve the therapeutic effects of Pam2IDG. Our data indicate that treatment with Pam2IDG combined with clodronate/liposome delays tumor growth and increases the survival rate. We also observed that the therapeutic effects of Pam2IDG are improved by diminishing the function of tumor-associate macrophages (TAMs) and through the use of an IL10 receptor blocking antibody or a Cyclooxygenase 2 (Cox-2) inhibitor. In conclusion, the depletion of TAMs may enhance the anti-tumor immunity of a TLR2 agonist-conjugated peptide.

Keyword: immunotherapy

Lipopeptide epitopes extended by an Nepsilon-palmitoyl-lysine moiety increase uptake and maturation of dendritic cells through a Toll-like receptor-2 pathway and trigger a Th1-dependent protective immunity.

Lipopeptides, a form of peptide immunogens, are currently under intense investigation as human vaccines for many infectious pathogens and cancers. However, the cellular and molecular mechanisms of lipopeptide immunogenicity are only partially understood. We have investigated the influence of the lipid content on the immunogenicity of lipopeptides using the herpes simplex virus type 1 (HSV-1) gD(1-23) peptide as a model antigen. Totally synthetic lipopeptides were constructed by covalent attachment to the peptide backbone of either Nepsilon-palmitoyl-lysine (palmitoyl-lipidated peptide, palmitoyl-LP) or cholesterol-lysine (cholesterol-lipidated peptide, cholesterol-LP). Immunization of mice with the palmitoyl-LP, but not with its cholesterol-LP analog, induced a strong T cell-dependent protective immunity against lethal HSV-1 infection. Analysis of cytokine profiles and IgG2a/IgG1 ratios revealed that a dominant Th1-type immune response was stimulated by the palmitoyl-LP, as opposed to a Th2 response generated by its cholesterol-LP analog. The palmitoyl-LP was efficiently taken up in vitro by immature dendritic cells (DC) in a time- and dose-dependent manner, and induced phenotypic maturation and production of pro-inflammatory cytokines by DC. Finally, DC stimulated with the palmitoyl-LP induced antigen-specific T cell responses through the Toll-like receptor-2 pathway. These findings have important implications for the development of effective lipopeptide immunization strategies against infectious pathogens.

Keyword: immunotherapy

Desensitization, phosphorylation and palmitoylation of the human dopamine D1 receptor.

The regulation and post-translational modifications of the human dopamine D1 receptor were studied in the baculovirus-eukaryotic cell expression system. Baculovirus constructs containing either the DNA encoding the dopamine D1 receptor or a DNA encoding a c-myc epitope tagged dopamine D1 receptor (c-myc-dopamine D1 receptor) were used to infect Spodoptera frugiperda (Sf9) insect cells. Expressed dopamine D1 and c-myc-dopamine D1 receptors bound agonists and antagonists with affinities and a rank order of potency characteristic of a classical dopamine D1 receptor pharmacological profile. In membrane preparations from cells expressing c-myc-dopamine D1 receptor, the photoaffinity label [125I](3-methyl-2-[4\'-azidophenyl]-2,3,5-tetrahydro-2H-3-benzazepine) ([125I]MAB) bound specifically upon photolysis. A major broad band of approximately 48 kDa was detected. This species was identified in immunoblots by the monoclonal antibody raised against the c-myc epitope of c-myc-dopamine D1 receptor was isolated by immunoprecipitation from whole cells and was shown to be post-translationally modified by phosphorylation and palmitoylation. Exposure of cells expressing c-myc-dopamine D1 receptor to dopamine for 15 min resulted in a reduction in the maximal dopamine stimulated adenylyl cyclase activity, which was accompanied by an increased phosphorylation of the receptor and a rapid redistribution of surface c-myc-dopamine D1 receptor as detected by in situ immunofluorescence. Dopamine exposure also resulted in an increased level of incorporation of [3H] into the receptor. Thus, we provide the first evidence that the human dopamine D1 receptor undergoes agonist-dependent desensitization, phosphorylation and palmitoylation.

Keyword: immunotherapy

Development of a lipopeptide-based therapeutic vaccine to treat chronic HBV infection. I. Induction of a primary cytotoxic T lymphocyte response in humans.

Our goal is to use peptide epitopes that are recognized by cytotoxic T lymphocytes (CTL) as immunogens for the development of prophylactic and therapeutic vaccines with chronic hepatitis B virus (HBV) infection being our first therapeutic target. Because most CTL peptide epitopes are poor immunogens, we specifically modified them by covalently attaching two additional components: a T helper peptide epitope and two lipid molecules. Using the murine influenza virus CTL epitope NP 147-155 as a model system, we found this construct to be highly immunogenic, and a single injection resulted in memory CTL induction that persisted for > 1 yr. Based on the animal studies, a vaccine was designed and tested for both safety and its ability to induce a primary CTL response in normal subjects. The three vaccine components included HBV core antigen peptide 18-27 as the CTL epitope, tetanus toxoid peptide 830-843 as the T helper peptide, and two molecules as the lipids. A dose escalation trial (5, 50, and 500 micrograms) carried out in 26 normal subjects showed that the vaccine was safe and able to induce a primary HBV-specific CTL response. A dose-response curve was observed and five out of five subjects responded to the 500-micrograms dose.

Keyword: immunotherapy

Designer cancer vaccines made easy: protein transfer of immunostimulatory molecules for use in therapeutic tumor vaccines.

Advances in the understanding of the immune response to tumors has led to the development of new strategies to design therapeutic vaccines. One of these strategies is the development of protein transfer of immunostimulatory molecules onto the surface of tumor cells, thereby directing the immune response to the tumor antigens carried by the modified tumor cells. This strategy has been developed as an alternative to gene transfer, the more classical technique of introducing immunostimulatory molecules onto tumors. In this report we briefly review current strategies for and then focus on several approaches to protein transfer and their historical basis. Finally, the application of these protein transfer approaches to develop cancer vaccines are reviewed and discussed.

Keyword: immunotherapy

Pools of lipidated HTL-CTL constructs prime for multiple HBV and HCV CTL epitope responses.

Various peptide-based approaches to simultaneous induction of multiple cytotoxic T lymphocyte (CTL) responses were evaluated as part of ongoing efforts to develop immunotherapeutic vaccines for use in humans. To this end, HLA (human histocompatibility leukocyte antigen)-A2-restricted epitopes from several specific viral proteins were tested in an HLA-A2 transgenic mouse model system, which mimics human CTL responses to these viral proteins. Multiple CTL responses were elicited by immunization with either peptides emulsified in incomplete Freund\'s adjuvant (IFA), or lipidated peptides administered in phosphate buffered saline (PBS). In the case of lipidated peptides, induction of CTL responses was crucially dependent on the presence of helper T lymphocyte (HTL) epitopes, and most efficient in the case of lipidated covalently linked HTL-CTL epitope constructs. CTL could also be induced by immunization with lipidated HTL epitopes simply mixed with CTL epitopes and formulated in PBS. However, this approach was highly dependent on the particular lipidated HTL/CTL combination utilized, and was marginally effective for simultaneous priming of multiple CTL responses. By contrast, all HTL/CTL combinations were potent immunogens when delivered as lipidated, covalently linked molecules. This was the most effective of the approaches analysed in terms of multi-epitope priming, as demonstrated by the induction of simultaneous CTL responses to a pool of five different epitopes.

Keyword: immunotherapy

Protective immunity against ocular herpes infection and disease induced by highly immunogenic self-adjuvanting glycoprotein D lipopeptide vaccines.

An important phase in the development of an ocular herpes simplex virus type 1 (HSV-1) subunit vaccine is the identification of an efficient, safe, and adjuvant-free antigen delivery system capable of inducing and sustaining long-term memory T-cell protective immunity. This study was conducted to test the hypothesis that immunization with self-adjuvanting lipopeptide bearing HSV-1 glycoprotein D (gD) T-cell epitopes would elicit long-term HSV-specific T cells and decrease infection, disease, or both in a ocular herpes mouse model.Five immunodominant CD4(+) T-cell peptide epitopes (gD(1-29), gD(49-82), gD(146-179), gD(228-257), and gD(332-358)), recently identified from HSV-1 gD, were covalently linked to a moiety (lipopeptides) and delivered subcutaneously in adjuvant-free saline. The primary and memory T cells induced by these molecularly defined lipopeptides and their protective efficacy were assessed, in terms of virus replication in the eye, ocular disease, and survival.Three gD lipopeptides, that drive dendritic cell maturation in vitro, induced long-term, virus-specific, IFN-gamma-producing CD4(+) Th(1) responses, associated with a reduction in ocular herpes infection and disease. Immunization with a cocktail of these three highly immunogenic Th(1) lipopeptides increased survival, lowered the peak of ocular virus titer, and cleared the ocular disease.Vaccination with a mixture self-adjuvanting lipopeptides containing novel HSV-1 immunodominant gD T-cell epitopes protected mice from ocular herpes infection and disease. The strength of protective immunity induced by these lipopeptides together with their safety provide a molecularly defined vaccine formulation that could combat ocular herpes infection and disease in humans.

Keyword: immunotherapy

Novel lipopeptides of ESAT-6 induce strong protective immunity against Mycobacterium tuberculosis: Routes of immunization and TLR agonists critically impact vaccine\'s efficacy.

Mycobacterium tuberculosis (Mtb), the bacterial cause of tuberculosis, is a leading infectious agent worldwide. The development of a new vaccine against Mtb is essential to control global spread of tuberculosis, since the current vaccine BCG is not very effective and antibiotic resistance is a serious, burgeoning problem. ESAT-6 is a secreted protein of Mtb, which is absent in BCG but has been implicated in inducing protective immunity against Mtb. Peptide based subunit vaccines are attractive due to their safety and high specificity in eliciting immune responses, but small synthetic peptides are usually not very immunogenic. We have designed a novel subunit vaccine for Mtb by using simple lipid () modified derivatives of peptides from ESAT-6 protein corresponding to dominant human T cell epitopes and examined their ability to stimulate protective immunity against Mtb by intranasal and subcutaneous immunization in mice. We also investigated how individual TLR agonists as adjuvants (PolyI:C, MPL and GDQ) contribute to enhancing the induced immune responses and resulting protective efficacy of our vaccine. We observed that single C-terminal palmitoyl-lysine modified lipopeptides derived from ESAT-6 induce significant cellular immune responses on their own upon mucosal and subcutaneous immunizations. Intriguingly, a combination of immunogenic lipopeptides of ESAT-6 antigen exhibited local (pulmonary) and systemic immune responses along with efficient protective efficacy when administered intranasally or subcutaneously. Surprisingly, combination of ESAT-6 derived lipopeptides with a TLR-4 agonist (MPL) enhanced protection, whereas TLR-3 (Poly I:C) and TLR-7/8 agonists (gardiquimod, GDQ) led to reduced protection associated with specific local and systemic immune modulation. Our studies demonstrate the potential of ESAT-6 derived lipopeptides as a promising vaccine candidate against Mtb, and emphasize that selection of adjuvant is critical for the success of vaccines. These findings demonstrate the promise of synthetic lipopeptides as the basis of a subunit vaccine for TB.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: immunotherapy

[Establishment of an immunological labelling of pigs using synthetic peptides].

The objective of this study was to develop a suitable combination consisting of synthetic peptides, carrier protein and adjuvant for immunological labelling of pigs. Specific antibody titres were evaluated by ELISA technique. From 9 peptides 4 were excluded from following investigations showing cross reactivity or low immunogenic effects, sufficient anti-peptide titres were achieved by 5 peptides. Labelling control is possible after 7 days at the earliest and can be used for the whole fattening period after single immunization using an effective carrier-adjuvant-combination. Mixing of peptides in one labelling dose had no negative influence on titres against single peptides. Different combinations of 4 carriers (keyhole limpet haemocyanin (KLH), -3-cystein- (Pam3Cys), (Pam) and dextran) and 4 adjuvants (Montanide IMS 1313, Montanide ISA 25, Quil A and Diluvac forte) were tested. Optimal labelling could be seen by combination of 50 nmol peptide, KLH as carrier and Montanide IMS 1313 as adjuvant. Generally after booster injection titres were higher, however, a booster dose was not necessary using most effective adjuvants. A less immunogenic, but cost effective alternative for short time labelling (7 weeks) was a peptide-Pam3Cys-conjugate (75 nmol) combined with Quil A as an adjuvant (2 mg/ml). Immunological labelling of pigs is recommended as a good method for tracing back the origin of animals and meat products. It may be also used for vaccine labelling to prove vaccination of pigs.

Keyword: immunotherapy

Immune responses and protective effect in mice vaccinated orally with surface sporozoite protein of Eimeria falciformis in ISCOMs.

Immunostimulating complexes (ISCOMs) were built after treatment of a purified surface protein from Eimeria falciformis sporozoites with a derivation, leading to a high ratio (33-64%) of P27 incorporation in these cage-like structures. P27 kept its antigenicity after incorporation in ISCOMs, which induced, after iterative intubations by the oral route to groups of mice, a systemic IgG response, a local IgA response, and a local enhanced cellular response as demonstrated by lymphoproliferation of mesenteric lymph node cells upon in vitro stimulation with antigen. This immunization (120 micrograms in six oral doses at 2-day intervals) afforded mice a partial protection (60%) against a subsequent 400 oocyst challenge. The reduction in daily oocyst excretion was corroborated by significantly different weight losses between immunized and control mice on days 9 and 10 postinfection and the subsequent death of these control mice. These observations provide the first application of ISCOMs to parasitic intestinal diseases.

Keyword: immunotherapy

Failure of mycoplasma lipoprotein MALP-2 to induce NK cell activation through dendritic cell TLR2.

Macrophage-activating lipopeptide 2 (MALP-2), a mycoplasmal diacylated lipopeptide with moiety (Pam2), activates Toll-like receptor (TLR) 2 to induce inflammatory cytokines. TLR2 is known to mature myeloid dendritic cells (mDC) to drive mDC contact-mediated natural killer (NK) cell activation. Here we tested if MALP-2 activates NK cells through stimulation of TLR2 on mDC. Although synthetic MALP-2 with 6 or 14 amino acids (a.a.) stretch (designated as s and f) matured mDC to induce IL-6, IL-12p40 and TNF-α to a similar extent, they far less activated NK cells than Pam2CSK4, a positive control of 6 a.a.-containing diacyl lipopeptide. MALP-2s and f were TLR2/6 agonists and activate the MyD88 pathway similar to Pam2CSK4, but MALP-2s having the CGNNDE sequence acted on mDC TLR2 to barely induce external NK activation. Even the s form, with slightly high induction of IL-6 compared to the f form, barely induced in vivo growth retardation of NK-sensitive implant tumor. Pam2CSK4 and MALP-2 have the common lipid moiety but different peptides, which are crucial for NK cell activation. The results infer that MALP-2 is applicable to a cytokine inducer but not to an adjuvant for antitumor NK .Copyright © 2010 Institut Pasteur. Published by Elsevier SAS. All rights reserved.

Keyword: immunotherapy

Protein transfer enhances cellular immune responses to DNA vaccination against SARS-CoV.

The current DNA vaccine formulations are not optimal for stimulation of CD8(+) T cells, which are required for clearing virally-infected cells. Here we show that CD8(+) T cell-stimulating activity can be effectively augmented by combining DNA vaccination with protein transfer. C57BL/6 mice were injected intramuscularly with an anti-SARS-CoV DNA vaccine admixed with a lipid-derived conjugate of 4-1BBL, a potential CD8(+) T-cell co-stimulator. The inclusion of the lipidated co-stimulator greatly enhanced cellular immune responses, especially the CTL response, induced by the DNA vaccine. The adjuvant effect of 4-1BBL was lipidation-dependent, indicating that it functions as a cell membrane-anchored co-stimulator. Results of our study suggest, for the first time, that muscle cells may be modified in situ, at the DNA injection site, into APC-like cells to allow direct priming of CD8(+) T cells and thereby improve the efficacy of DNA vaccines.

Keyword: immunotherapy

Thioredoxin-interacting protein mediates hepatic lipogenesis and inflammation via PRMT1 and PGC-1α regulation in vitro and in vivo.

Non-alcoholic fatty liver disease (NAFLD) is strongly associated with obesity and type 2 diabetes. Thioredoxin-interacting protein (TXNIP) regulates the cellular redox state and metabolism and has been linked to many diseases, including diabetes. Therefore, we examined the role of TXNIP in hepatic steatosis in vitro and in vivo.Lipogenic and inflammatory proteins produced by hepatocytes treated with (PA) or transfected with TXNIP or Txnip siRNA were measured by Western blotting. Lipid accumulation was assessed using Oil Red O staining. Protein interactions were assessed by immunoprecipitation and proximity ligation assay. Hepatic protein levels were measured by Western blotting from wild type or Txnip(-/-) mice fed a high-fat diet (HFD) or chow diet. Livers from NAFLD patients were compared with normal liver by immunohistochemistry.PA increased TXNIP, and inflammatory and lipogenic proteins in both AML12 and H4IIE cells. It also increased the peroxisome proliferator-activated receptor gamma co-activator-1α (PGC-1α), which mediated the expression of lipogenic markers and lipid accumulation. In addition, PA increased protein arginine methyltransferase-1 (PRMT1) and PRMT1 siRNA abolished the increase in lipogenic markers with PGC-1α. Furthermore, TXNIP interacted with PRMT1 in PA-treated hepatocytes. In vivo, levels of lipogenic proteins, inflammatory molecules, PGC-1α, and PRMT1 were increased in the livers of HFD mice compared with those fed a chow diet, and were ameliorated in HFD Txnip(-/-) mice. Moreover, TXNIP, PRMT1, and PGC-1α were elevated in the livers of human NAFLD patients.TXNIP mediates hepatic lipogenesis via PRMT1 and PGC-1α regulation and inflammation in vitro and in vivo, implying that targeting TXNIP and PRMT1 is a potential therapeutic approach for treatment of NAFLD.Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: immunotherapy

Immunization of mice with human immunodeficiency virus glycoprotein gp160 peptide 315-329 induces both class I- and class II-restricted T cells: not all T cells can respond to whole molecule stimulation.

The V3 loop of human immunodeficiency virus (HIV) glycoprotein gp160 is of interest as a possible site for protective immune responses. This article examines the murine T cell response to peptide 315-329 derived from HIV gp160. Surprisingly, immunization with peptide in complete Freund\'s adjuvant induced class I-restricted T cells as well as class II-restricted T cells. These data suggest that this peptide may have the unusual ability to enter the class I antigen processing pathway. Strategies that employ V3 loop peptides to induce protective immunity must generate T cells that can recognize epitopes derived from whole molecules in vivo. Therefore, peptide-induced T cells were tested for their ability to respond to naturally processed forms of gp120 and gp160 whole-molecule preparations. Peptide induced class I-restricted cells were capable of recognizing transfectants expressing gp160. However, only one of two class II-restricted T cell lines was capable of recognizing soluble whole molecules. This indicates that peptide immunization induces T cells that recognize a class II-restricted determinant that is not generated during normal processing of whole molecules. We have also examined the response of peptide primed T cells to lipidated peptide antigens. Lipidated peptides are generally considered to have increased antigenicity and immunogenicity as compared to normal peptides. However, lipidation of peptide 315-329 damaged both the class I- and II-restricted determinants, indicating that lipidation is not always desirable. The data presented here highlight a potential serious problem in the use of peptide vaccines, in that peptide immunization may not always induce T cells that can protect against a viral challenge.

Keyword: immunotherapy

Synthesis and antigenicity of BBGL-2 glycolipids of Borrelia burgdorferi, the causative agent of Lyme disease.

Borrelia burgdorferi is the etiological agent for Lyme disease (LD), the most common vector borne disease in the United States. There is no human vaccine against LD currently available. Our approach to a vaccine is based on its surface-exposed glycolipids. One group of these glycolipids termed BBGL-2 consists of 1,2-di-O-acyl-3-O-(α-d-galactopyranosyl)-sn-glycerol congeners having , oleic, stearic, linoleic, and myristic acids. In order to delineate the immunodominant region(s) of the BBGL-2 components, we embarked on a synthetic project to provide available structurally defined, homogeneous analogs of BBGL-2 that might help identify the best vaccine candidate. The antigenicity of the synthetic glycolipids was examined by dot-blot analysis using mice sera obtained by immunization with killed B. burgdorferi cells, with native BBGL-2 in complete Freund\'s adjuvant, as well as sera obtained from patients with Lyme disease. We found that the presence of two acyl groups in the glycerol moiety was essential for antigenicity. At least one of these groups must be an oleoyl moiety. Neither the anomeric configuration of the galactose nor the configuration of the glycerol at C-2 was a decisive factor. Based on these findings we designed an \'unnatural\' BBGL-2 analog having the structure 3-O-(β-d-galactopyranosyl)-1,2-di-O-oleoyl-dl-glycerol which is easier and less expensive to synthesize than the other BBGL-2 congeners prepared in this study. This substance proved to be antigenic and is considered a candidate vaccine for Lyme disease.Published by Elsevier Ltd.

Keyword: immunotherapy

Palmitate-derivatized human IL-2: a potential anticancer immunotherapeutic of low systemic toxicity.

Recombinant human IL-2 (rhIL-2) is a potent cytokine and FDA-approved anticancer drug. However, its clinical use has been limited by severe toxicity, associated primarily with systemic administration with excess protein distributing freely throughout the body. We hypothesized that rhIL-2 in alternate forms permitting more restricted localization may exert stronger antitumor efficacy and less toxicity. Here, we have tested the utility of palmitate-derivatized rhIL-2. rhIL-2 was reacted with N-hydroxysuccinimide palmitate ester. The resultant lipidated rhIL-2 (pIL-2), when mixed with cells, could spontaneously transfer from solution to cell surfaces. Next, anticancer efficacy of pIL-2 was assessed in two modalities. For adoptive T cell therapy, antitumor cytotoxic T cells (CTLs) were protein transferred ("painted") with pIL-2 and injected into mice bearing lymphoma. For in situ therapy, pIL-2 was injected intratumorally into mice bearing melanoma. Tumor growth and IL-2-associated toxicity were determined.In the lymphoma model, painting of the antitumor CTLs with pIL-2 markedly increased their viability and titer. In the melanoma model, intratumoral injection of pIL-2, but not rhIL-2, increased the number of activated CD8(+) T cells (IFN-γ(+)) in the spleen, reduced lung metastasis and prolonged the survival of treated mice. Moreover, while repeated intratumoral injection of rhIL-2 at an excessively high dose (10 injections of 10,000\xa0IU/mouse) caused marked vascular leakage syndrome, the same regimen using pIL-2 caused no detectable toxicity.Transferring spontaneously from solution to cell surfaces, pIL-2 may bypass the current limitations of rhIL-2 and, thus, serve as a more effective and tolerable anticancer drug.

Keyword: immunotherapy

Immune response against the murine mdri protein induced by vaccination with synthetic lipopeptides in liposomes.

Intrinsically, or after exposure to chemotherapeutic drugs, many cancer cells overexpress a class of high molecular weight membrane glycoproteins associated with the multidrug resistance (mdr) of these cells. This report describes an immunization protocol eliciting autoantibodies specific to extracellular epitopes of the murine mdr 1 P-glycoprotein (Pgp). Synthetic peptides with the sequences of extracellular loops of murine Pgp were covalently coupled with four moieties per peptide molecule. These "lipopeptides" were reconstituted in the bilayer of liposomes containing lipid A and used to immunize mice. Antibodies against the lipopeptides corresponding to loop 2 and 4 were elicited in sera of immunized mice. They reacted specifically with extracellular epitopes of the naturally occurring murine Pgp. After interaction with resistant cancer cells, the antibodies induced an average 50% increase in cellular accumulation of doxorubicin and Bodipy-verapamil. In the presence of these antibodies the resistance of L1210 mdr cells was reduced from an LD50 of 4 x 10(-5) M to 5 x 10(-7) M doxorubicin.

Keyword: immunotherapy

Immune-spaying as an alternative to surgical spaying in Iberian×Duroc females: effect on carcass traits and meat quality characteristics.

The aim of this study was to assess the effect of immune-spaying on production traits and meat quality characteristics of fresh loin (longissimus dorsi) by comparing 3 groups of Iberian×Duroc females (N=12 per batch): surgically spayed, immune-spayed and entire females. Carcass traits and physicochemical parameters, including fatty profile, were investigated. The only carcass trait significantly affected by castration was the ham fat thickness, where both immune-spayed and surgically spayed females showed higher values against entire females (57±9.5mm, 62±2.5mm and 51±10.1mm, respectively; p<0.05). Furthermore, there were no significant differences in the quality parameters of fresh meat. These results are important regarding animal welfare, since in Europe, there is a plan to voluntarily end the surgical castration of pigs by 2018. Therefore, with an adequate vaccination protocol, immune-spaying might represent a good alternative to surgical spaying. The reliability of immune-spaying over long periods should be evaluated.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: immunotherapy

A synthetic peptide induces long-term protection from lethal infection with herpes simplex virus 2.

Immunization against viral pathogens is generally directed toward the induction of virus neutralizing antibody (VNA) and the maintenance of the potential for a second-set (IgG) response. Indeed, an elevated level of specific antibody is considered a reliable clinical indicator that a state of immunity exists in the host. However, in the case of herpes simplex virus (HSV), the presence of circulating VNA does not necessarily correlate with protection. Thus, it has been found that secondary infections occur in individuals even with high neutralizing titers to HSV, suggesting that antibody to the virus may be useless or even deleterious. In consideration of these facts, we were interested in inducing a T cell response to HSV. We had already shown that synthetic peptides corresponding to the NH3-terminal region of the glycoprotein D (gD) molecule of HSV could induce a strong T cell response when injected into mice, but did not, by themselves, confer protection. In this report, we examined the ability of peptides, covalently coupled to and incorporated into liposomes, to induce virus-specific T cell responses that confer protection against a lethal challenge of HSV-2. We have demonstrated that long-term protective immunity is achieved with a single immunization in the absence of neutralizing antibody when antigen is presented in this form. Furthermore, T cells but not serum from such immune mice can adoptively transfer this protection.

Keyword: immunotherapy

Long-term persistence of immunity induced by OVA-coupled gas-filled microbubble vaccination partially protects mice against infection by OVA-expressing Listeria.

Vaccination aims at generating memory immune responses able to protect individuals against pathogenic challenges over long periods of time. Subunit vaccine formulations based on safe, but poorly immunogenic, antigenic entities must be combined with adjuvant molecules to make them efficient against infections. We have previously shown that gas-filled microbubbles (MB) are potent antigen-delivery systems. This study compares the ability of various ovalbumin-associated MB (OVA-MB) formulations to induce antigen-specific memory immune responses and evaluates long-term protection toward bacterial infections. When initially testing dendritic cells reactivity to MB constituents, exhibited the highest degree of activation. Subcutaneous immunization of naïve wild-type mice with the OVA-MB formulation comprising the highest content and devoid of PEG2000 was found to trigger the more pronounced Th1-type response, as reflected by robust IFN-γ and IL-2 production. Both T cell and antibody responses persisted for at least 6 months after immunization. At that time, systemic infection with OVA-expressing Listeria monocytgenes was performed. Partial protection of vaccinated mice was demonstrated by reduction of the bacterial load in both the spleen and liver. We conclude that antigen-bound MB exhibit promising properties as a vaccine candidate ensuring prolonged maintenance of protective immunity.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: immunotherapy

MiRNA-194 Regulates -Induced Toll-Like Receptor 4 Inflammatory Responses in THP-1 Cells.

There is strong evidence to suggest that inflammatory responses link obesity and diseases, and the understanding of obesity-induced inflammatory mechanisms is central to the pathogenesis of diseases such asnonalcoholic fatty liver disease(NAFLD) and atherosclerosis that are modified by obesity. Based on this, anti-inflammatory treatments become a potential therapies for obesity-related diseases like NAFLD.A critical role of toll-like receptor (TLR) and its downstream molecules such as tumor necrosis factor receptor-associated factor 6(TRAF6) has been documented in inflammatory response induced by fatty . TLR pathway regulation provides a new insight to controlling the inflammatory response induced by fatty . Taken together, our study was aimed to understand the mechanism of fatty -mediated inflammation and look for an effective target which can prevent the inflammatory response induced by obesity. In this study, we used the saturated fatty (PA) to activate TLR4 signal pathway in human monocyte cells THP-1 that established an intracellular inflammatory model. Followed with activated TLR4, downstream molecular TRAF6 was upregulated and ultimately induced proinflammatory cytokine production. Based on this model, we also found that PA downregulated miR-194 expression with TLR4 activation. Moreover, our results showed that key signal molecular TRAF6 is a target of miR-194, overexpression of miR-194 directly decreased TRAF6 expression and attenuated the release of proinflammatory cytokine TNF-α in PA-activated monocyte THP-1. We conclude that miR-194 negatively regulates the TLR4 signal pathway which is activated by PA through directly negative TRAF6 expression.

Keyword: immunotherapy

Efficacy of a therapeutic treatment using gas-filled microbubble-associated phospholipase A2 in a mouse model of honeybee venom allergy.

Venom is efficient to desensitize people suffering from insect sting allergies. However, the numerous injections required over several years and important risks of severe side reactions complicate the widespread use of . In the search for novel approaches to blunt the overwhelming pro-allergic Th2 response, we evaluated the therapeutic efficacy of a treatment based on a denatured form of the major allergen, phospholipase A2, associated with microbubbles (PLA2denat -MB) in a mouse model of honeybee venom allergy.Antibodies measured by ELISA, T-cell responses assessed by CFSE-based proliferation assays and ELISA, and basophil degranulation were examined\xa0after PLA2denat -MB-based therapeutic treatment of sensitized mice. Mice were challenged with a lethal dose of PLA2 to evaluate protection against anaphylaxis.Therapeutic subcutaneous administration of two different PLA2denat -MB formulations, in contrast to PLA2denat alone, reduced allergic symptoms and protected all mice from anaphylaxis-mediated death after allergen challenge. At the functional level, the use of PLA2denat decreased IgE-mediated basophil degranulation as compared to the native form of the allergen. In comparison with PLA2denat alone, both PLA2denat -MB formulations decreased allergen-specific Th2 CD4 T-cell reactivity. At the mechanistic level, PLA2denat -MB containing 20% and PEG induced PLA2-specific IgA and increased Foxp3(+) Treg frequencies and TGF-β production, whereas the formulation bearing 80% triggered the production of IFN-γ, IgG2a, and IgG3.In contrast to conventional PLA2 subcutaneous , the therapeutic administration of PLA2-MB treatment to mice that already had established allergy to PLA2 protects all subsequently challenged animals.© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: immunotherapy

A recombinant non-fatty acylated form of the Hi-PAL (P6) protein of Haemophilus influenzae elicits biologically active antibody against both nontypeable and type b H. influenzae.

An approximately 15,000-dalton outer membrane lipoprotein of Haemophilus influenzae, the Hi-PAL (P6) protein, has been shown to elicit bactericidal and protective antibodies against both type b and nontypeable H. influenzae strains and is a vaccine candidate for these organisms. To determine whether the lipid modification of this protein is required for immunogenicity or the elicitation of biologically active antibodies, a genetic fusion was constructed that contains the sequence of mature Hi-PAL fused to the polylinker region of pUC19. The protein expressed by this clone does not contain detectable lipid and was purified to homogeneity. This recombinant fusion protein, rPAL, elicited a strong immune response when injected into rabbits, and the antiserum reacted well with native Hi-PAL. The antiserum was bactericidal against a number of clinical nontypeable strains, duplicating the activity of anti-Hi-PAL. The anti-rPAL antiserum was also protective against type b bacteremia in the infant rat model. These results demonstrate that purified rPAL elicits antibodies with biological activities that are similar to those of anti-Hi-PAL antibodies. Thus, the lipid component of Hi-PAL is not required for either immunogenicity or elicitation of biologically active antibodies.

Keyword: immunotherapy

Preparation of immune stimulating complexes (ISCOMs) as adjuvants.

Purified proteins are often poorly immunogenic and in such cases the induction of primary immune responses requires use of an adjuvant. The immune stimulating complex (ISCOM) has a unique ability to provoke a full range of immune response to protein antigens, after both parenteral and oral immunization. This unit describes techniques for incorporating proteins into the ISCOM structure, a process that requires the presence of exposed hydrophobic regions on the protein. The basic protocol outlines a method for preparation of ISCOMs containing inherently nonhydrophobic proteins, to which has been attached covalently. Two alternate protocols are given that do not require covalent modification of the protein. In the first, hydrophobic groups are revealed by treatment of the protein. The second describes preparation of ISCOMs containing integral membrane proteins that therefore possess a hydrophobic transmembrane domain.

Keyword: immunotherapy

Ingested interferon alpha suppresses type I diabetes in non-obese diabetic mice.

Type I diabetes mellitus is a chronic disorder that results from autoimmune destruction of the insulin-producing pancreatic beta cell. The non-obese diabetic mouse is a model of the human autoimmune disease Type I diabetes [1-3]. We have previously shown that ingested type 1 interferon inhibits chronic relapsing experimental autoimmune encephalomyelitis and the adoptive transfer of experimental autoimmune encephalomyelites by T cells, and decreases both antigen-specific and mitogen-induced pro-inflammatory cytokine secretion in this disorder. We therefore tried to determine whether ingested murine interferon alpha inhibits insulinitis and suppresses Type I diabetes mellitus in non-obese diabetic mice. Murine interferon alpha, given daily, decreased islet inflammation and suppressed diabetes. It increased the concanavalin A and ionomycin plus myristic ester-induced production of interleukin 4 and 10 and interferon gamma-secretion in spleen cells from treated mice. Adoptive transfer of unstimulated splenocytes secreting interleukin 4 and interleukin 10 from fed interferon alpha donors suppressed spontaneous diabetes mellitus in recipients. The protective effect of adoptively transferred unstimulated splenocytes shows the presence of ingested interferon alpha-activated regulatory splenic cell populations that may work via increased interleukin 4 or interleukin 10 production. Ingested interferon alpha administered during vulnerable periods in at-risk populations may potentially provide a continuous, convenient, non-toxic and effective treatment for Type I diabetes.

Keyword: immunotherapy

The effect of endotoxin on membrane fatty composition in BCG-sensitized mice.

The effects of endotoxin on mouse liver phospholipid fatty composition have been investigated. Administration of endotoxin from Salmonella abortus equi led to a decrease in the polyunsaturated fatty content of livers from mice sensitized with Bacille Calmette Guérin (BCG). The content of arachidonic fell significantly in both the phosphatidylcholine and phosphatidylinositol fractions whereas in the phosphatidylethanolamine fraction the linoleic content was significantly reduced. The polyunsaturated fatty acids were replaced by increased amounts of oleic and , leading to a reduction in the polyunsaturated to saturated fatty ratio.

Keyword: immunotherapy

Linear and branched glyco-lipopeptide vaccines follow distinct cross-presentation pathways and generate different magnitudes of antitumor immunity.

Glyco-lipopeptides, a form of lipid-tailed glyco-peptide, are currently under intense investigation as B- and T-cell based vaccine for many cancers. However, the cellular and molecular mechanisms of glyco-lipopeptides (GLPs) immunogenicity and the position of the lipid moiety on immunogenicity and protective efficacy of GLPs remain to be determined.We have constructed two structural analogues of HER-2 glyco-lipopeptide (HER-GLP) by synthesizing a chimeric peptide made of one universal CD4(+) epitope (PADRE) and one HER-2 CD8(+) T-cell epitope (HER(420-429)). The C-terminal end of the resulting CD4-CD8 chimeric peptide was coupled to a tumor carbohydrate B-cell epitope, based on a regioselectively addressable functionalized templates (RAFT), made of four alpha-GalNAc molecules. The resulting HER glyco-peptide (HER-GP) was then linked to a moiety, attached either at the N-terminal end (linear HER-GLP-1) or in the middle between the CD4+ and CD8+ T cell epitopes (branched HER-GLP-2). We have investigated the uptake, processing and cross-presentation pathways of the two HER-GLP vaccine constructs, and assessed whether the position of linkage of the lipid moiety would affect the B- and T-cell immunogenicity and protective efficacy. Immunization of mice revealed that the linear HER-GLP-1 induced a stronger and longer lasting HER(420-429)-specific IFN-gamma producing CD8(+) T cell response, while the branched HER-GLP-2 induced a stronger tumor-specific IgG response. The linear HER-GLP-1 was taken up easily by dendritic cells (DCs), induced stronger DCs maturation and produced a potent TLR- 2-dependent T-cell activation. The linear and branched HER-GLP molecules appeared to follow two different cross-presentation pathways. While regression of established tumors was induced by both linear HER-GLP-1 and branched HER-GLP-2, the inhibition of tumor growth was significantly higher in HER-GLP-1 immunized mice (p<0.005).These findings have important implications for the development of effective GLP based immunotherapeutic strategies against cancers.

Keyword: immunotherapy

A pilot study of the CY-1899 T-cell vaccine in subjects chronically infected with hepatitis B virus. The CY1899 T Cell Vaccine Study Group.

Clinical observations suggest that eradication of the hepatitis B virus (HBV) is immune-mediated. Vigorous cytotoxic T lymphocyte (CTL) activity directed at HLA class I-bound viral epitopes are detected during acute hepatitis B, but not in chronic hepatitis B carriers. A CTL epitope derived from the hepatitis B core protein amino acids 18-27 has been incorporated into a vaccine also comprised of a T-helper cell epitope and 2 residues (CY-1899). The aim of this study was to determine whether repeated doses of CY-1899 given to patients with chronic hepatitis B could initiate in vivo CTL activity and viral clearance. Patients with chronic hepatitis B received up to 4 doses (ranging from 0.05 mg to 15 mg) 6 weeks apart. Following vaccination, patients were monitored for hepatitis B surface antigen and "e" status, HBV-DNA levels, liver biochemistry, CTL activity, and any adverse events. Ninety patients with chronic hepatitis B infection received CY-1899. Mean CTL responses were all low but were maximal following vaccination with 5 mg CY-1899. Peak CTL responses never exceeded 10 lytic units (LU) regardless of vaccine dose, this value being well below that seen following resolution of acute hepatitis B. No significant changes in liver biochemistry or viral serology were observed during follow-up. No serious adverse events were noted. Administration of the single-epitope vaccine, CY-1899, initiated CTL activity, but of a magnitude lower than that observed during spontaneous HBV clearance. This low-level CTL activity was not associated with viral clearance.

Keyword: immunotherapy

A protective glycosylphosphatidylinositol-anchored membrane protein of Plasmodium yoelii trophozoites and merozoites contains two epidermal growth factor-like domains.

Using sera from mice immunized and protected against Plasmodium yoelii malaria, we identified a novel blood-stage antigen gene, pypag-2. The 2.1-kb pypag-2 cDNA contains a single open reading frame that encodes a 409-amino- protein with a predicted molecular mass of 46.8 kDa. Unlike many characterized plasmodial antigens, blocks of tandemly repeated amino acids are lacking in the pypAg-2 protein sequence. Recombinant pypAg-2, comprising the full-length protein minus the predicted N-terminal signal and C-terminal anchor sequences, was produced and used to raise a high-titer polyclonal rabbit antiserum. This antiserum was used to identify and characterize the native protein through immunoblotting, immunoprecipitation and immunofluorescence assays. Consistent with the presence of a glycosylphosphatidylinositol anchor, pypAg-2 fractionated with the detergent phase of Triton X-114-solubilized proteins and could be metabolically labeled with [(3)H]. By immunofluorescence, pypAg-2 expression was localized to both the trophozoite and merozoite membranes. Similar to Plasmodium falciparum merozoite surface protein 1, pypAg-2 contains two C-terminal epidermal growth factor (EGF)-like domains. Most importantly, immunization with recombinant pypAg-2 protected mice against lethal P. yoelii malaria. Thus, pypAg-2 is a target of protective immune responses and represents a novel addition to the family of merozoite surface proteins that contain one or more EGF-like domains.

Keyword: immunotherapy

Osmotic shock fluids from salmonella R-mutants. Chemical composition and protective capacity against S. typhimurium infection in mice.

Salmonella R-mutants of different chemotype were subjected to osmotic shock treatment according to the method of Willis et al. (18). Chemical composition (protein, LPS, and nucleic acids) as well as the polypeptide patterns of the shock fluid and hypertonic fluid (in which the cells were suspended before shock treatment) were determined and compared with material extracted by urea treatment carried out to get protein from the cell surface. The results indicate that even components from the cell surface have been released by shock treatment. All the extracts mentioned above showed protective activity in mouse against infection.

Keyword: immunotherapy

Synthetic peptide vaccines: palmitoylation of peptide antigens by a thioester bond increases immunogenicity.

Synthetic peptides have frequently been used to immunize animals. However, peptides less than about 20 to 30 amino acids long are poor immunogens. In general, to increase its immunogenicity, the presentation of the peptide should be improved, and molecular weight needs to be increased. Many attempts have been made to couple peptide immunogens to different carrier proteins [e.g. keyhole limpet haemocyanin (KLH) or ovalbumin]. This leads to very complex structures, however. We used a controlled conjugation of a peptide to a single long-chain fatty like by a thioester or an amide bond. It was found that these S-palmitoylated peptides were much more immunogenic than N-palmitoylated peptides and at least similar to KLH-conjugated peptides with respect to appearance and magnitude of induced antibodies (canine parvovirus) or immunocastration effect (gonadotropin-releasing hormone). For chemical synthesis of thioesters, we established conditions for solution and solid-phase synthesis. In both phases, Cys(SBut) could only be deprotected efficiently using phosphines, and S-acylation was accomplished using standard coupling at pH 5. We speculate that, in vivo, the presence of an appropriate fatty chain, chemically linked through a labile thioester bond, greatly enhances immunogenicity, because it represents a favourable substrate for cleavage by cellular thioesterases in cells of the immune system.

Keyword: immunotherapy

Fully synthetic carbohydrate-based vaccines in biochemically relapsed prostate cancer: clinical trial results with alpha-N-acetylgalactosamine-O-serine/threonine conjugate vaccine.

We report the synthesis of a mucin-related O-linked glycopeptide, alpha-N-acetylgalactosamine-O-serine/threonine (Tn), which is highly simplistic in its structure and can induce a relevant humoral response when given in a trimer or clustered (c) formation. We tested for an antitumor effect, in the form of a change in the posttreatment versus pretreatment prostate-specific antigen (PSA) slopes, that might serve as a surrogate for effectiveness of vaccines in delaying the time to radiographic progression.We compared the antibody response to immunization with two conjugates, Tn(c)-keyhole limpet hemocyanin (KLH) and Tn(c)- (PAM) with the saponin immunologic adjuvant QS21, in a phase I clinical trial in patients with biochemically relapsed prostate cancer. Patients received Tn(c)-KLH vaccine containing either 3, 7, or 15 microg of Tn(c) per vaccination. Ten patients received 100 microg of Tn(c)-PAM. QS21 was included in all vaccines. Five vaccinations were administered subcutaneously during 26 weeks with an additional booster vaccine at week 50.Tn(c), when given with the carrier molecule KLH and QS21, stimulated the production of high-titer immunoglobulin M (IgM) and IgG antibodies. Inferior antibody responses were seen with T(c)-PAM. There was no evidence of enhanced immunogenicity with increasing doses of vaccine. An antitumor effect in the form of a decline in posttreatment versus pretreatment PSA slopes was also observed.A safe synthetic conjugate vaccine in a trimer formation was developed that can break immunologic tolerance by inducing specific humoral responses. It seemed to affect the biochemical progression of the disease as determined by a change in PSA log slope.

Keyword: immunotherapy

Tolerance induction by acylated peptides: suppression of EAE in the mouse with palmitoylated PLP peptides.

Treatment of SJL mice either before or after challenge with palmitoylated PLP139-151 (PAL139-151) completely suppressed or considerably reduced both acute and relapsing stages of EAE induced with PLP139-151. In the presence of Pertussis toxin, treatment with PAL139-151 was less effective, but treatment with a mixture of PAL139-151 and PAL178-191, the palmitoylated PLP epitope to which T cell recognition spreads, resulted in almost complete protection. Proliferation of lymphocytes from treated mice were sharply reduced, and adoptive transfer of lymph node lymphocytes from treated mice to naive recipients resulted in the reduction of the acute phase of EAE and in delayed relapses following challenge. The results suggest that treatment with PAL139-151 leads to both anergy and the generation of regulatory cells.

Keyword: immunotherapy

Suppression of IgE antibody response by the fatty -modified antigen.

Ovalbumin (OA) of hens was chemically coupled with fatty acids (lauric , myristic , and stearic ). These hydrophobically modified antigens were unable to react with mouse antiserum against native OA and were incapable of eliciting primary and secondary anti-OA antibody responses in BALB/c mice. Preadministration of these modified antigens, especially of palmitoyl OA (OA-pal), suppressed both primary and secondary anti-OA IgE antibody responses without affecting IgG antibody production. Administration of OA-pal after the primary immunization resulted in a rapid decrease of the ongoing anti-OA IgE antibody production and inhibited the anamnestic anti-OA IgE antibody response upon subsequent immunization with OA. The passive transfer of spleen cells from OA-pal-treated animals with OA-primed spleen cells suppressed the adoptive secondary anti-OA IgE antibody response in irradiated recipients. The suppressive effect was abrogated by treatment with an anti-T-cell antiserum indicating that suppressor T cells were primed by administration of hydrophobically modified antigens.

Keyword: immunotherapy

Phytochemistry, Traditional Uses and Pharmacological Profile of Rose Hip: A Review.

The fruit of genus Rosa, known as "rose hip", is frequently used in different traditional medicines. Rose hips have long been used to treat kidney stones, gastroenteric ailments, hypertension and respiratory problems such as bronchitis, cough and cold.This review is focused on the ethnopharmacological uses of rose hip as well as phytochemical and pharmacological aspects.Ethno-medical uses of rose hip have been recorded in many countries since a long time. Approximately, 129 chemical compounds have been isolated and identified from rose hip. This fruit contains some major active components such as flavonoids, tannins, anthocyanin, phenolic compounds, fatty oil, organic acids and inorganic compounds. Scientific studies have suggested a wide range of pharmacological activities for rose hip including antioxidant, anti-inflammatory, anti-obesity, anti-cancer, hepatoprotective, nephroprotective, cardioprotective, antiaging, anti H. pylori, neuroprotective and antinociceptive activities. In particular, the rose hip powder and extract have been reported to exert therapeutic effects on arthritis.Some of the ethnomedical indications of rose hip, such as nephroprotective and gastroproetctive actions, have been confirmed by preclinical pharmacological studies. Additional investigations on the pharmacological effects of rose hip as well as evidence from randomized controlled trials are essential to assess the therapeutic value of this natural product.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: inflammation

[Protective effects of total saponins of Panax japonicas on HepG2 cell apoptosis induced with ].

This study aimed to investigate the molecular mechanism and protective effect of total saponins of Panax japonicas (TSPJ) on HepG2 cells apoptosis induced by (PA).The HepG2 cells were cultured , and divided into five groups: the control group, the model group, the high-dose group (50 mg·L⁻¹), the middle-dose group (25 mg·L⁻¹) and the low-dose group (12.5 mg·L⁻¹).The cells of the five groups were cultured continuously for 24 hours. The cell viability was measured with MTT. HepG2 cells apoptosis was detected by Hoechest staining and Annexin V-FITC/PI staining. The protein expressions of BCL-2, CHOP and TLR4 were measured with western blotting and flow cytometry analysis. The mRNA expressions of TNF-α, IL-1β, BCL-2, CHOP and GAPDH were measured with RT-PCR. The results suggested that compared with the control group, the number of HepG2 cells of the model group were reduced significantly (<0.01), while the number of apoptotic HepG2 cells were increased. Compared with the model group, the number of HepG2 cells of the high-dose group and the middle-dose group were increased significantly (<0.01), whereas the number of apoptotic HepG2 cells were reduced. Compared with the control group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the model group were significantly up-regulated (<0.01), while BCL-2 protein and mRNA expressions in the model group were significantly decreased (<0.01). Compared with the model group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the high-dose group were significantly decreased (<0.01), while BCL-2 protein and mRNA expressions in the high-dose group were significantly up-regulated (<0.01).In conclusion, TSPJ can reduce and apoptosis induced by , with a certain protective effect on liver cells.Copyright© by the Chinese Pharmaceutical Association.

Keyword: inflammation

Omega-3 Polyunsaturated Fatty Acids Inhibit IL-17A Secretion through Decreased ICAM-1 Expression in T\xa0Cells Co-Cultured with Adipose-Derived Stem Cells Harvested from Adipose Tissues of Obese Subjects.

Obese adipose tissue (AT) is infiltrated by inflammatory immune cells including IL-17A-producing-T (Th17) cells. It has been previously demonstrated that adipose-derived stem cells from obese (ob-ASCs), but not lean AT promote Th17 cells. Because n-3 PUFAs are known to inhibit obese AT , it is tested here whether they could inhibit ob-ASC-mediated IL-17A secretion.The n-3 PUFA precursor, alpha-linolenic (ALA), or its derivatives, eicosapentaenoic, or docosahexaenoic , is added to co-cultures of human ob-ASCs and mononuclear cells (MNCs). All three inhibited IL-17A, but not IL-1β, IL-6, nor TNFα\xa0 secretion. As a control, (PA), a saturated fatty , did not inhibit IL-17A secretion. ALA also inhibited IL-17A secretion mediated by adipocytes differentiated from ob-ASCs. Toll-like-receptor 4 is shown to be involved in ob-ASC-mediated-IL-17A secretion, and to be inhibited by ALA, together with Cyclo-Oxygenase-2 and Signal-Transducer-and-Activator-of-transcription-3. In addition, ALA down-regulated Intercellular-Adhesion-Molecule-1 (ICAM-1) expression in both monocytes and ASCs, which resulted in decreased interactions between ob-ASCs and MNCs, and inhibition of IL-17A secretion.It is demonstrated herein that ALA inhibits Th17 cell promotion, through decreased ICAM-1expression in both ob-ASCs and monocytes. This novel mechanism may contribute to explain the beneficial effects of n-3 PUFA in IL-17A-related inflammatory pathologies.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: inflammation

Is palmitate truly proinflammatory? Experimental confounders and context-specificity.

Based primarily on cell culture results, saturated fatty acids (SFAs) are proposed to promote and contribute to metabolic dysfunction through Toll-like receptor activation. Studies are often complicated by a requirement for carriers (e.g., BSA) or solvents (e.g., ethanol) to increase SFA solubility. To ascertain whether these factors influence interpretations of SFA-associated activity, we measured responses of RAW264.7 monocyte/macrophages and CC myotubes to various BSA, ethanol, and cyclodextrin (alternative FA carrier) conditions. Fatty -free, low-endotoxin BSA preparations (0.33% to 2% wt/vol) activated whereas 0.5-1.0% ethanol inhibited RAW264.7 TNFα release. Ethanol modestly increased IL-6 secretion in CC myotubes. Cyclodextrins (0.3-6.0 mM) were tested as alternative carriers of palmitate, but their usefulness was limited due to toxicity and solubility issues. Using a lower- BSA source and no ethanol, ∼24-h sodium palmitate treatment (≤600 µM) failed to trigger RAW264.7 TNFα release and, in fact, significantly dampened BSA-induced by >50%. In CC myotubes, only high palmitate concentrations (500-600 µM) elicited IL-6 secretion (>2.5-fold increase). Acute palmitate (200 or 500 µM) treatment did not activate MAP kinase pathways above that of fresh BSA-containing media alone in either cell type. These results highlight the importance of experimental conditions in studies exploring SFA effects. The limited (or even anti-inflammatory) effects of palmitate that we observed indicate that immunomodulatory effects of SFAs are context-specific. Thus, caution is needed when interpreting the literature related to putative proinflammatory effects of SFA.

Keyword: inflammation

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and -suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on lipid metabolism and in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular lipid contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet lipid metabolic disorder and islet in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: inflammation

Ginsenoside Rg1 Ameliorates -Induced Hepatic Steatosis and in HepG2 Cells via the AMPK/NF-B Pathway.

Nonalcoholic fatty liver disease (NAFLD) is one of the common diseases in the world, and it can progress from simple lipid accumulation to sustained . The present study was designed to investigate the effects and underlying mechanisms of ginsenoside Rg1 (G-Rg1) treatment on NAFLD . HepG2 cells were treated with (PA) to induce steatosis and and then successively incubated with G-Rg1. Lipids accumulation was analyzed by Oil Red O staining and intracellular triglyceride (TG) quantification. Inflammatory conditions were examined by quantifying the levels of cell supernatant alanine transaminase/aspartate aminotransferase (ALT/AST) and secretory proinflammatory cytokines, including IL-1, IL-6, and TNF- in the cell supernatants. Quantitative RT-PCR and western blotting were used to measure the expressions of genes and proteins associated with lipogenic synthesis and , including AMP-activated protein kinase (AMPK) and nuclear factor-kappa B (NF-B) pathways. HepG2 cells were pretreated with an AMPK inhibitor; then, Oil Red O staining and TG quantification were performed to study the lipid deposition. Phospho-AMPK (Thr172) (p-AMPK) and phospho-acetyl-CoA carboxylase (Ser79) (p-ACC) were quantified by immunoblotting. Immunofluorescence was performed to demonstrate the nuclear translocation of NF-B P65. The present study showed that PA markedly increased the intracellular lipid droplets accumulation and TG levels, but decreased AMPK phosphorylation and the expressions of its downstream lipogenic genes. However, G-Rg1 alleviated hepatic steatosis and reduced the intracellular TG content; these changes were accompanied by the activation of the AMPK pathway. In addition, blocking AMPK by using the AMPK inhibitor markedly abolished the G-Rg1-mediated protection against PA-induced lipid deposition in HepG2 cells. Furthermore, G-Rg1 reduced the ALT/AST levels and proinflammatory cytokines release, which were all enhanced by PA. These effects were correlated with the inactivation of the NF-B pathway and translocation of P65 from the cytoplasm to the nucleus. Overall, these results suggest that G-Rg1 effectively ameliorates hepatic steatosis and , which might be associated with the AMPK/NF-B pathway.

Keyword: inflammation

Preliminary evidence for reduced adipose tissue inflammation in vegetarians compared with omnivores.

There are links between obesity and inflammation that may relate activation of pro-inflammatory pathways by dietary factors. Because dietary fat intake of vegetarians is thought to be more beneficial than that of omnivores, we hypothesized that obese vegetarians would have less adipose tissue inflammation and lower intramyocellular ceramide concentrations than equally obese omnivores.Eight obese vegetarian (1 male) and 8 obese omnivore volunteers (1 male) completed a Food Frequency Questionnaire, underwent body composition measures, subcutaneous adipose tissue and muscle biopsies. We used immunohistochemistry to measure adipose macrophage (ATM) and senescent cells. Plasma free fatty (FFA), adipose FA and muscle ceramide profiles were measured using liquid chromatography/mass spectrometry. Student t tests were used for the comparison of primary outcomes; univariate regression analysis was used to test for associations between dietary patterns and ATMs (secondary analysis).There were no differences in age (38\u2009±\u20098 vs. 39\u2009±\u20098\u2009years), BMI (32.2\u2009±\u20092.6 vs. 33.3\u2009±\u20091.9\u2009kg/m) or percent body fat (44\u2009±\u20098 vs. 45\u2009±\u20094) between the vegetarians and omnivores. Vegetarians consumed 42% (P\u2009=\u20090.02) less saturated fat and 50% (P\u2009=\u20090.04) less cholesterol than the omnivores. Plasma FFA of vegetarians had lesser proportions of (24\u2009±\u20093 vs. 29\u2009±\u20094%, P\u2009=\u20090.02) and vegetarians had fewer femoral pro-inflammatory ATMs than omnivores (3.6\u2009±\u20092.8 vs. 7.9\u2009±\u20094.4 per 100 adipocytes, respectively; P\u2009=\u20090.02). Omnivores had 50% greater (P\u2009=\u20090.01) expression of TNF mRNA in abdominal fat. We found no significant between group differences in muscle ceramide concentrations.Although the sample size is small, these results may indicate that dietary patterns play a role in adipose tissue inflammation, as reflected by reduced number of femoral ATMs in obese vegetarians than obese omnivores.

Keyword: inflammation

Cytotoxic effect of Kalanchoe flammea and induction of intrinsic mitochondrial apoptotic signaling in prostate cancer cells.

Kalanchoe flammea Stapf (Crassulaceae) is a medicinal plant grown in the South of Mexico (State of Tabasco), which is commonly used in traditional medicine for the treatment of fever, wounds, , and cancer.To establish the potential of K. flammea for the treatment of prostate cancer, evaluating its cytotoxic activity, its probable mechanism of action, and carrying out some toxicological safety studies.The cytotoxic activity of the ethyl acetate extract of K. flammea (Kf-EtOAc) was evaluated in several cell lines of prostate cancer by MTT viability assay. The cellular death mechanism was studied by evaluating the translocation of phosphatidylserine (Annexin V); overproduction of reactive oxygen species [2\'-7\'-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay]; release of Cytochrome C; activation of caspase-3 and\xa0-9, and regulation of Bcl-2, XIAP, and PKCε proteins by Western Blot analysis. For the evaluation of the safety of Kf-EtOAc, the Ames test, Micronucleus assay, and acute toxicity study were determined.Kf-EtOAc exhibited selective cytotoxic activity against prostate cell lines as follows: PC-3, LNCaP, and PrEC (IC = 1.36\u202f±\u202f0.05; 2.06\u202f±\u202f0.02, and 127.05\u202f±\u202f0.07\u202fμg/mL, respectively). The F82-P2 fraction (rich in coumaric and ) obtained by bioassay-guided fractionation of Kf-EtOAc also demonstrated selective cytotoxic activity against PC-3 cells (IC = 1.05\u202f±\u202f0.06\u202fμg/mL). Kf-EtOAc induces apoptosis by the intrinsic pathway; this mechanism of cell death was confirmed after observing that the extract produces phosphatidylserine translocation, overproduction of reactive oxygen species, release of Cytochrome C at mitochondrial level, and activation of caspase-3 and -9. It was also observed that Kf-EtOAc produces significant downregulation of apoptosis-related proteins Bcl-2, XIAP, and PKCε and induces DNA fragmentation and cell cycle arrest. In addition, Kf-EtOAc is non-genotoxic in vitro by Ames test and non-genotoxic in vivo by Micronucleus assay, and no signs of toxicity or death were reported after the administration of a single acute exposure of 2000\u202fmg/kg.K. flammea is a potential candidate for the development of new drugs for the treatment of prostate cancer. However, to propose their use in clinical trials, additional studies are required to understand their pharmacokinetic behavior, as well as the development of a suitable pharmaceutical form.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

Palmitoylethanolamide Promotes a Proresolving Macrophage Phenotype and Attenuates Atherosclerotic Plaque Formation.

Objective- Palmitoylethanolamide is an endogenous fatty mediator that is synthetized from membrane phospholipids by N-acyl phosphatidylethanolamine phospholipase D. Its biological actions are primarily mediated by PPAR-α (peroxisome proliferator-activated receptors α) and the orphan receptor GPR55. Palmitoylethanolamide exerts potent anti-inflammatory actions but its physiological role and promise as a therapeutic agent in chronic arterial , such as atherosclerosis remain unexplored. Approach and Results- First, the polarization of mouse primary macrophages towards a proinflammatory phenotype was found to reduce N-acyl phosphatidylethanolamine phospholipase D expression and palmitoylethanolamide bioavailability. N-acyl phosphatidylethanolamine phospholipase D expression was progressively downregulated in the aorta of apolipoprotein E deficient (ApoE) mice during atherogenesis. N-acyl phosphatidylethanolamine phospholipase D mRNA levels were also downregulated in unstable human plaques and they positively associated with smooth muscle cell markers and negatively with macrophage markers. Second, ApoE mice were fed a high-fat diet for 4 or 16 weeks and treated with either vehicle or palmitoylethanolamide (3 mg/kg per day, 4 weeks) to study the effects of palmitoylethanolamide on early established and pre-established atherosclerosis. Palmitoylethanolamide treatment reduced plaque size in early atherosclerosis, whereas in pre-established atherosclerosis, palmitoylethanolamide promoted signs of plaque stability as evidenced by reduced macrophage accumulation and necrotic core size, increased collagen deposition and downregulation of M1-type macrophage markers. Mechanistically, we found that palmitoylethanolamide, by activating GPR55, increases the expression of the phagocytosis receptor MerTK (proto-oncogene tyrosine-protein kinase MER) and enhances macrophage efferocytosis, indicative of proresolving properties. Conclusions- The present study demonstrates that palmitoylethanolamide protects against atherosclerosis by promoting an anti-inflammatory and proresolving phenotype of lesional macrophages, representing a new therapeutic approach to resolve arterial .

Keyword: inflammation

The Distinct Effects of and Oleic on Pancreatic Beta Cell Function: The Elucidation of Associated Mechanisms and Effector Molecules.

In this study, we aimed to identify the mechanisms underlying the different effects of and oleic on human pancreatic beta cell function. To address this problem, the oxidative stress, endoplasmic reticulum stress, , apoptosis and their mediator molecules have been investigated in the insulin releasing beta cells exposed to and/or oleic . Herein, we have demonstrated that in cultured 1.1B4 beta cells oleic promotes neutral lipid accumulation and insulin secretion, whereas is poorly incorporated into triglyceride and it does not stimulate insulin secretion from human pancreatic islets at physiologically glucose concentrations. In addition, caused: oxidative stress through a mechanism involving increases in ROS production and MMP-2 protein expression/gelatinolytic activity associated with down-regulation of SOD2 protein; endoplasmic reticulum stress by up-regulation of chaperone BiP protein and unfolded protein response (UPR) transcription factors (eIF2α, ATF6, XBP1u proteins) and by PTP-1B down-regulation in both mRNA and protein levels; through enhanced synthesis of proinflammatory cytokines (IL6, IL8 proteins); and apoptosis by enforced proteic expression of CHOP multifunctional transcription factor. Oleic alone had opposite effects due to its different capacity of controlling these metabolic pathways, in particular by reduction of the ROS levels and MMP-2 activity, down-regulation of BiP, eIF2α, ATF6, XBP1u, CHOP, IL6, IL8 and by SOD2 and PTP-1B overexpression. The supplementation of saturated with the monounsaturated oleic reversed the negative effects of alone regulating insulin secretion from pancreatic beta cells through ROS, MMP-2, ATF6, XBP1u, IL8 reduction and SOD2, PTP-1B activation. Our findings have shown the protective action of oleic against on beta cell lipotoxicity through promotion of triglyceride accumulation and insulin secretion and regulation of some effector molecules involved in oxidative stress, endoplasmic reticulum stress, and apoptosis.

Keyword: inflammation

Multi-omics analysis points to altered platelet functions in severe food-associated respiratory allergy.

Prevalence and severity of allergic diseases have increased worldwide. To date, respiratory allergy phenotypes are not fully characterized and, along with progression, treatment is increasingly complex and expensive. Profilin sensitization constitutes a good model to study the progression of allergic . Our aim was to identify the underlying mechanisms and the associated biomarkers of this progression, focusing on severe phenotypes, using transcriptomics and metabolomics.Twenty-five subjects were included in the study. Plasma samples were analyzed using gas and liquid chromatography coupled to mass spectrometry (GC-MS and LC-MS, respectively). Individuals were classified in four groups-"nonallergic," "mild," "moderate," and "severe"-based on their clinical history, their response to an oral challenge test with profilin, and after a refinement using a mathematical metabolomic model. PBMCs were used for microarray analysis.We found a set of transcripts and metabolites that were specific for the "severe" phenotype. By metabolomics, a decrease in carbohydrates and pyruvate and an increase in lactate were detected, suggesting aerobic glycolysis. Other metabolites were incremented in "severe" group: lysophospholipids, sphingosine-1-phosphate, sphinganine-1-phosphate, and lauric, myristic, , and oleic fatty acids. On the other hand, carnitines were decreased along severity. Significant transcripts in the "severe" group were found to be downregulated and were associated with platelet functions, protein synthesis, histone modification, and fatty metabolism.We have found evidence that points to the association of severe allergic with platelet functions alteration, together with reduced protein synthesis, and switch of immune cells to aerobic glycolysis.© 2018 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

Keyword: inflammation

The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis.

Hyperglycemia and hyperlipidemia (glycolipotoxicity)-triggered islet β-cell dysfunction is known to drive the progression of obesity-related type 2 diabetes, however the underlying mechanisms have not been clearly elucidated. The current study aimed to investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP-5) in islet cells under glucolipotoxic conditions. Using gene overexpression and knockdown approaches, we demonstrated that MKP-5 could alleviate glucolipotoxicity-induced apoptosis via the endoplasmic reticulum (ER) stress and mitochondrial apoptosis pathways owing to the altered regulation of caspase family members and ER stress-related molecules in MIN6 and primary islet cells. Overexpression of MKP-5 reversed the glucose and (GP)-induced impairment of insulin secretion as well as the abnormal decreases in the expression of islet functional genes, thereby maintaining the normal insulin secretory functionality, whereas the absence of MKP-5 aggravated islet cell dysfunction. In parallel, the production of ROS and increased -associated genes in response to GP were also reduced upon MKP-5 overexpression. Further, inhibition of JNK or P38 MAPK pathways resisted to glucolipotoxicity observed in MKP-5 knockdown MIN6 cells. These findings indicate that MKP-5 is an important mediator for glucolipotoxicity-induced islet cell dysfunction and apoptosis, with JNK and P38 as the critical downstream pathways.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: inflammation

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 diabetes.

Metabolic stress-induced low-grade chronic plays an important role in the development of insulin-resistance and type 2 diabetes (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the obesity-induced metabolic stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: inflammation

Nicotinamide riboside regulates and mitochondrial markers in AML12 hepatocytes.

The NAD precursor nicotinamide riboside (NR) is a type of vitamin B found in cow\'s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer\'s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on and mitochondrial biogenesis in AML12 mouse hepatocytes.A subset of hepatocytes was treated with (PA; 250 µM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 µM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed.Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes.These data demonstrated that NR attenuated hepatic and increased levels of mitochondrial markers in hepatocytes.

Keyword: inflammation

Molecular Targets of Fatty Ethanolamides in Asthma.

Asthma is a common allergic pathology of the respiratory tract that requires the study of mechanisms underlying it, due to severe forms of the disease, which are refractory to therapy. The review is devoted to the search for molecular targets of fatty ethanolamides in asthma, in particular palmitoylethanolamide (PEA), which has been successfully used in the treatment of chronic inflammatory and neurodegenerative diseases, in the pathogenesis of which the nervous and immune systems are involved. Recently, the potentially important role of neuro-immune interactions in the development of allergic reactions has been established. Many of the clinical symptoms accompanying allergic airway are the result of the activation of neurons in the airways, so the attention of researchers is currently focused on neuro-immune interactions, which can play an important role in asthma pathophysiology. A growing number of scientific works confirm that the key molecule in the implementation of these inter-systemic interactions is nerve growth factor (NGF). In addition to its classic role in nervous system physiology, NGF is considered as an important factor associated with the pathogenesis of allergic diseases, particularly asthma, by regulating of mast cell differentiation. In this regard, NGF can be one of the targets of PEA in asthma therapy. PEA has a biological effect on the nervous system, and affects the activation and the degranulation of mast cells.

Keyword: inflammation

Anti-inflammatory activity of Jefea gnaphalioides (a. gray), Astereaceae.

is a symptom associated with many diseases. This symptom is treated with steroidal and non-steroidal anti-inflammatory drugs, which can cause severe side effects when used as long-term treatments. Natural products are an alternative source of new compounds with anti-inflammatory activity. Jefea gnaphalioides (Astereaceae) (A. Gray) is a plant species used to treat inflammatory problems, in Mexico. This study determined the anti-inflammatory activity and the composition of the methanol extract of Jefea gnaphalioides (MEJG).The extract was obtained by heating the plant in methanol at boiling point for 4\u2009h, and then the solvent was evaporated under vacuum (MEJG). The derivatization of the extract was performed using Bis-(trimethylsilyl) trifluoroacetamide, and the composition was determined by GC-MS. Total Phenols and flavonoids were determined by Folin-Ciocalteu AlCl reaction respectively. The antioxidant activity of MEJG was determined by DPPH method. The acute and chronic anti-inflammatory effects were evaluated on a mouse ear edema induced with 12-O-Tetradecanoylphorbol-13-acetate (TPA). Acute oral toxicity was tested in mice at doses of MEJG of 5000, 2500 and 1250\u2009mg/kg. The levels of NO, TNF-α, IL-1β and IL-6 were determinate in J774A.1 macrophages stimulated by Lipopolysaccharide. The production of inflammatory interleukins was measured using commercial kits, and nitric oxide was measured by the Griess reaction.The anti-inflammatory activity of MEJG in acute TPA-induced ear edema was 80.7\u2009±\u20092.8%. This result was similar to the value obtained with indomethacin (IND) at the same dose (74.3\u2009±\u20092.8%). In chronic TPA-induced edema at doses of 200\u2009mg/kg, the inhibition was 45.7%, which was similar to that obtained with IND (47.4%). MEJG have not toxic effects even at a dose of 5000\u2009mg/kg. MEJG at 25, 50, 100 and 200\u2009μg/mL decreased NO, TNF-α, IL-1β and IL-6 production in macrophages stimulated with LPS. The major compounds in MEJG were α-D-Glucopyranose (6.71%), (5.59%), D-(+)-Trehalose (11.91%), Quininic (4.29%) and Aucubin (1.17%). Total phenolic content was 57.01\u2009mg GAE/g and total flavonoid content was 35.26\u2009mg QE/g MEJG had antioxidant activity.MEJG has anti-inflammatory activity.

Keyword: inflammation

The Ethanol Extract of Larvae, Containing Fatty acids and Amino acids, Exerts Anti-Asthmatic Effects through Inhibition of the GATA-3/Th2 Signaling Pathway in Asthmatic Mice.

larvae (HD), a natural product from an insect resource, possesses many pharmacological properties, including anticoagulant, antitumor, anti-inflammatory, and analgesic activity. The major bioactive ingredients include oleic , , palmitoleic , linoleic , proline, and glutamic . Although HD is associated with immunoregulatory activities in allergic diseases, the therapeutic mechanisms of the action of HD in allergic diseases have not been investigated. The aim of this study was to evaluate the anti-asthmatic potential of HD in an ovalbumin (OVA)-induced mouse model of allergic asthma. Moreover, the anti-inflammatory potential of HD was examined to identify a plausible mechanism of action of HD in vitro. HD strongly reduced goblet cell hyperplasia, eosinophil infiltration, and reactive oxygen species (ROS), which reduced airway hyperresponsiveness (AHR), , and the expression of Th2 cytokines (IL-5 and IL-13) in bronchoalveolar lavage fluid (BALF). The expression of IL-5, IL-4, eotaxin-2, lysyl oxidase-like 2 (loxl2), and GATA-binding protein 3 (GATA-3) was attenuated in the lungs. In an in vitro assay, HD exerted immunomodulatory effects through the suppression of Th2 cytokines (IL-5, IL-13), IL-17, and tumor necrosis factor (TNF)-α production through downregulation of GATA-3 expression in EL-4 T cells. These findings suggest that the anti-asthmatic activity of HD may occur through the suppression of Th2 cytokines and total Immunoglobulin E (IgE) production by inhibition of the GATA-3 transcription pathway. Our results suggest that HD may be a potential alternative therapy, or a novel therapeutic traditional medicine, for the treatment of allergic asthma.

Keyword: inflammation

Triterpenoids Improve Diabetes-Induced Hepatic the Rho-Kinase-Dependent Pathway.

This study aimed to assess the effects of triterpene extract of (Batal.) Iljinskaja (CPT) on diabetes-induced hepatic and to unveil the underlying mechanisms. Diabetes in db/db mice was alleviated after CPT administration, as assessed by the oral glucose tolerance test. In addition, treatment with CPT dramatically reduced serum insulin, aspartate amino-transaminase, alanine aminotransferase, triglyceride, and total cholesterol amounts. Besides, serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were also reduced after CPT administration. Western blot analysis revealed that CPT treatment significantly reversed the protein expression levels of Rho, ROCK1, ROCK2, p-P65, p-IκBα, p-IKKα, and p-IKKβ in liver samples obtained from db/db mice. Upon stimulation, the protective effects of CPT on the liver were further assessed in HepG2 and LO2 cells, and no appreciable cytotoxic effects were found. Therefore, these findings indicate that CPT alleviates liver Rho-kinase signaling. Metformin (PubChem CID: 4091); Fasudil (PubChem CID: 3547); (PubChem CID: 985).

Keyword: inflammation

Early and reversible changes to the hippocampal proteome in mice on a high-fat diet.

The rise in global obesity makes it crucial to understand how diet drives obesity-related health conditions, such as premature cognitive decline and Alzheimer\'s disease (AD). In AD hippocampal-dependent episodic memory is one of the first types of memory to be impaired. Previous studies have shown that in mice fed a high-fat diet (HFD) episodic memory is rapidly but reversibly impaired.In this study we use hippocampal proteomics to investigate the effects of HFD in the hippocampus. Mice were fed either a low-fat diet (LFD) or HFD containing either 10% or 60% (Kcal) from fat for 3\u2009days, 1\u2009week or 2\u2009weeks. One group of mice were fed the HFD for 1\u2009week and then returned to the LFD for a further week. Primary hippocampal cultures were challenged with (PA), the most common long-chain saturated FA in the Western diet, and with the anti-inflammatory, n-3 polyunsaturated FA, docosahexaenoic (DHA), or a combination of the two to ascertain effects of these fatty acids on dendritic structure.HFD-induced changes occur in hippocampal proteins involved in metabolism, , cell stress, cell signalling, and the cytoskeleton after 3\u2009days, 1\u2009week and 2\u2009weeks of HFD. Replacement of the HFD after 1\u2009week by a low-fat diet (LFD) for a further week resulted in partial recovery of the hippocampal proteome. Microtubule-associated protein 2 (MAP2), one of the earliest proteins changed, was used to investigate the impact of fatty acids (FAs) on hippocampal neuronal morphology. PA challenge resulted in shorter and less arborised dendrites while DHA had no effect when applied alone but counteracted the effects of PA when FAs were used in combination. Dendritic morphology recovered when PA was removed from the cell culture media.This study provides evidence for the rapid and reversible effects of diet on the hippocampal proteome and the impact of PA and DHA on dendritic structure.

Keyword: inflammation

MiR27a Promotes the Development of Macrophage-like Characteristics in 3T3-L1 Preadipocytes.

Recruitment and polarization of classically activated (M1) macrophages within adipose tissue contribute to chronic low-grade in obesity. Adipose tissue precursor cells exhibit the capacity to develop macrophage-like characteristics and adipocyte-derived miR27a is known to promote reprogramming of somatic cells. It was unknown whether exogenous addition of miR27a promote the development of macrophage-like characteristics of adipose precursor cells. We examined macrophage surface antigen, phagocytosis and migration ability in 3T3-L1 preadipocytes transfected with miR27a mimics. Transfection of 3T3-L1 preadipocytes with miR27a mimics increased phagocytosis and migration and increased the number of cells expressing the macrophage makers F4/80 and MHC compared to controls. M2 and CD206 macrophage markers were unaltered. In addition, transfection of 3T3-L1 preadipocytes with miR27a mimics reduced PPARγ expression, activated NF-κB and promoted secretion of the inflammatory cytokines MCP-1, TNF-α and IL-1β compared to controls. The level of anti-inflammatory factors Arg-1, IL-10, Ym1 and Fizz1 were unaltered. Secretion of miR27a was increased in conditioned medium prepared from -treated differentiated 3T3-L1 adipocytes compared to controls. Incubation of 3T3-L1 preadipocytes with this conditioned medium increased phagocytosis and migration compared to controls. Finally, conditioned medium prepared from differentiated 3T3-L1 adipocytes transfection with miR27a inhibitors reduced phagocytosis and migration in 3T3-L1 preadipocytes compared to controls. The data indicate that PPARγ agonists may reverse the activation of NF-κB pathway mediated by miR27a overexpression and reduce phagocytosis and migration of adipose precursor cells. In addition, miR27a may promote the development of macrophage-like characteristics in 3T3-L1 preadipocytes.

Keyword: inflammation

Analysis of Fatty Esters of Hydroxyl Fatty in Selected Plant Food.

Metabolic syndrome, characterized by obesity, low-grade , insulin resistance, hyperglycemia, dyslipidemia and hypertension, is a major risk factor for cardiovascular mortality. Preclinical studies on recently discovered classes of lipids - fatty esters of hydroxy fatty acids (FAHFA) have revealed their anti-inflammatory and insulin-sensitizing potential. The FAHFA levels are significantly decreased in insulin-resistant individuals, their application exhibited anti-inflammatory effects and restoring the glucose-insulin homeostasis. The aim of our research was to analyze the overall FAHFA composition in a common diet, as only a partial FAHFA composition has been revealed so far (only the PAHSA subclass was analyzed in a few foods). A new approach to the FAHFAs analysis includes nano-LC and post-column modifier followed by negative ion mass spectrometry, in order to obtain maximum sensitivity. Analysis of different foods - oat (whole grain, coarse flakes and fine flakes), apple, clementine, lemon, strawberry, blueberry, mango, kiwi, avocado, pineapple, banana, onion, garlic, cherry tomato, carrot, parsley root, pepper and radish - exhibited wide inter-food variation in the FAHFA profiles. Sixteen analyzed FAHFAs (, oleic, palmitoleic and stearic hydroxy-esters) showed microgram to low nanogram levels (0.165\xa0ng/g - 32\xa0μg/g FW), with the highest abundancy in oat, clementine, garlic and pineapple. Stearic hydroxy stearic (SAHSA) was the most abundant FAHFA, especially in the food with antioxidative, anti-inflammatory and beneficial metabolic effects. In contrary, the PAHSA - previously proven to have the strongest antihyperglycemic and insulin-sensitizing effects, was not present in some foods (radish, avocado, mango, lemon, cherry tomato, kiwi). Our study proves the importance of overall FAHFA analysis in food (especially in a functional food), because of their potential metabolic benefits and possible future incorporation in special diets.

Keyword: inflammation

Histone demethylase UTX is a therapeutic target for diabetic kidney disease.

Diabetic kidney disease (DKD) is a major complication of diabetes. We found that UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase, was upregulated in the renal mesangial and tubular cells of diabetic mice and DKD patients. In cultured renal mesangial and tubular cells, UTX overexpression promoted -induced elevation of and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. We found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes and apoptosis; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the diabetes-induced renal abnormalities in db/db mice, an animal model of type 2 diabetes. These results revealed the possible mechanisms underlying the regulation of histone methylation in DKD and suggest UTX as a potential therapeutic target for DKD.Diabetic kidney disease (DKD) is a microvascular complication of diabetes and the leading cause of end-stage kidney disease worldwide without effective therapy available. UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase that removes the di- and tri-methyl groups from histone H3K27, plays important biological roles in gene activation, cell fate control and life span regulation in Caenorhabditis elegans. In the present study, we report upregulated UTX in the kidneys of diabetic mice and DKD patients. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the diabetes-induced renal dysfunction, abnormal morphology, , apoptosis and DNA damage in db/db mice, comprising an animal model of type 2 diabetes. In cultured renal mesanglial and tubular cells, UTX overexpression promoted induced elevation of and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. Mechanistically, we found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Collectively, our results suggest UTX as a potential therapeutic target for DKD.© 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

Keyword: inflammation

Fatty activates NLRP3 inflammasomes in mouse Kupffer cells through mitochondrial DNA release.

Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease in many developed and developing countries worldwide. It has been well established that the chronic sterile caused by the NLRP3 inflammasome is closely related to NAFLD development. Kupffer cells (KCs) are involved in the pathogenesis of various liver diseases. We used methionine choline-deficient diets to establish a mouse nonalcoholic steatohepatitis (NASH) model. The expression and formation of the NLRP3 inflammasome in the KCs from the mouse and cell models were determined by Western blotting and co-immunoprecipitation. Evidence of mitochondrial DNA (mtDNA) release was determined by live cell labeling and imaging. KCs and the NLRP3 inflammasome exerted proinflammatory effects on the development and progression of NASH through secretion of the proinflammatory cytokine IL-1β. NLRP3, ASC and Caspase-1 protein expression levels in KCs from NASH mouse livers were significantly higher than those in KCs from NLRP3 mice, and the number of NLRP3 inflammasome protein complexes was significantly higher in KCs from NASH mouse livers, whereas these protein complexes could not be formed in NLRP3 mice. In in vitro experiments, (PA) decreased the mitochondrial membrane potential and subsequently induced mtDNA release from the mitochondria to the cytoplasm. NLRP3 inflammasome expression was substantially increased, and mtDNA-NLRP3 inflammasome complexes formed upon PA stimulation. Our data suggest that mtDNA released from mitochondria during PA stimulation causes NLRP3 inflammasome activation, providing a missing link between NLRP3 inflammasome activation and NASH development, via binding of cytosolic mtDNA to the NLRP3 inflammasome.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

The regulation of -related genes after and DHA treatments is not mediated by DNA methylation.

Fatty acids (FAs) are known to participate in body inflammatory responses. In particular, saturated FAs such as (PA) induce inflammatory signals in macrophages, whereas polyunsaturated FAs, including docosahexaenoic (DHA), have been related to anti-inflammatory effects. Several studies have suggested a role of fatty acids on DNA methylation, epigenetically regulating gene expression in processes. Therefore, this study investigated the effect of PA and DHA on the -related genes on human macrophages. In addition, a second aim was to study the epigenetic mechanism underlying the effect of FAs on the inflammatory response. For these purposes, human acute monocytic leukaemia cells (THP-1) were differentiated into macrophages with 12-O-tetradecanoylphorbol-13-acetate (TPA), followed by an incubation with PA or DHA. At the end of the experiment, mRNA expression, protein secretion, and CpG methylation of the following inflammatory genes were analysed: interleukin 1 beta (IL1B), tumour necrosis factor (TNF), plasminogen activator inhibitor-1 (SERPINE1) and interleukin 18 (IL18). The results showed that the treatment with PA increased IL-18 and TNF-α production. Contrariwise, the supplementation with DHA reduced IL-18, TNF-α and PAI-1 secretion by macrophages. However, the incubation with these fatty acids did not apparently modify the DNA methylation status of the investigated genes in the screened CpG sites. This research reveals that PA induces important pro-inflammatory markers in human macrophages, whereas DHA decreases the inflammatory response. Apparently, DNA methylation is not directly involved in the fatty -mediated regulation of the expression of these -related genes.

Keyword: inflammation

9-PAHSA promotes browning of white fat via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract obesity in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white fat. 9-PAHSA, an endogenous mammalian lipid, which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown fat specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown fat markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of obesity.Copyright © 2018. Published by Elsevier Inc.

Keyword: inflammation

Linoleic rescues microglia triggered by saturated fatty .

Elevated saturated free fatty levels during over-nutrition lead to hypothalamic , which perturbs energy homeostasis. Whether brain-derived metabolites are coupled to the development of obesity pathogenesis during the early over-nutrition period has not been thoroughly investigated. In this study, we found increased linoleic , an unsaturated fatty , in both the whole brain and hypothalamus of mice fed a high-fat diet for 4 weeks. Furthermore, we observed that linoleic effectively reversed the inflammatory responses induced by treatment in microglial cells. Collectively, this study suggests the reversible function of linoleic on brain in association with microglial activation during short-term exposure to a high-fat diet.Copyright © 2019. Published by Elsevier Inc.

Keyword: inflammation

Hsp72 protects against liver injury via attenuation of hepatocellular death, oxidative stress, and JNK signaling.

Heat shock protein (Hsp) 72 is a molecular chaperone that has broad cytoprotective functions and is upregulated in response to stress. To determine its hepatic functions, we studied its expression in human liver disorders and its biological significance in newly generated transgenic animals.Double transgenic mice overexpressing Hsp72 (gene Hspa1a) under the control of a tissue-specific tetracycline-inducible system (Hsp72-LAP mice) were produced. Acute liver injury was induced by a single injection of acetaminophen (APAP). Feeding with either a methionine choline-deficient (MCD; 8\u202fweeks) or a 3,5-diethoxycarbonyl-1,4-dihydrocollidine-supplemented diet (DDC; 12\u202fweeks) was used to induce lipotoxic injury and Mallory-Denk body (MDB) formation, respectively. Primary hepatocytes were treated with .Patients with non-alcoholic steatohepatitis and chronic hepatitis C infection displayed elevated HSP72 levels. These levels increased with the extent of hepatic and HSP72 expression was induced after treatment with either interleukin (IL)-1β or IL-6. Hsp72-LAP mice exhibited robust, hepatocyte-specific Hsp72 overexpression. Primary hepatocytes from these animals were more resistant to isolation-induced stress and Hsp72-LAP mice displayed lower levels of hepatic injury in vivo. Mice overexpressing Hsp72 had fewer APAP protein adducts and were protected from oxidative stress and APAP-/MCD-induced cell death. Hsp72-LAP mice and/or hepatocytes displayed significantly attenuated Jnk activation. Overexpression of Hsp72 did not affect steatosis or the extent of MDB formation.Our results demonstrate that HSP72 induction occurs in human liver disease, thus, HSP72 represents an attractive therapeutic target owing to its broad hepatoprotective functions.HSP72 constitutes a stress-inducible, protective protein. Our data demonstrate that it is upregulated in patients with chronic hepatitis C and non-alcoholic steatohepatitis. Moreover, Hsp72-overexpressing mice are protected from various forms of liver stress.Copyright © 2018 European Association for the Study of the Liver. All rights reserved.

Keyword: inflammation

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated .

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and . 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated .

Keyword: inflammation

Adelmidrol + sodium hyaluronate in IC/BPS or conditions associated to chronic urothelial . A translational study.

Interstitial cystitis/painful bladder syndrome (IC/PBS) is a chronic bladder condition characterized by frequent urination, bladder and pain. It is a particular challenging disease and a clear unmet medical need in terms of identifying new therapeutic strategies. The aim of study was to evaluate the anti-inflammatory effects of intravesical Vessilen (a new formulation of 2% adelmidrol (the diethanolamide derivative of azelaic ) + 0.1% sodium hyaluronate) administration in rodent models of IC/BPS and in IC/BPS patients or other bladder disorders. Acute and chronic animal models of cystitis were induced by a single or repetitive intraperitoneal injections of cyclophosphamide (CYP); patients with IC/BPS or with bladder pain syndrome associated with symptoms of the lower urinary tract treated once weekly by bladder instillation of Vessilen for 8 weeks. CYP instillation caused macroscopic and histological bladder alterations, inflammatory infiltrates, increased mast cell numbers, bladder pain, increased expression of nitrotyrosine, decreased expression of endothelial tight junction zonula occludens-1. Intravesical Vessilen® treatment was able to ameliorate CYP induced bladder and pain by inhibiting nuclear factor-κB pathway and inflammatory mediator levels as well as reduced mechanical allodynia and nerve growth factor levels. A significant improvement in quality of life and symptom intensity were evident in patients with IC/BPS or other bladder disorders treated with Vessilen. Vessilen could be a new therapeutic approach for human cystitis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammation

The phytochemical polydatin ameliorates non-alcoholic steatohepatitis by restoring lysosomal function and autophagic flux.

Impaired autophagic degradation of intracellular lipids is causally linked to the development of non-alcoholic steatohepatitis (NASH). Pharmacological agents that can restore hepatic autophagic flux could therefore have therapeutic potentials for this increasingly prevalent disease. Herein, we investigated the effects of polydatin, a natural precursor of resveratrol, in a murine nutritional model of NASH and a cell line model of steatosis. Results showed that oral administration of polydatin protected against hepatic lipid accumulation and alleviated and hepatocyte damage in db/db mice fed methionine-choline deficient diet. Polydatin also alleviated -induced lipid accumulation in cultured hepatocytes. In both models, polydatin restored lysosomal function and autophagic flux that were impaired by NASH or steatosis. Mechanistically, polydatin inhibited mTOR signalling and up-regulated the expression and activity of TFEB, a known master regulator of lysosomal function. In conclusion, polydatin ameliorated NASH through restoring autophagic flux. The polydatin-regulated autophagy was associated with inhibition of mTOR pathway and restoration of lysosomal function by TFEB. Our study provided affirmative preclinical evidence to inform future clinical trials for examining the potential anti-NASH effect of polydatin in humans.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: inflammation

Downregulation of MALAT1 alleviates saturated fatty -induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis.

The increased level of saturated fatty acids (SFAs) is found in patients with diabetes, obesity, and other metabolic disorders. SFAs can induce lipotoxic damage to cardiomyocytes, but the mechanism is unclear. The long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acts as a key regulator in (PA)-induced hepatic steatosis, but its role in PA-induced myocardial lipotoxic injury is still unknown. The aim of this study was to explore the role and underlying mechanism of MALAT1 in PA-induced myocardial lipotoxic injury. MALAT1 expression in PA-treated human cardiomyocytes (AC16 cells) was detected by RT-qPCR. The effect of MALAT1 on PA-induced myocardial injury was measured by Cell Counting Kit-8, lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) assays. Apoptosis was detected by flow cytometry. The activities of cytokines and nuclear factor (NF)-κB were detected by enzyme-linked immunosorbent assay. The interaction between MALAT1 and miR-26a was evaluated by a luciferase reporter assay and RT-qPCR. The regulatory effects of MALAT1 on high mobility group box 1 (HMGB1) expression were evaluated by RT-qPCR and western blotting. MALAT1 was significantly upregulated in cardiomyocytes after PA treatment. Knockdown of MALAT1 increased the viability of PA-treated cardiomyocytes, decreased apoptosis, and reduced the levels of LDH, CK-MB, TNF-α, and IL-1β. Moreover, we found that MALAT1 specifically binds to miR-26a and observed a reciprocal negative regulatory relationship between these factors. We further found that the downregulation of MALAT1 represses HMGB1 expression, thereby inhibiting the activation of the Toll-like receptor 4 (TLR4)/NF-κB-mediated inflammatory response. These repressive effects were rescued by an miR-26a inhibitor. We demonstrate that MALAT1 is induced by SFAs and its downregulation alleviates SFA-induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis. Our findings provide new insight into the mechanism underlying myocardial lipotoxic injury.

Keyword: inflammation

High-density lipoprotein ameliorates -induced lipotoxicity and oxidative dysfunction in H9c2 cardiomyoblast cells via ROS suppression.

High levels circulating saturated fatty acids are associated with diabetes, obesity and hyperlipidemia. In heart, the accumulation of saturated fatty acids has been determined to play a role in the development of heart failure and diabetic cardiomyopathy. High-density lipoprotein (HDL) has been reported to possess key atheroprotective biological properties, including cellular cholesterol efflux capacity, anti-oxidative and anti-inflammatory activities. However, the underlying mechanisms are still largely unknown. Therefore, the aim of the present study is to test whether HDL could protect (PA)-induced cardiomyocyte injury and explore the possible mechanisms.H9c2 cells were pretreated with HDL (50-100\u2009μg/ml) for 2\u2009h followed by PA (0.5\u2009mM) for indicated time period. Our results showed that HDL inhibited PA-induced cell death in a dose-dependent manner. Moreover, HDL rescued PA-induced ROS generation and the phosphorylation of JNK which in turn activated NF-κB-mediated inflammatory proteins expressions. We also found that PA impaired the balance of BCL family proteins, destabilized mitochondrial membrane potential, and triggered subsequent cytochrome c release into the cytosol and activation of caspase 3. These detrimental effects were ameliorated by HDL treatment.PA-induced ROS accumulation and results in cardiomyocyte apoptosis and . However, HDL attenuated PA-induced lipotoxicity and oxidative dysfunction via ROS suppression. These results may provide insight into a possible molecular mechanism underlying HDL suppression of the free fatty -induced cardiomyocyte apoptosis.

Keyword: inflammation

Role of the saturated fatty palmitate in the interconnected hypothalamic control of energy homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body energy and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, obesity, and type 2 diabetes mellitus. Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and energy homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal ; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: inflammation

Anthocyanins ameliorate palmitate-induced and insulin resistance in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade leading to insulin resistance. Adipocyte differentiation inhibitors are expected to be effective in preventing obesity and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from metabolic stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, , and insulin resistance in 3T3-L1 adipocytes. ACN extract pretreatment reduces lipid accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces insulin resistance by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce insulin pathway with levels higher than insulin control cells, supporting an insulin sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against obesity comorbidities, due to their protective effects against /insulin resistance in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and insulin resistance conditions related to obesity.© 2019 John Wiley & Sons, Ltd.

Keyword: inflammation

1,25-Dihydroxyvitamin D regulates lipid metabolism and glucose utilization in differentiated 3T3-L1 adipocytes.

It is well established that adipose tissue can both store and metabolize vitamin D. The active form of vitamin D, 1,25 dihydroxyvitamin D [1,25(OH)D], regulates adipocyte differentiation and , highlighting the multifaceted role that vitamin D plays in adipose tissue physiology. However, there is limited evidence regarding vitamin D regulation of mature adipocyte lipid metabolism. We hypothesize that 1,25(OH)D alters lipid and glucose metabolism in differentiated 3T3-L1 adipocytes to reduce triacylglycerol (TAG) accumulation. In this study, 1,25(OH)D (10\u202fnmol/L) stimulated a 21% reduction in TAG accumulation in differentiated 3T3-L1 adipocytes after 4\u202fdays (P\u202f=\u202f.01) despite a significant increase in fatty uptake (P\u202f<\u202f.01). Additionally, 1,25(OH)D stimulated a 2.5-fold increase in CO production from [1-C] (P\u202f<\u202f.01), indicative of an elevated rate of fatty β-oxidation, while stimulating a 9% reduction in de novo fatty synthesis (P\u202f=\u202f.03). Interestingly, d-[U-C]glucose incorporation into fatty acids was reduced by 30% in response to 1,25(OH)D (P\u202f<\u202f.01), indicating a reduced contribution of glucose as a substrate for de novo lipogenesis. Consistent with these findings, mRNA expression of the anaplerotic enzyme pyruvate carboxylase was reduced by 41% (P\u202f<\u202f.01). In summary, 1,25(OH)D stimulated fatty oxidation and reduced TAG accumulation in differentiated adipocytes. Furthermore, 1,25(OH)D reduced glucose utilization as a substrate for fatty synthesis potentially by downregulating pyruvate carboxylase and stimulating glucose disposal as glycerol. Collectively, these 1,25(OH)D-induced changes in lipid metabolism and glucose utilization may contribute to the reduction in TAG accumulation and be protective against excessive fat mass accumulation and associated metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Palmitate Induces an Anti-Inflammatory Response in Immortalized Microglial BV-2 and IMG Cell Lines that Decreases TNFα Levels in mHypoE-46 Hypothalamic Neurons in Co-Culture.

Elevated levels of saturated fatty acids (SFA) induce a state of neuroinflammation in the hypothalamus. It has been suggested that microglia sense palmitate, a prevalent circulating SFA, and act as mediators of this inflammatory process by communicating with neurons, particularly those involved in appetite regulation. In this study, we examined the inflammatory response to palmitate in immortalized microglial cell lines, BV-2 and IMG, and the subsequent effects on inflammatory gene expression in a model of NPY/AgRP neurons, mHypoE-46.The BV-2 cells were treated with 50 µM palmitate for 4 and 24 h, and the transcriptional regulation of markers for and cellular stress was assessed using an RT2 Profiler PCR Array. Select genes were verified with qRT-PCR. The BV-2 and IMG cells were then co-cultured using 1.0-µm cell culture inserts with an immortalized hypothalamic cell line, mHypoE-46, to investigate potential intercellular communication between microglia and neurons.We found that palmitate increased the mRNA levels of specific inflammatory genes, and a general anti-inflammatory profile was revealed in the microglia cells. The mRNA changes in TNFα at 4 and 24 h in BV-2 cells were abrogated with the toll-like receptor 4 (TLR4) inhibitor, TAK-242, indicating the involvement of TLR4. Co-culture of mHypoE-46 neurons with microglia pre-treated with palmitate resulted in repression of TNFα expression in the hypothalamic neurons. As palmitate significantly increased IL-13 expression in microglia, the effect of this cytokine was tested in mHypoE-46 neurons. The addition of IL-13 to neuronal cultures normalized the palmitate-mediated increase in IL-6 and AgRP expression, suggesting that microglia may protect surrounding neurons, at least in part, through the release of IL-13.These results suggest a potential anti-inflammatory role of microglia towards the palmitate-induced neuroinflammation, and potentially energy homeostasis, in hypothalamic neurons.© 2018 ©2018S. Karger AG, Basel.

Keyword: inflammation

Cangju Qinggan Jiangzhi Decoction Reduces the Development of NonAlcoholic Steatohepatitis and Activation of Kupffer Cells.

Nonalcoholic steatohepatitis (NASH) is defined as lipid accumulation with hepatic injury, and early to moderate fibrosis. Kupffer cells play a crucial role in promoting hepatic , which further facilitates the development of NASH. Here we investigated the effects of Cangju Qinggan Jiangzhi decoction (CQJD) on high fat diet (HFD) and methionine-choline deficient (MCD) induced mouse NASH pathogenesis.Mouse NASH models were developed by HFD and MCD diet. The treated mice were divided into three groups: the control group (n = 10), the low-dose CQJD treatment group (n = 10) and the high-dose CQJD treatment group (n = 10). The hepatic injury, , and apoptotic molecules were evaluated by H&E staining, immunohistochemistry and real-time PCR. Kupffer cells were isolated from control mice and CQJD-treated mice after stimulation by lipopolysaccharide (LPS) and/or . The level of the inflammatory cytokines TNFα, IL1β, and CCL2 was measured by ELISA.The HFD-fed mice displayed significant metabolic, inflammatory, and oxidative stress-related alterations due to hepatic lipid accumulation. CQJD treatment largely normalized the hepatic injury, lowered the ALT/AST level, and reduced the severity of liver , as revealed by the decreased inflammatory cytokines levels. In vitro, CQJD blocked the activation of LPS- or -primed Kupffer cells in a dose-dependent manner. In the MCD diet-induced NASH mice, similar therapeutic effects of CQJD were also observed.CQJD ameliorates mouse nonalcoholic steatohepatitis. The reduction in liver injury and induced by CQJD is associated with reduced activation of Kupffer cells. Our results suggest that CQJD is a promising therapeutic strategy in clinical steatohepatitis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: inflammation

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet .

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet , and that GPR120 activation may serve as a preventative and therapeutic target for obesity and diabetes.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: inflammation

Fatty metabolism in the host and commensal bacteria for the control of intestinal immune responses and diseases.

Intestinal tissue has a specialized immune system that exhibits an exquisite balance between active and suppressive responses important for the maintenance of health. Intestinal immunity is functionally affected by both diet and gut commensal bacteria. Here, we review the effects of fatty acids on the regulation of intestinal immunity and immunological diseases, revealing that dietary fatty acids and their metabolites play an important role in the regulation of allergy, , and immunosurveillance in the intestine. Several lines of evidence have revealed that some dietary fatty acids are converted to biologically active metabolites by enzymes not only in the host but also in the commensal bacteria. Thus, biological interaction between diet and commensal bacteria could form the basis of a new era in the control of host immunity and its associated diseases.

Keyword: inflammation

Celastrol reverses (PA)-caused TLR4-MD2 activation-dependent insulin resistance via disrupting MD2-related cellular binding to PA.

Elevated plasma statured fatty acids (FFAs) cause TLR4/MD2 activation-dependent and insulin tolerance, which account for the occurrence and development of obesity. It has been confirmed that statured (PA) (the most abundant FFA) could bind MD2 to cause cellular . The natural compound celastrol could improve obesity, which is suggested via inhibiting , yet the detailed mechanism for celastrol is still unclear. As celastrol is reported to directly target MD2, we thought disrupting the binding between FFAs and MD2 might be one of the ways for celastrol to inhibit FFAs-caused and insulin resistance. In this study, we found evidence to support our hypothesis: celastrol could reverse PA-caused TLR4/MD2 activation-dependent insulin resistance, as determined by glucose-lowering ability, cellular glucose uptake, insulin action-related proteins and TLR4/MD2/NF-κB activation. Bioinformatics and cellular experiments showed that both celastrol and PA could bind MD2, and that celastrol could expel PA from cells. Finally, celastrol could reverse high fat diet caused hyperglycemia and obesity, and liver NF-kB activations. Taking together, we proved that celastrol could reverses PA-caused TLR4-MD2 activation-dependent insulin resistance via disrupting PA binding to MD2.© 2018 Wiley Periodicals, Inc.

Keyword: inflammation

Frontline Science: Specialized proresolving lipid mediators inhibit the priming and activation of the macrophage NLRP3 inflammasome.

The prototypic proinflammatory cytokine IL-1β plays a central role in innate immunity and inflammatory disorders. The formation of mature IL-1β from an inactive pro-IL-1β precursor is produced via nonconventional multiprotein complexes called the inflammasomes, of which the most common is the nucleotide-binding domain leucine-rich repeat-containing protein 3 (NLRP3) inflammasome composed by NLRP3, (ASC) apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (CARD), and caspase-1. Specialized proresolving mediators (SPMs) promote resolution of , which is an essential process to maintain host health. SPMs prevent excessive by terminating the inflammatory response and returning to tissue homeostasis without immunosupression. This study tested the hypothesis that modulation of the NLRP3 inflammasome in macrophages is one mechanism involved in the SPM-regulated processes during resolution. Our findings demonstrate that the SPM resolvin D2 (RvD2) suppressed the expression of pro-IL-1β and reduced the secretion of mature IL-1β in bone marrow-derived macrophages challenged with LPS+ATP (classical NLRP3 inflammasome model) or LPS+palmitate (lipotoxic model). Similar findings were observed in thioglycolate-elicited peritoneal macrophages, in which RvD2 remarkably reduced ASC oligomerization, inflammasome assembly, and caspase-1 activity. In vivo, in a self-resolving zymosan A-induced peritonitis model, RvD2 blocked the NLRP3 inflammasome leading to reduced release of IL-1β into the exudates, repression of osteopontin, and MCP-1 expression and induction of M2 markers of resolution (i.e., CD206 and arginase-1) in peritoneal macrophages. RvD2 inhibitory actions were receptor mediated and were abrogated by a selective GPR18 antagonist. Together, these findings support the hypothesis that SPMs have the ability to inhibit the priming and to expedite the deactivation of the NLRP3 inflammasome in macrophages during the resolution process.©2018 Society for Leukocyte Biology.

Keyword: inflammation

An unexpected link between fatty synthase and cholesterol synthesis in proinflammatory macrophage activation.

Different immune activation states require distinct metabolic features and activities in immune cells. For instance, inhibition of fatty synthase (FASN), which catalyzes the synthesis of long-chain fatty acids, prevents the proinflammatory response in macrophages; however, the precise role of this enzyme in this response remains poorly defined. Consistent with previous studies, we found here that FASN is essential for lipopolysaccharide-induced, Toll-like receptor (TLR)-mediated macrophage activation. Interestingly, only agents that block FASN upstream of acetoacetyl-CoA synthesis, including the well-characterized FASN inhibitor C75, inhibited TLR4 signaling, while those acting downstream had no effect. We found that acetoacetyl-CoA could overcome C75\'s inhibitory effect, whereas other FASN metabolites, including palmitate, did not prevent C75-mediated inhibition. This suggested an unexpected role for acetoacetyl-CoA in that is independent of its role in palmitate synthesis. Our evidence further suggested that acetoacetyl-CoA arising from FASN activity promotes cholesterol production, indicating a surprising link between fatty synthesis and cholesterol synthesis. We further demonstrate that this process is required for TLR4 to enter lipid rafts and facilitate TLR4 signaling. In conclusion, we have uncovered an unexpected link between FASN and cholesterol synthesis that appears to be required for TLR signal transduction and proinflammatory macrophage activation.© 2018 Carroll et al.

Keyword: inflammation

The autophagic degradation of Cav-1 contributes to PA-induced apoptosis and of astrocytes.

The accumulation of (PA), implicated in obesity, can induce apoptotic cell death and of astrocytes. Caveolin-1 (Cav-1), an essential protein for astrocytes survival, can be degraded by autophagy, which is a double-edge sword that can either promote cell survival or cell death. The aim of this study was to delineate whether the autophagic degradation of Cav-1 is involved in PA-induced apoptosis and in hippocampal astrocytes. In this study we found that: (1) PA caused apoptotic death and by autophagic induction; (2) Cav-1 was degraded by PA-induced autophagy and PA induced autophagy in a Cav-1-independent manner; (3) the degradation of Cav-1 was responsible for PA-induced autophagy-dependent apoptotic cell death and ; (4) chronic high-fat diet (HFD) induced Cav-1 degradation, apoptosis, autophagy, and in the hippocampal astrocytes of rats. Our results suggest that the autophagic degradation of Cav-1 contributes to PA-induced apoptosis and of astrocytes. Therefore, Cav-1 may be a potential therapeutic target for central nervous system injuries caused by PA accumulation.

Keyword: inflammation

Prophylactic Palmitoylethanolamide Prolongs Survival and Decreases Detrimental in Aged Mice With Bacterial Meningitis.

Easy-to-achieve interventions to promote healthy longevity are desired to diminish the incidence and severity of infections, as well as associated disability upon recovery. The dietary supplement palmitoylethanolamide (PEA) exerts anti-inflammatory and neuroprotective properties. Here, we investigated the effect of prophylactic PEA on the early immune response, clinical course, and survival of old mice after intracerebral K1 infection. Nineteen-month-old wild type mice were treated intraperitoneally with two doses of either 0.1 mg PEA/kg in 250 μl vehicle solution ( = 19) or with 250 μl vehicle solution only as controls ( = 19), 12 h and 30 min prior to intracerebral K1 infection. The intraperitoneal route was chosen to reduce distress in mice and to ensure exact dosing. Survival time, bacterial loads in cerebellum, blood, spleen, liver, and microglia counts and activation scores in the brain were evaluated. We measured the levels of IL-1β, IL-6, MIP-1α, and CXCL1 in cerebellum and spleen, as well as of bioactive lipids in serum in PEA- and vehicle-treated animals 24 h after infection. In the absence of antibiotic therapy, the median survival time of PEA-pre-treated infected mice was prolonged by 18 h compared to mice of the vehicle-pre-treated infected group ( = 0.031). PEA prophylaxis delayed the onset of clinical symptoms ( = 0.037). This protective effect was associated with lower bacterial loads in the spleen, liver, and blood compared to those of vehicle-injected animals ( ≤ 0.037). PEA-pre-treated animals showed diminished levels of pro-inflammatory cytokines and chemokines in spleen 24 h after infection, as well as reduced serum concentrations of arachidonic and of one of its metabolites, 20-hydroxyeicosatetraenoic . In the brain, prophylactic PEA tended to reduce bacterial titers and attenuated microglial activation in aged infected animals ( = 0.042). Our findings suggest that prophylactic PEA can counteract infection associated detrimental responses in old animals. Accordingly, PEA treatment slowed the onset of infection symptoms and prolonged the survival of old infected mice. In a clinical setting, prophylactic administration of PEA might extend the potential therapeutic window where antibiotic therapy can be initiated to rescue elderly patients.

Keyword: inflammation

Docosahexaenoic antagonizes the boosting effect of on LPS inflammatory signaling by inhibiting gene transcription and ceramide synthesis.

It is well known that saturated fatty acids (SFAs) and unsaturated fatty , in particular omega-3 polyunsaturated fatty acids (n-3 PUFAs), have different effects on inflammatory signaling: SFAs are pro-inflammatory but n-3 PUFAs have strong anti-inflammatory properties. We have reported that (PA), a saturated fatty , robustly amplifies lipopolysaccharide (LPS) signaling to upregulate proinflammatory gene expression in macrophages. We also reported that the increased production of ceramide (CER) via sphingomyelin (SM) hydrolysis and CER de novo synthesis plays a key role in the synergistic effect of LPS and PA on proinflammatory gene expression. However, it remains unclear if n-3 PUFAs are capable of antagonizing the synergistic effect of LPS and PA on gene expression and CER production. In this study, we employed the above macrophage culture system and lipidomical analysis to assess the effect of n-3 PUFAs on proinflammatory gene expression and CER production stimulated by LPS and PA. Results showed that DHA strongly inhibited the synergistic effect of LPS and PA on proinflammatory gene expression by targeting nuclear factor kappa B (NFκB)-dependent gene transcription. Results also showed that DHA inhibited the cooperative effect of LPS and PA on CER production by targeting CER de novo synthesis, but not SM hydrolysis. Furthermore, results showed that myriocin, a specific inhibitor of serine palmitoyltransferase, strongly inhibited both LPS-PA-stimulated CER synthesis and proinflammatory gene expression, indicating that CER synthesis is associated with proinflammatory gene expression and that inhibition of CER synthesis contributes to DHA-inhibited proinflammatory gene expression. Taken together, this study demonstrates that DHA antagonizes the boosting effect of PA on LPS signaling on proinflammatory gene expression by targeting both NFκB-dependent transcription and CER de novo synthesis in macrophages.

Keyword: inflammation

Targeted lipidomics analysis identified altered serum lipid profiles in patients with polymyositis and dermatomyositis.

Polymyositis (PM) and dermatomyositis (DM) are severe chronic autoimmune diseases, characterized by muscle fatigue and low muscle endurance. Conventional treatment includes high doses of glucocorticoids and immunosuppressive drugs; however, few patients recover full muscle function. One explanation of the persistent muscle weakness could be altered lipid metabolism in PM/DM muscle tissue as we previously reported. Using a targeted lipidomic approach we aimed to characterize serum lipid profiles in patients with PM/DM compared to healthy individuals (HI) in a cross-sectional study. Also, in the longitudinal study we compared serum lipid profiles in patients newly diagnosed with PM/DM before and after immunosuppressive treatment.Lipidomic profiles were analyzed in serum samples from 13 patients with PM/DM, 12 HI and 8 patients newly diagnosed with PM/DM before and after conventional immunosuppressive treatment using liquid chromatography tandem mass spectrometry (LC-MS/MS) and a gas-chromatography flame ionization detector (GC-FID). Functional Index (FI), as a test of muscle performance and serum levels of creatine kinase (s-CK) as a proxy for disease activity were analyzed.The fatty (FA) composition of total serum lipids was altered in patients with PM/DM compared to HI; the levels of (16:0) were significantly higher while the levels of arachidonic (20:4, n-6) were significantly lower in patients with PM/DM. The profiles of serum phosphatidylcholine and triacylglycerol species were changed in patients with PM/DM compared to HI, suggesting disproportionate levels of saturated and polyunsaturated FAs that might have negative effects on muscle performance. After immunosuppressive treatment the total serum lipid levels of eicosadienoic (20:2, n-6) and eicosapentaenoic (20:5, n-3) acids were increased and serum phospholipid profiles were altered in patients with PM/DM. The correlation between FI or s-CK and levels of several lipid species indicate the important role of lipid changes in muscle performance and .Serum lipids profiles are significantly altered in patients with PM/DM compared to HI. Moreover, immunosuppressive treatment in patients newly diagnosed with PM/DM significantly affected serum lipid profiles. These findings provide new evidence of the dysregulated lipid metabolism in patients with PM/DM that could possibly contribute to low muscle performance.

Keyword: inflammation

and Oleic : The Yin and Yang of Fatty Acids in Type 2 Diabetes Mellitus.

Increased plasma non-esterified fatty acids (NEFAs) link obesity with insulin resistance and type 2 diabetes mellitus (T2DM). However, in contrast to the saturated FA (SFA) , the monounsaturated FA (MUFA) oleic elicits beneficial effects on insulin sensitivity, and the dietary :oleic ratio impacts diabetes risk in humans. Here we review recent mechanistic insights into the beneficial effects of oleic compared with on insulin resistance and T2DM, including its anti-inflammatory actions, and its capacity to inhibit endoplasmic reticulum (ER) stress, prevent attenuation of the insulin signaling pathway, and improve β cell survival. Understanding the molecular mechanisms of the antidiabetic effects of oleic may contribute to understanding the benefits of this FA in the prevention or delay of T2DM.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Gas chromatography coupled to mass spectrometry characterization, anti-inflammatory effect, wound-healing potential, and hair growth-promoting activity of Algerian L ().

The roots of have been used by the population of Northern Algeria to treat several pathological conditions, including wound healing and hair growth. The present study was conducted to evaluate the anti-inflammatory activity, wound-healing potential, and hair growth-promoting activity attributed to root.In this study, we have investigated the anti-inflammatory effect using carrageenan-induced paw edema test, evaluated the wound-healing potential by linear incision wound model, and evaluated hair growth activity using hair growth-promoting test attributed to root. Preliminary phytochemical screening and gas chromatography coupled to mass spectrometry (GC/MS) characterization were also performed.It was found that the methanolic extract of was characterized by the presence of tannins, flavonoids, anthocyanins, leucoanthocyanins, sennosides, free quinones, saponins, glycosides, mucilage, and coumarins. The GC/MS analysis could identify 22 compounds and showed that the major chemical constituents were (12.88%), mono(2-ethylhexyl) phthalate (12.75%), and 5-(hydroxymethyl)-2-furancarboxaldehyde, (9.19%). The phytoextract strongly inhibited ( < 0.001) paw edema formation in mice. The roots of also showed a significant ( < 0.05) wound-healing and hair growth-promoting effects.The results indicate the richness of the roots of the Algerian in biomolecules. These molecules exhibit an excellent reducing activity, a wound-healing property, and an interesting hair-promoting growth activity. All in all, the findings promote the usage of the Algerian as an effective and a safe potential skincare alternative remedy.

Keyword: inflammation

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated .

Keyword: inflammation

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: inflammation

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty metabolism was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. Lipid distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral lipid metabolic process" and "Regulation of lipid storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body energy metabolism.

Keyword: inflammation

Diabetes Mellitus-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and via Nuclear Mechanisms.

Objective- Macrophages play key roles in and diabetic vascular complications. Emerging evidence implicates long noncoding RNAs in , but their role in macrophage dysfunction associated with inflammatory diabetic complications is unclear and was therefore investigated in this study. Approach and Results- RNA-sequencing and real-time quantitative PCR demonstrated that a long noncoding RNA Dnm3os (dynamin 3 opposite strand) is upregulated in bone marrow-derived macrophages from type 2 diabetic db/db mice, diet-induced insulin-resistant mice, and diabetic ApoE mice, as well as in monocytes from type 2 diabetic patients relative to controls. Diabetic conditions (high glucose and ) induced Dnm3os in mouse and human macrophages. Promoter reporter analysis and chromatin immunoprecipitation assays demonstrated that diabetic conditions induce Dnm3os via NF-κB activation. RNA fluorescence in situ hybridization and real-time quantitative PCRs of subcellular fractions demonstrated nuclear localization and chromatin enrichment of Dnm3os in macrophages. Stable overexpression of Dnm3os in macrophages altered global histone modifications and upregulated and immune response genes and phagocytosis. Conversely, RNAi-mediated knockdown of Dnm3os attenuated these responses. RNA pull-down assays with macrophage nuclear lysates identified nucleolin and ILF-2 (interleukin enhancer-binding factor 2) as protein binding partners of Dnm3os, which was further confirmed by RNA fluorescence in situ hybridization immunofluorescence. Furthermore, nucleolin levels were decreased in diabetic conditions, and its knockdown enhanced Dnm3os-induced inflammatory gene expression and histone H3K9-acetylation at their promoters. Conclusions- These results demonstrate novel mechanisms involving upregulation of long noncoding RNA Dnm3os, disruption of its interaction with nucleolin, and epigenetic modifications at target genes that promote macrophage inflammatory phenotype in diabetes mellitus. The data could lead to long noncoding RNA-based therapies for inflammatory diabetes mellitus complications.

Keyword: inflammation

Human Milk and Donkey Milk, Compared to Cow Milk, Reduce Inflammatory Mediators and Modulate Glucose and Lipid Metabolism, Acting on Mitochondrial Function and Oleylethanolamide Levels in Rat Skeletal Muscle.

Milk from various species differs in nutrient composition. In particular, human milk (HM) and donkey milk (DM) are characterized by a relative high level of triacylglycerol enriched in in sn-2 position. These dietary fats seem to exert beneficial nutritional properties through N-acylethanolamine tissue modulation. The aim of this study is to compare the effects of cow milk (CM), DM, and HM on and glucose and lipid metabolism, focusing on mitochondrial function, efficiency, and dynamics in skeletal muscle, which is the major determinant of resting metabolic rate. Moreover, we also evaluated the levels of endocannabinoids and N-acylethanolamines in liver and skeletal muscle, since tissue fatty profiles can be modulated by nutrient intervention. To this aim, rats were fed with CM, DM, or HM for 4 weeks. Then, glucose tolerance and insulin resistance were analyzed. Pro-inflammatory and anti-inflammatory cytokines were evaluated in serum and skeletal muscle. Skeletal muscle was also processed to estimate mitochondrial function, efficiency, and dynamics, oxidative stress, and antioxidant/detoxifying enzyme activities. Fatty profiles, endocannabinoids, and N-acylethanolamine congeners were determined in liver and skeletal muscle tissue. We demonstrated that DM or HM administration reducing status, improves glucose disposal and insulin resistance and reduces lipid accumulation in skeletal muscle. Moreover, HM or DM administration increases redox status, and mitochondrial uncoupling, affecting mitochondrial dynamics in the skeletal muscle. Interestingly, HM and DM supplementation increase liver and muscle levels of the N-oleoylethanolamine (OEA), a key regulator of lipid metabolism and . HM and DM have a healthy nutritional effect, acting on inflammatory factors and glucose and lipid metabolism. This beneficial effect is associated to a modulation of mitochondrial function, efficiency, and dynamics and to an increase of OEA levels in skeletal muscle.

Keyword: inflammation

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

triggers inflammatory responses in N42 cultured hypothalamic cells partially via ceramide synthesis but not via TLR4.

A high-fat diet induces hypothalamic in rodents which, in turn, contributes to the development of obesity by eliciting both insulin and leptin resistance. However, the mechanism by which long-chain saturated fatty acids trigger is still contentious. To elucidate this mechanism, the effect of fatty acids on the expression of the pro-inflammatory cytokines IL-6 and TNFα was investigated in the mHypoE-N42 hypothalamic cell line (N42). N42 cells were treated with lauric (LA) and (PA). PA challenge was carried out in the presence of either a TLR4 inhibitor, a ceramide synthesis inhibitor (L-cycloserine), oleic (OA) or eicosapentaenoic (EPA). Intracellular ceramide accumulation was quantified using LC-ESI-MS/MS. PA but not LA upregulated IL-6 and TNFα. L-cycloserine, OA and EPA all counteracted PA-induced intracellular ceramide accumulation leading to a downregulation of IL-6 and TNFα. However, a TLR4 inhibitor failed to inhibit PA-induced upregulation of pro-inflammatory cytokines. In conclusion, PA induced the expression of IL-6 and TNFα in N42 neuronal cells independently of TLR4 but, partially, via ceramide synthesis with OA and EPA being anti-inflammatory by decreasing PA-induced intracellular ceramide build-up. Thus, ceramide accumulation represents one on the mechanisms by which PA induces in neurons.

Keyword: inflammation

Palmitate aggravates proteinuria-induced cell death and via CD36-inflammasome axis in the proximal tubular cells of obese mice.

High levels of serum free fatty acids (FFAs) and proteinuria have been implicated in the pathogenesis of obesity-related nephropathy. CD36, a class B scavenger receptor, is highly expressed in the renal proximal tubules and mediates FFA uptake. It is not clear whether FFA- and proteinuria-mediated CD36 activation coordinates NLRP3 inflammasomes to induce renal tubular injury and . In this study, we investigated the roles of CD36 and NLRP3 inflammasomes in FFA-induced renal injury in high-fat diet (HFD)-induced obesity. HFD-fed C57BL/6 mice and palmitate-treated HK2 renal tubular cells were used as in vivo and in vitro models. Immunohistochemical staining showed that CD36, IL-1β, and IL-18 levels increased progressively in the kidneys of HFD-fed mice. Sulfo- N-succinimidyl oleate (SSO), a CD36 inhibitor, attenuated the HFD-induced upregulation of NLRP3, IL-1β, and IL-18 and suppressed the colocalization of NLRP3 and ASC in renal tubular cells. In vitro, SSO abolished the palmitate-induced activation of IL-1β, IL-18, and caspase-1 in HK2 proximal tubular cells. Furthermore, treatment with SSO and the knockdown of caspase-1 expression by siRNA both inhibited palmitate-induced cell death and apoptosis in HK2 cells. Collectively, palmitate causes renal tubular , cell death, and apoptosis via the CD36/NLRP3/caspase-1 axis, which may explain, at least in part, the mechanism underlying FFA-related renal tubular injury. The blockade of CD36-induced cellular processes is therefore a promising strategy for treating obesity-related nephropathy.

Keyword: inflammation

Downregulation of miR-320 Alleviates Endoplasmic Reticulum Stress and Inflammatory Response in 3T3-L1 Adipocytes.

MicroRNAs serve important roles in the regulation of endoplasmic reticulum stress (ERs). This study aimed to investigate the role of microRNA-320 (miR-320) in the development of ERs and the inflammatory response in 3T3-L1 adipocytes.The adipose tissue expression levels of miR-320 and ERs markers (GRP78, GRP94, Derlin-1 and CHOP) and the serum concentration of inflammatory cytokines (TNF-α, NF-κB and IL-6) in obese patients were evaluated using quantitative real-time RT-PCR. The correlation of miR-320 with genes involved in ERs and inflammation was analyzed. The effects of miR-320 on ERs and inflammation were explored using mature 3T3-L1 adipocytes, which were pretreated with (PA).ERs markers and inflammatory cytokines were all upregulated in obese patients. Adipose tissue miR-320 expression was also increased in obese patients, and had positive correlations with the levels of ERs markers and inflammatory cytokines. After PA treatment, the levels of ERs markers and inflammatory cytokines were elevated significantly in 3T3-L1 adipocytes. Moreover, miR-320 expression was increased in the cells under ERs status. The upregulation of miR-320 could enhance the expression of ERs markers and inflammatory cytokines, but the downregulation of miR-320 resulted in the opposite results.The data of this study indicate that miR-320 expression is upregulated in ERs status, and the downregulation of miR-320 ameliorates ERs and the inflammatory response in 3T3-L1 adipocytes. We consider that the approaches to decrease miR-320 expression may be novel therapeutic strategies for the treatment of obesity and obesity-related diseases.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: inflammation

Sphingolipid changes do not underlie fatty -evoked GLUT4 insulin resistance nor signals in muscle cells.

Ceramides contribute to obesity-linked insulin resistance and in vivo, but whether this is a cell-autonomous phenomenon is debated, particularly in muscle, which dictates whole-body glucose uptake. We comprehensively analyzed lipid species produced in response to fatty acids and examined the consequence to insulin resistance and pro-inflammatory pathways. L6 myotubes were incubated with BSA-adsorbed palmitate or palmitoleate in the presence of myriocin, fenretinide, or fumonisin B1. Lipid species were determined by lipidomic analysis. Insulin sensitivity was scored by Akt phosphorylation and glucose transporter 4 (GLUT4) translocation, while pro-inflammatory indices were estimated by IκBα degradation and cytokine expression. Palmitate, but not palmitoleate, had mild effects on Akt phosphorylation but significantly inhibited insulin-stimulated GLUT4 translocation and increased expression of pro-inflammatory cytokines and Ceramides, hexosylceramides, and sphingosine-1-phosphate significantly heightened by palmitate correlated negatively with insulin sensitivity and positively with pro-inflammatory indices. Inhibition of sphingolipid pathways led to marked changes in cellular lipids, but did not prevent palmitate-induced impairment of insulin-stimulated GLUT4 translocation, suggesting that palmitate-induced accumulation of deleterious lipids and insulin resistance are correlated but independent events in myotubes. We propose that muscle cell-endogenous ceramide production does not evoke insulin resistance and that deleterious effects of ceramides in vivo may arise through ancillary cell communication.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: inflammation

The effect of enterolactone on liver lipid precursors of .

The aim of this study was to assess the effects of enterolactone (ENL) on lipid fractions fatty acids composition affecting hepatocyte development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free fatty acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their fatty acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of fatty acids were evaluated by Western Blot.Enterolactone modified fatty acids composition in FFA, DAG and TAG fractions. In conjunction with lipid overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated fatty acids in each of the investigated lipid fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: inflammation

Role of glutamine synthetase in angiogenesis beyond glutamine synthesis.

Glutamine synthetase, encoded by the gene GLUL, is an enzyme that converts glutamate and ammonia to glutamine. It is expressed by endothelial cells, but surprisingly shows negligible glutamine-synthesizing activity\xa0in these cells at physiological glutamine levels. Here we show in mice that genetic deletion of Glul in endothelial cells impairs vessel sprouting during vascular development, whereas pharmacological blockade of glutamine synthetase suppresses angiogenesis in ocular and inflammatory skin disease while only minimally affecting healthy adult quiescent endothelial cells. This relies on the inhibition of endothelial cell migration but not proliferation. Mechanistically we show that in human umbilical vein endothelial cells GLUL knockdown reduces membrane localization and activation of the GTPase RHOJ while activating other Rho GTPases and Rho kinase, thereby inducing actin stress fibres and impeding endothelial cell motility. Inhibition of Rho kinase rescues the defect in endothelial cell migration that is induced by GLUL knockdown. Notably, glutamine synthetase palmitoylates itself and interacts with RHOJ to sustain RHOJ palmitoylation, membrane localization and activation. These findings reveal that, in addition to the\xa0known formation of glutamine, the enzyme glutamine synthetase shows unknown\xa0activity in endothelial cell migration during pathological angiogenesis\xa0through RHOJ palmitoylation.

Keyword: inflammation

Molecular mechanism of activation of the immunoregulatory amidase NAAA.

Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and in animal models and in humans. Its signaling activity is terminated through degradation by -acylethanolamine amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand\'s acyl chain.

Keyword: inflammation

Downregulation of sirtuin 3 by increases the oxidative stress, impairment of mitochondrial function, and apoptosis in liver cells.

Elevated levels of saturated fatty acids show a strong cytotoxic effect in liver cells. Sirtuin 3 (SIRT3), a mitochondrially localized member of NAD -dependent deacetylase has been shown to protect hepatocytes against the oxidative stress. The role of SIRT3 on the cytotoxicity caused by fatty acids in liver cells is not fully understood. The aim of this study was to evaluate the expression level of SIRT3, oxidative stress, and mitochondrial impairments in human hepatoma HepG2 cells exposed to (PA). Our results showed that PA treatment caused the deposition of lipid droplets and resulted in an increased expression of tumor necrosis factor-α in a dose-dependent manner. Excessive accumulation of PA induces the reactive oxygen species formation and apoptosis while dissipating the mitochondrial transmembrane potential. The level of SIRT3 expression in both nuclear and mitochondrial fractions in HepG2 cells was decreased with the increase in PA concentrations. However, in the cytosolic fraction, the SIRT3 was undetectable. In conclusion, our results showed that PA caused an increase in and oxidative stress in HepG2 cells. The exposure of PA also resulted in the decline in transmembrane potential and an increase in apoptosis. The underexpression of nuclear and mitochondrial SIRT3 by PA suggests that the PA target the process that regulates the stress-related gene expression and mitochondrial functions.© 2019 Wiley Periodicals, Inc.

Keyword: inflammation

Synthesis of chemically edited derivatives of the endogenous regulator of 9-PAHSA.

Fatty esters of hydroxy fatty acids (FAHFAs) are a growing class of natural products found in organisms ranging from plants to humans. The roles these endogenous derivatives of fatty acids play in biology and their novel pathways for controlling have increased our understanding of basic human physiology. FAHFAs incorporate diverse fatty acids into their structures, however, given their recent discovery non-natural derivatives have not been a focus and as a result structure-activity relationships remain unknown. The importance of the long chain hydrocarbons extending from the ester linkage as they relate to anti-inflammatory activity is unknown. Herein the systematic removal of carbons from either the hydroxy fatty or fatty regions of the most studied FAHFA, ester of 9-hydroxystearic (9-PAHSA), was achieved and these synthetic, abridged analogs were tested for their ability to attenuate IL-6 production. Reduction of the carbon chain lengths of the 9-hydroxystearic portion or hydrocarbon chain resulted in lower molecular weight analogs that maintained anti-inflammatory activity or in one case enhanced activity.

Keyword: inflammation

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: inflammation

Chitosan Oligosaccharides Improve Glucolipid Metabolism Disorder in Liver by Suppression of Obesity-Related and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid metabolism disorder using (PA)-induced HepG2 cells and high-fat diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated lipid accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid metabolism-related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid metabolism disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid metabolism-related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid metabolism disorder by suppressing and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid metabolism-related diseases.

Keyword: inflammation

Elafibranor restricts lipogenic and inflammatory responses in a human skin stem cell-derived model of NASH.

Non-alcoholic steatohepatitis (NASH) is characterized by hepatocellular steatosis with concomitant hepatic . Despite its pandemic proportions, no anti-NASH drugs have been approved yet. This is partially because drug development is decelerated due to the lack of adequate tools to assess the efficacy of potential new drug candidates. The present study describes the development and application of a new preclinical model for NASH using hepatic cells generated from human skin-derived precursors. Exposure of these cells to lipogenic (insulin, glucose, fatty acids) and pro-inflammatory factors (IL-1β, TNF-α, TGF-β) resulted in a characteristic NASH response, as indicated by intracellular lipid accumulation, modulation of NASH-specific gene expression, increased caspase-3/7 activity and the expression and/or secretion of inflammatory markers, including CCL2, CCL5, CCL7, CCL8, CXCL5, CXCL8, IL1a, IL6 and IL11. The human relevance of the proposed NASH model was verified by transcriptomics analyses that revealed commonly modulated genes and the identification of the same gene classes between the in vitro system and patients suffering from NASH. The application potential of this in vitro model was demonstrated by testing elafibranor, a promising anti-NASH compound currently under clinical phase III trial evaluation. Elafibranor attenuated in vitro key features of NASH, and dramatically lowered lipid load as well as the expression and secretion of inflammatory chemokines, which in vivo are responsible for the recruitment of immune cells. This reduction in inflammatory response was NFκB-mediated. In summary, this human-relevant, in vitro system proved to be a sensitive testing tool for the investigation of novel anti-NASH compounds.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Hepatic fatty biosynthesis in KK-A mice is modulated by administration of persimmon peel extract: A DNA microarray study.

Previously, we showed that the intake of a persimmon peel (PP) extract altered hepatic gene expression associated with the insulin signaling pathway and enhanced tyrosine phosphorylation of insulin receptors in nonobese type 2 diabetic Goto-Kakizaki rats. Our objective was to evaluate the effect of fat-soluble PP extract on obese type 2 diabetic KK-A mice with insulin resistance.KK-A mice were fed a diet mixed with 0.1% of the extract for 8\xa0weeks. The total ketone body levels in the plasma of PP extract-fed mice were significantly lower than those in the normal diet-fed mice. Hepatic nonesterified content was higher in the PP extract-fed mice than in normal diet-fed mice. The hepatic gene expression profiles of the treated mice indicated upregulation of fatty synthesis and downregulation of -associated genes, predicting SREBP-1c and PPARγ activation.These results suggest that the PP extract enhances hepatic fatty synthesis via SREBP-1c and PPARγ, as well as anti-inflammatory activity in KK-A mice.

Keyword: inflammation

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of , fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver-body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, , enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: inflammation

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Metabolism Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and . At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic metabolism. DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: inflammation

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: inflammation

Ameliorative effect of panaxynol on the reduction in high-molecular-weight adiponectin secretion from 3T3-L1 adipocytes treated with acids.

Reduced plasma levels of the high-molecular weight (HMW) form of adiponectin, rather than total adiponectin levels, have been shown to be closely associated with various metabolic diseases including insulin resistance, type 2 diabetes, and cardiovascular disease. Therefore, we sought to explore active, naturally occurring compounds that promote the recovery of HMW adiponectin secretion suppressed by in our model. A total of 90 crude drug extracts were screened for the ability to augment HMW adiponectin secretion from 3T3-L1 adipocytes treated with . Panaxynol was isolated from Saposhnikovia divaricata as an active compound with HMW adiponectin promoting properties. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists are reported to increase the secretion of HMW adiponectin, although the effects of panaxynol were found to be independent of PPARγ activation. When the underlying mechanisms were further examined, panaxynol was found to inhibit the -induced downregulation of forkhead box O1 (FoxO1) protein, and the anti-lipotoxic effects were abolished by a FoxO1 inhibitor. Furthermore, CCAAT/enhancer-binding protein-α (C/EBPα) mRNA levels were also increased by panaxynol. Reactive oxygen species have critical roles in the reduction in HMW adiponection secretion by ; however, panaxynol reduced this increase in reactive oxygen species generation, followed by reductions in markers of endoplasmic reticulum stress and . Taken together, these findings suggest that panaxynol ameliorates the impaired HMW adiponection secretion in adipocytes treated with by restoring FoxO1 expression, owing to inhibition of reactive oxygen species generation, in a PPARγ-independent manner.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: inflammation

ShenFu Preparation Protects AML12 Cells Against -Induced Injury Through Inhibition of Both JNK/Nox4 and JNK/NFκB Pathways.

Nonalcoholic steatohepatitis includes steatosis along with liver , hepatocyte injury and fibrosis. In this study, we investigated the protective role and the potential mechanisms of a traditional Chinese medicine ShenFu (SF) preparation in an in vitro hepatic steatosis model.In (PA)-induced murine hepatic AML12 cell injury, effects of SF preparation on cellular apoptosis and intracellular triglyceride (iTG) level were assessed using TUNEL and TG Colorimetric Assay. Reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) levels were measured using DCF and JC-1 assay. Cytokine levels were evaluated using ELISA assay. Immunoblot was used to compare the activation level of c-Jun N terminal kinase (JNK), NADPH oxidase (Nox4), and NFκB pathways.Addition of SF preparation prevented PA-mediated increase of apoptosis and iTG as well as IL-8 and IL-6. In PA-treated cell, SF preparation reduced the level of Nox4 and ROS, while increasing the level of MMP and the expression of manganese superoxide dismutase (MnSOD) and catalase, indicating emendation of mitochondrial dysfunction. Nox4 inhibitor GKT137381 prevented PA-induced increase of ROS and apoptosis, while decreasing iTG slightly and not influencing the level of IL-8 and IL-6. SF preparation prevented PA-induced upregulation of phospho-JNK. JNK inhibitor SP600125 prevented PA-mediated increase of Nox4, IL-8, IL-6 and iTG. Nuclear translocation of NFκB/p65 was detected in PA-treated cells, which was prevented by SF preparation. An IκB degradation inhibitor, BAY11-7082, prevented PA-induced increase of IL-8 and IL-6 as well as iTG, whereas it only decreased ROS levels slightly and showed no influence on cellular apoptosis.SF preparation shows a beneficial role in prevention of hepatocyte injury by attenuating oxidative stress and cytokines production at least partially through inhibition of JNK/Nox4 and JNK/NFκB pathway, respectively.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: inflammation

Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether obesity impacts placental lipid transport and metabolism remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and insulin-sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal obesity. Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in lipid classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to insulin resistance and low-grade in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by obesity.-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

Keyword: inflammation

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic fatty liver disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, fat vacuoles and from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.Copyright © 2019. Published by Elsevier B.V.

Keyword: inflammation

Individual free fatty acids have unique associations with inflammatory biomarkers, insulin resistance and insulin secretion in healthy and gestational diabetic pregnant women.

We investigated the relationships of maternal circulating individual free fatty acids (FFA) with insulin resistance, insulin secretion and inflammatory biomarkers during mid-pregnancy.The data were drawn from a prospective cohort of generally healthy pregnant women (n=1368, African-American 36%, Hispanic 48%, Caucasian 16%) in Camden, NJ. We quantitatively determined 11 FFAs, seven cytokine/adipokine, homeostatic model assessment of insulin resistance (HOMA-IR) and C-peptide levels from the fasting blood samples that were collected at 16 weeks of gestation. Multivariate analyses were performed along with separate analyses for each individual FFA.High HOMA-IR (p<0.001) and C-peptide (p<0.0001) levels were positively associated with a twofold to fourfold increased risk for developing gestational diabetes mellitus (GDM). Negative relationships were found with specific FFAs (molecular percentage, palmitoleic, oleic, linolenic, myristic acids) and HOMA-IR and C-peptide levels (p<0.01\u2009to p<0.0001). In contrast, , stearic, arachidonic, dihomo-γ-linolenic (DGLA) and docosahexaenoic acids were positively associated with HOMA-IR and C-peptide (p<0.01\u2009to p<0.0001). The individual FFAs also predicted cytokine/adipokine levels. For example, women who had elevated DGLA (highest quartile) were twice as (adjusted OR 2.06, 95%\u2009CI 1.42 to 2.98) likely to have higher interleukin (IL)-8 (p<0.0001) levels. Conversely, women with high palmitoleic, oleic, and linolenic levels had reduced odds (≥2-fold, p<0.01\u2009to p<0.001) for having higher IL-8, IL-6 or tumor necrosis factor-alpha levels.Our results suggest that maternal individual FFAs uniquely affect insulin resistance and secretion. The effects are either direct or indirect via modulation of the inflammatory response. Modifying the composition of FFAs may help in reducing the risk of GDM.

Keyword: inflammation

[Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: inflammation

Thioacetamide potentiates high cholesterol and high fat diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH.

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to , fibrosis, collagen deposition were studied. The FTH mice featured hepatic , severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and liver anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: inflammation

A high-fat diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high-fat diet (HFD) commonly leads to obesity and insulin resistance. However, even 3\u2009days of HFD consumption has been linked to within the key homeostatic brain region, the hypothalamus.Mice were fed either a low-fat diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from fat for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose metabolism and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not . Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: inflammation

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: inflammation

Mechanisms of electrical remodeling in lipotoxic guinea pig heart.

To develop an adult guinea pig model of lipotoxicity and explore the underlying mechanisms associated with changes in the expression of the delayed rectifier potassium current (I).Lipotoxicity may represent a common link among metabolic disorders and a higher vulnerability to arrhythmias.Whole-cell patch clamp, and (PA, a potent inducer of lipotoxicity), were used to assess mechanisms of short-term (∼50 days) high-fat diet (HFD) feeding on atrial electrophysiology in guinea pig hearts and myocytes.HFD fed guinea pigs were significantly heavier, displayed hypertriglyceridemia and hypercholesterolemia; but no signs of hyperglycemia or compared to low-fat diet fed controls. Increasing cardiac PA levels, resulted in shortened atrial action potential duration, and increased I density. Inhibition of phosphoinositide 3-kinase (PI3K) prevented increases in I due to PA. Acute (≥1hr) exposure of atrial myocytes to exogenous PA (1\u202fmM) increased the density of the rapid delayed rectifier potassium current I, while it was decreased with the unsaturated oleic (OA, 1\u202fmM). Serine-threonine protein phosphatase-2 (PP2A) inhibition with cantharidin reversed the effect of OA on I.Our data provide evidence of a novel lipotoxic guinea pig model with signs of vulnerability to arrhythmias. Inhibition of PA/PI3K/I and/or activation of the OA/PP2A/I pathways may be therapeutically beneficial for lipotoxic arrhythmias.Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.

Keyword: inflammation

NDP-MSH reduces oxidative damage induced by in primary astrocytes.

Recent findings relate obesity to in key hypothalamic areas for body weight control. Hypothalamic has also been related to oxidative stress. (PA) is the most abundant free fatty found in food, and in vitro studies indicate that it triggers a pro-inflammatory response in the brain. Melanocortins are neuropeptides with proven anti-inflammatory and neuroprotective action mediated by melanocortin receptor 4 (MC4R), but little is known about the effect of melanocortins on oxidative stress. The aim of this study was to investigate whether melanocortins could alleviate oxidative stress induced by a high fat diet (HFD) model. We found that NDP-MSH treatment decreased PA-induced reactive oxygen species production in astrocytes, an effect blocked by the MC4R inhibitor JKC363. NDP-MSH abolished nuclear translocation of Nrf2 induced by PA and blocked the inhibitory effect of PA on superoxide dismutase (SOD) activity and glutathione levels while it also per se increased activity of SOD and γ-glutamate cysteine ligase (γ-GCL) antioxidant enzymes. However, HFD reduced hypothalamic MC4R and brain derived neurotrophic factor mRNA levels, thereby preventing the neuroprotective mechanism induced by melanocortins.© 2019 British Society for Neuroendocrinology.

Keyword: inflammation

Altered Lipidome Composition Is Related to Markers of Monocyte and Immune Activation in Antiretroviral Therapy Treated Human Immunodeficiency Virus (HIV) Infection and in Uninfected Persons.

HIV infection and antiretroviral therapy (ART) have both been linked to dyslipidemia and increased cardiovascular disease (CVD) risk. Alterations in the composition of saturated (SaFA), monounsaturated (MUFA), and polyunsaturated (PUFA) fatty acids are related to and CVD progression in HIV-uninfected (HIV-) populations. The relationships among the lipidome and markers of monocyte and immune activation in HIV-infected (HIV+) individuals are not well understood. Concentrations of serum lipids and their fatty composition were measured by direct infusion-tandem mass spectrometry in samples from 20 ART-treated HIV+ individuals and 20 HIV- individuals. HIV+ individuals had increased levels of free fatty acids (FFAs) with enrichment of SaFAs, including (16:0) and stearic (18:0), and these levels were directly associated with markers of monocyte (CD40, HLA-DR, TLR4, CD36) and serum (LBP, CRP). PUFA levels were reduced significantly in HIV+ individuals, and many individual PUFA species levels were inversely related to markers of monocyte activation, such as tissue factor, TLR4, CD69, and SR-A. Also in HIV+ individuals, the composition of lysophosphatidylcholine (LPC) was enriched for SaFAs; LPC species containing SaFAs were directly associated with IL-6 levels and monocyte activation. We similarly observed direct relationships between levels of SaFAs and in HIV uninfected individuals. Further, SaFA exposure altered monocyte subset phenotypes and inflammatory cytokine production . The lipidome is altered in ART-treated HIV infection, and may contribute to and CVD progression. Detailed lipidomic analyses may better assess CVD risk in both HIV+ and HIV- individuals than does traditional lipid profiling.

Keyword: inflammation

TERT inhibition leads to reduction of IL-6 expression induced by and interferes with the protective effects of tibolone in an astrocytic cell model.

Although it has been shown that telomerase has neuroprotective effects, mainly as a result of its non-canonical functions in neuronal cells, its role with respect to glial cells remains unknown. There is growing evidence indicating that telomerase plays an important role with respect to , especially in the regulation of pro-inflammatory cytokine gene expression. The present study aimed to evaluate the role of telomerase in an astrocyte cell model treated with (PA) and tibolone. Cell death, reactive oxygen species production and interleukin-6 expression were evaluated under telomerase inhibition with the BIBR1532 compound in T98G cells treated with tibolone and PA, using fluorometry, flow cytometry, enzyme-linked immunosorbent assays and the quantitative polymerase chain reaction. The results obtained showed that telomerase protein was increased by PA after 36\xa0hours, alone or in combination with tibolone, and that its activity was affected by PA. Telomerase inhibition reduced interleukin-6 expression and it interfered with the protective effects of tibolone on cell death. Moreover, tibolone increased Tyr707 phosphorylation in PA-treated cells. In the present study, we provide novel findings about the regulation of telomerase by PA and tibolone. Telomerase was involved in by PA and in protective effects of tibolone. Therefore, we conclude that telomerase could play a dual role in these cells.© 2019 British Society for Neuroendocrinology.

Keyword: inflammation

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: inflammation

METRNL attenuates lipid-induced and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, insulin resistance, and diabetes. Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: inflammation

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering but also produces uncanonical effects beyond such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the liver of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in liver cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the liver steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and liver lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: inflammation

Micronized palmitoylethanolamide reduces joint pain and glial cell activation.

Temporomandibular disorder (TMD) is a common painful condition in the temporomandibular joint (TMJ). Joint is believed to be a chief cause of pain in patients with TMD, through the release of pro-inflammatory cytokines that induce peripheral sensitization of nerve terminals followed by microglial stimulation.TMJ was induced in rats with the injection of complete Freund\'s adjuvant (CFA) emulsion into the left TMJ capsule.The present study would assess the effects of micronized palmitoylethanolamide (m-PEA) on glial activation and trigeminal hypersensitivity.Ten mg/kg m-PEA or corresponding vehicle was administered 1\xa0h after CFA and mechanical allodynia and edema were evaluated at 24 and 72\xa0h after CFA injection.CFA-injected animals showed TMJ edema and ipsilateral mechanical allodynia accompanied by a robust growth in GFAP protein-positive satellite glial cells and activation of resident macrophages in the TG. Moreover, m-PEA administration significantly reduced the degree of TMJ damage and pain, macrophage activation in TG and up-regulation of Iba1.The results confirm that m-PEA could represent a novel approach for monitoring pain during trigeminal nerve sensitization.

Keyword: inflammation

Anxa2 gene silencing attenuates obesity-induced insulin resistance by suppressing the NF-κB signaling pathway.

Insulin resistance (IR) continues to pose a major threat to public health due to its role in the pathogenesis of metabolic syndrome and its ever-increasing prevalence on a global scale. The aim of the current study was to investigate the efficacy of Anxa2 in obesity-induced IR through the mediation of the NF-κB signaling pathway. Microarray analysis was performed to screen differentially expressed genes associated with obesity. To verify whether Anxa2 was differentially expressed in IR triggered by obesity, IR mouse models were established in connection with a high-fat diet (HFD). In the mouse IR model, the role of differentially expressed Anxa2 in glycometabolism and IR was subsequently detected. To investigate the effect of Anxa2 on IR and its correlation with , a (PA)-induced IR cell model was established, with the relationship between Anxa2 and the NF-κB signaling pathway investigated accordingly. Anxa2 was determined to be highly expressed in IR. Silencing Anxa2 was shown to inhibit IR triggered by obesity. When Anxa2 was knocked down, elevated expression of phosphorylated insulin receptor substrate 1 (IRS1), IRS1 and peroxisome proliferator-activated receptor coactivator-1a, and glucose tolerance and insulin sensitivity along with 2-deoxy-d-glucose uptake was detected, whereas decreased expression of suppressor of cytokine signaling 3, IL-6, IL-1β, TNF-α, and p50 was observed. Taken together, the current study ultimately demonstrated that Anxa2 may be a novel drug strategy for IR disruption, indicating that Anxa2 gene silencing is capable of alleviating PA or HFD-induced IR and through its negative regulatory role in the process of p50 nuclear translocation of the NF-κB signaling pathway.

Keyword: inflammation

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free fatty acids (FFAs) are known to cause damage in various tissues by inducing . In this study, we investigated whether a FFA (palmitate) induces in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive oxygen species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish liver and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including diabetes.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Dietary modulation of energy homoeostasis and metabolic-.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy metabolism is a key determinant of . Whilst metabolic- is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: inflammation

Ferroptosis affects the progression of non-alcoholic steatohepatitis via the modulation of lipid peroxidation-mediated cell death in mice.

Oxidative stress and its-associated lipid peroxidation play a key role in non-alcoholic steatohepatitis (NASH). Ferroptosis is a recently recognized type of cell death characterized by an iron dependent and lipid peroxidation-mediated non-apoptotic cell death. We demonstrate the impact of ferroptosis on the progression of NASH induced by methionine/choline-deficient diet (MCD) feeding for 10 days. RSL-3 (a ferroptosis inducer) treatment showed decreased hepatic expression of GPX4, and conversely increased 12/15-lipoxygenase, and apoptosis inducing factor, indicating that ferroptosis plays a key role in NASH-related lipid peroxidation and its-associated cell death. Consistently, levels of serum biochemical, hepatic steatosis, , and apoptosis in MCD-fed mice were exacerbated with RSL-3 treatment. However, MCD-fed mice treated with sodium selenite (a GPX4 activator) showed increase of hepatic GPX4, accompanied by reduced NASH severity. To chelate iron, deferoxamine mesylate salt was used. Administration of deferoxamine mesylate salt significantly reduced NASH severity and abolished the harmful effects of RSL-3 in MCD-fed mice. Finally, treatment with Liproxstatin-1 (a ferroptosis inhibitor) repressed hepatic lipid peroxidation, and its associated cell death, resulting in decreased NASH severity. Consistent with the in vivo findings, modulation of ferroptosis /GPX4 affected hepatocellular death in -induced in vitro NASH milieu. We conclude that GPX4 and its-related ferroptosis might play a major role in the development of NASH.Copyright © 2019. Published by Elsevier Inc.

Keyword: inflammation

TAp63 is correlated with chronic in patients with newly diagnosed type 2 diabetes mellitus.

To investigate TAp63 expression in patients with type 2 diabetes mellitus (T2DM) and the potential correlations between TAp63 and proinflammatory cytokines production and other clinical parameters.Peripheral blood mononuclear cells (PBMCs) and plasma were collected from 72 T2DM (cases) and 72 healthy subjects (controls). Fasting blood glucose (FBG), fasting insulin (FIN) and a blood lipid profile were measured. The homeostasis model assessment (HOMA) was used to estimate insulin resistance (IR). Plasma tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were determined. PBMCs isolated from healthy subjects were cultured with or without 33.3 mmol/l glucose or 0.5 mmol/l (PA) for 6 h, 24 h, 48 h, and 72 h. The expression of TAp63 at mRNA and protein levels in PBMCs was analyzed using real-time qRT-PCR and western blots, respectively.TAp63 expression was significantly lower in T2DM patients compared with that of the controls. In addition, TAp63 expression showed a negative correlation with FBG, FIN, HbA1c, HOMA-IR, FFAs, TNF-α, and IL-6 levels. Treatment with 33.3 mmol/l glucose or 0.5 mmol/l PA increased TAp63 expression in the cultured PBMCs.TAp63 level may be correlated with chronic inflammatory state and perturbed glucose and lipid metabolism in T2DM.Copyright © 2018. Published by Elsevier Inc.

Keyword: inflammation

Hydroxystearic Acids Activate GPR40, Which Is Involved in Their Beneficial Effects on Glucose Homeostasis.

hydroxystearic acids (PAHSAs) are endogenous lipids with anti-diabetic and anti-inflammatory effects. PAHSA levels are reduced in serum and adipose tissue of insulin-resistant people and high-fat diet (HFD)-fed mice. Here, we investigated whether chronic PAHSA treatment enhances insulin sensitivity and which receptors mediate PAHSA effects. Chronic PAHSA administration in chow- and HFD-fed mice raises serum and tissue PAHSA levels ∼1.4- to 3-fold. This improves insulin sensitivity and glucose tolerance without altering body weight. PAHSA administration in chow-fed, but not HFD-fed, mice augments insulin and glucagon-like peptide (GLP-1) secretion. PAHSAs are selective agonists for GPR40, increasing Ca flux, but not intracellular cyclic AMP. Blocking GPR40 reverses improvements in glucose tolerance and insulin sensitivity in PAHSA-treated chow- and HFD-fed mice and directly inhibits PAHSA augmentation of glucose-stimulated insulin secretion in human islets. In contrast, GLP-1 receptor blockade in PAHSA-treated chow-fed mice reduces PAHSA effects on glucose tolerance, but not on insulin sensitivity. Thus, PAHSAs activate GPR40, which is involved in their beneficial metabolic effects.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Amelioration of Endoplasmic Reticulum Stress by Mesenchymal Stem Cells via Hepatocyte Growth Factor/c-Met Signaling in Obesity-Associated Kidney Injury.

Recent advances in the understanding of lipid metabolism suggest a critical role of endoplasmic reticulum (ER) stress in obesity-induced kidney injury. Hepatocyte growth factor (HGF) is a pleiotropic cytokine frequently featured in stem cell therapy with distinct renotropic benefits. This study aims to define the potential link between human induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs)/bone marrow-derived MSCs (BM-MSCs) and ER stress in lipotoxic kidney injury induced by (PA) in renal tubular cells and by high-fat diet (HFD) in mice. iPS-MSCs or BM-MSCs alleviated ER stress (by preventing induction of Bip, chop, and unfolded protein response), (Il6, Cxcl1, and Cxcl2), and apoptosis (Bax/Bcl2 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells) in renal cortex of animals exposed to HFD thus mitigating histologic damage and albuminuria, via activating HGF/c-Met paracrine signaling that resulted in enhanced HGF secretion in the glomerular compartment and c-Met expression in the tubules. Coculture experiments identified glomerular endothelial cells (GECs) to be the exclusive source of glomerular HGF when incubated with either iPS-MSCs or BM-MSCs in the presence of PA. Furthermore, both GEC-derived HGF and exogenous recombinant HGF attenuated PA-induced ER stress in cultured tubular cells, and this effect was abrogated by a neutralizing anti-HGF antibody. Taken together, this study is the first to demonstrate that MSCs ameliorate lipotoxic kidney injury via a novel microenvironment-dependent paracrine HGF/c-Met signaling mechanism to suppress ER stress and its downstream pro-inflammatory and pro-apoptotic consequences. Stem Cells Translational Medicine 2019;8:898&910.© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

Keyword: inflammation

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic , lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: inflammation

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis (NASH) is characterized by lipid accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on NASH and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce NASH and liver fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated lipid accumulation, and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents NASH and fibrosis via inhibition of oxidative stress and , highlighting polydatin as a potential therapeutic agent for prevention and treatment of NASH.

Keyword: inflammation

Nicotinic Receptor GPR109A Exerts Anti-Inflammatory Effects Through Inhibiting the Akt/mTOR Signaling Pathway in MIN6 Pancreatic β cells.

We found that activation of the nicotinic receptor GPR109A, expressed by the MIN6 murine pancreatic β cell line, inhibits nitric oxide accumulation induced by IFN-γ and TNF-α, implicating an anti-inflammatory effect of GPR109A in MIN6 cells. Nevertheless, the mechanism of its anti-inflammatory effect is still unknown. In this study, we used to stimulate MIN6 cells to induce inflammatory cytokine production and explored the mechanism by which GPR109A exerts anti-inflammatory effects.RT-PCR and immunocytochemical staining were used to detect the expression of GPR109A in MIN6 cells. Western blotting was used to detect the activation of the Akt/mTOR signaling pathway and expression of the inflammatory cytokine INF-γ, in MIN6 cells, following treatments with and +nicotinic , or with different concentrations of nicotinic and 3-hydroxybutyrate.In MIN6 cells, GPR109A transcripts and protein are expressed and GPR109A protein is mainly located in the cell membrane and cytoplasm. enhanced the phosphorylation of Akt and p70S6K and elevated the expression of IFN-γ. Co-treatment with nicotinic , which is an agonist of GPR109A, inhibited the -induced phosphorylation of Akt, mTOR, and p70S6K, as well as the expression of IFN-γ.GPR109A may inhibit inflammatory cytokine production, induced by , by MIN6 cells possibly via inhibiting the Akt/mTOR signaling pathway.© 2017 by the Association of Clinical Scientists, Inc.

Keyword: inflammation

Clinopodium chinense Attenuates -Induced Vascular Endothelial and Insulin Resistance through TLR4-Mediated NF- B and MAPK Pathways.

Elevated (PA) levels are associated with the development of , insulin resistance (IR) and endothelial dysfunction. Clinopodium chinense (Benth.) O. Kuntze has been shown to lower blood glucose and attenuate high glucose-induced vascular endothelial cells injury. In the present study we investigated the effects of ethyl acetate extract of C. chinense (CCE) on PA-induced and IR in the vascular endothelium and its molecular mechanism. We found that CCE significantly inhibited PA-induced toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs). Consequently, this led to the inhibition of the following downstream adapted proteins myeloid differentiation primary response gene 88, Toll/interleukin-1 receptor domain-containing adaptor-inducing interferon- and TNF receptor-associated factor 6. Moreover, CCE inhibited the phosphorylation of Ikappa B kinase , nuclear factor kappa-B (NF- B), c-Jun N-terminal kinase, extracellular regulated protein kinases, p38-mitogen-activated protein kinase (MAPK) and subsequently suppressed the release of tumor necrosis factor- , interleukin-1 (IL-1 ) and IL-6. CCE also inhibited IRS-1 serine phosphorylation and ameliorated insulin-mediated tyrosine phosphorylation of IRS-1. Moreover, CCE restored serine/threonine kinase and endothelial nitric oxide synthase (eNOS) activation and thus increased insulin-mediated nitric oxide (NO) production in PA-treated HUVECs. This led to reverse insulin mediated endothelium-dependent relaxation, eNOS phosphorylation and NO production in PA-treated rat thoracic aortas. These results suggest that CCE can significantly inhibit the inflammatory response and alleviate impaired insulin signaling in the vascular endothelium by suppressing TLR4-mediated NF- B and MAPK pathways. Therefore, CCE can be considered as a potential therapeutic candidate for endothelial dysfunction associated with IR and diabetes.

Keyword: inflammation

Tetrahydroxy stilbene glucoside alleviates -induced and apoptosis in cardiomyocytes by regulating miR-129-3p/Smad3 signaling.

Tetrahydroxy stilbene glucoside (TSG) has been reported to exert a cytoprotective effect against various toxicants. However, the function and mechanism of TSG in (PA)-induced and apoptosis in cardiomyocytes are still unknown. The present study was designed to investigate the post-transcriptional mechanism in TSG-treated cardiomyocytes\' and apoptosis induced by PA.The mRNA and protein levels were assayed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining was used to evaluate apoptosis using flow cytometry.TSG restricted the detrimental effects, including the activated inflammatory response and apoptosis, of PA in cardiomyocytes, as well as the up-regulation of miR-129-3p and down-regulation of p-Smad3 expression. In addition, bioinformatics and experimental analysis suggested that Smad3 was a direct target of miR-129-3p, which could inhibit or enhance the expression of p-Smad by transfection with miR-129-3p mimics or inhibitors, respectively. Furthermore, our results demonstrated that overexpression of Smad3 reversed the inhibition of and apoptosis by overexpression of miR-129-3p in PA-stimulated cardiomyocytes.TSG targeted to miR-129-3p/Smad3 signaling inhibited PA-induced and apoptosis in cardiomyocytes.

Keyword: inflammation

Untargeted metabolomic analysis of coronary artery disease patients with diastolic dysfunction show disturbed oxidative pathway.

Left ventricular diastolic dysfunction (LVDD) is common in patients with coronary artery disease (CAD) with prevalence estimates of 34% and constitutes a predictor of all-cause mortality. Although diastolic dysfunction is induced by myocardial ischemia and has been shown to alter the clinical course, the role of coronary artery disease in the diastolic dysfunction and its progression into heart failure has not been completely elucidated.The present study was conducted to identify possible metabolites in coronary artery disease patients that are differentially regulated in patients with diastolic dysfunction.The serum of CAD (n\u2009=\u200975) patients and young healthy volunteers (n\u2009=\u200943) were analysed by using gas chromatography mass spectrometry (GC-MS) technique. Pre-processing of data results in 1547 features; among them 1064 features were annotated using NIST library.Fifteen metabolites were found to be statistically different between cases and control. Variation in metabolites were identified and correlated with several clinically important echocardiography parameters i.e. LVDD grades, ejection fraction (EF) and E/e\' values. The results suggested that metabolic products of fatty oxidation and glucose oxidation pathways such as oleic , stearic , , linoleic , galactose, pyruvic and lactic acids are predominantly up regulated in patients with coronary artery disease and severity of diastolic dysfunction appears to be linked to increase in fatty oxidation and . The metabolic fingerprints of these patients give us an insight into the pathophysiological mechanism of diastolic dysfunction in coronary artery disease patients although it did not identify validated novel markers.

Keyword: inflammation

Low molecular weight fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic fatty-liver disease (NAFLD), caused by elevated hepatic lipids, and oxidative stress, is the most common liver disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in diabetes-induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the liver of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and . Therefore, LMWF provides a potential supplementary treatment for obesity/diabetes-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

Oleic protects saturated fatty mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis.

Aim This study aims to demonstrate the protective effects of monounsaturated oleic (OA) against saturated (PA) induced cellular lipotoxicity in hepatocytes and rats with non-alcoholic steatohepatitis (NASH).Human hepatoma cell line HepG2 cells and neonatal rat primary hepatocytes were treated with PA or/and OA for 24\u202fh. SD rats were fed with high fat diet (HFD) to induce NASH. From the 16th w, the HFD was full or half replaced by olive oil to observe the protective effects.In vitro, OA substantially alleviated PA induced cellular apoptosis, oxidative stress, ER stress, mitochondrial dysfunction, as well as in hepatocytes. In vivo, only olive oil supplementation had no detrimental effects, while HFD developed NASH in normal rats. Full replacement of HFD with olive oil had profoundly reversed NASH. Noteworthily, half replacement of HFD with olive oil (a mixed diet) has ameliorated NASH injury as well. It strikingly changed the hepatic histology from macrovesicular-steatosis into entire microvesicular-steatosis, and significantly reduced , ballooning and fibrosis.Our study has demonstrated in both hepatocytes and NASH rats that oleic acids had great potential to combat the saturated fatty acids induced hepatic lipotoxicity. Only half replacement of HFD by monounsaturated fatty acids rich diet still had significant therapeutic outcome in NASH rats. Redirecting the toxic saturated fatty acids into triglyceride storage and reduction of cholesterol accumulation might be the possible explanation of OA driven protection in this scenario.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Investigation of the phytochemical composition and antioxidant properties of chinar (Platanus orientalis L.) leaf infusion against ethanol-induced oxidative stress in rats.

Chinar (Platanus orientalis L.) is used in folk medicine against tooth and knee pain, wounds, , and stomach discomfort; however, the effects of P. orientalis leaf (PO-leaf) infusion on the liver and kidney are unknown. The aim of this study was to investigate the phytochemical composition and antioxidant properties of an infusion obtained from dried P. orientalis leaves against ethanol-induced oxidative stress (OS) in rats. After a toxicity test, thirty male Wistar rats were divided into five groups: Control, Ethanol 20%, Ethanol 20%\u2009+\u2009Silymarin (10\xa0mg/kg), Ethanol 20%\u2009+\u2009PO-20\xa0mg/mL infusion, and Ethanol 20%\u2009+\u2009PO-60\xa0mg/mL infusion. The PO-leaf infusion doses were given ad libitum during 28\xa0days to test the biochemical and antioxidant enzyme levels. According to the results, the PO-leaf contained rich compounds such as benzaldehyde, , 2,4-ditert-butylphenol, stearic , octadecanoic , linoleic , linolenic , kaempferol, and kaempferol derivatives. In the Ethanol group, AST, ALT, LDH, GGT, UA, and urea in the serum and GST and malondialdehyde (MDA) in the liver and erythrocyte tissues showed a significant increase compared to the Control group. AST, LDH, GGT, UA, and LDL-C levels in the serum and MDA (all tissues) significantly decreased in the Ethanol\u2009+\u2009PO-60\xa0mg/mL group compared to the Ethanol group. SOD, GPx, and CAT activities in the kidney tissue of the Ethanol group showed a significant decrease compared to the Control group, whereas the GPx activity in kidney tissue in all of the treatment groups increased significantly compared to the Ethanol group. These findings suggest that the administration of the determined PO-leaf infusion doses might have a protective role against ethanol-induced liver and kidney damage in rats.

Keyword: inflammation

Benefits of sea buckthorn (Hippophae rhamnoides) pulp oil-based mouthwash on oral health.

The purpose of this study was to conduct phytochemical analysis of sea buckthorn pulp oil and to evaluate the antimicrobial, anti-biofilm and antioxidant activities of its mouthwash form.Fatty composition of the sea buckthorn pulp oil was determined by GC-MS analysis, which revealed that, mono-unsaturated fatty , palmitoleic and saturated fatty , , were the major constituents. The antimicrobial and the anti-biofilm capacities of sea buckthorn pulp oil mouthwash form were evaluated against Streptococcus gordonii, Porphyromonas gingivalis, Actinomyces viscosus and Candida albicans, according to the European Norms, and the Biofilm Ring Test , respectively. These activities were then compared with those of chlorhexidine and herbal mouthwashes. The sea buckthorn-based mouthwash was bactericidal against S. gordonii and P. gingivalis, bacteriostatic against A. viscosus and showed no antifungal effect. Regardless of the strains used, complete inhibition of biofilm formation was achieved. The antioxidant activity of this experimental mouthwash was also assessed by DPPH and NBT assays.Sea buckthorn mouthwash showed anti-biofilm activities against select single and multiple oral bacterial species.In this study, a mouthwash derived from sea buckthorn (Hippophae rhamnoides) pulp oil has been experimented, for the first time, in order to overcome the problem of a large number of available synthetic mouthwashes which have side effects on teeth, gums and mucous membranes. This mouthwash seemed to be a suitable alternative for a preventive agent for periodontal .© 2019 The Society for Applied Microbiology.

Keyword: inflammation

Saturated fatty combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic , a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: inflammation

Lipotoxic Effects of on Astrocytes Are Associated with Autophagy Impairment.

Obesity is associated with an increase in the brain levels of saturated free fatty acids, such as (PA). Previous studies have shown that PA exerts proinflammatory actions and reduces cell viability in astrocyte cultures. In this study, we have assessed whether an alteration in autophagy is involved in the effects of PA on astrocytes. Primary astrocytes were obtained from the cerebral cortex of male and female CD1 mouse pups and were incubated for 4.5 or 24\xa0h with 250-500\xa0μM PA. PA increased the levels of LC3-II, an autophagosome marker, and reduced LC3-II flux in astrocytes, suggesting a blockade of autophagy. This effect was observed both after 4.5 and 24\xa0h of treatment with PA. PA had additional effects after treatment for 24\xa0h, increasing the expression of proinflammatory cytokines, decreasing cell viability, and increasing the levels of an endoplasmic reticulum stress marker. In addition, PA decreased the expression of estrogen receptors, but only in female astrocytes. However, the treatment with estradiol, estrogen receptor agonists, or inhibitor of estradiol synthesis did not counteract the action of PA on cell viability. Rapamycin, an autophagy inducer, was unable to prevent the effect of PA on cell viability. In addition, hydroxychloroquine, an autophagy blocker, did not cause per se astrocyte death. These findings suggest that the effect of PA on autophagy is not sufficient to induce astrocyte loss, which is only observed when prolonged PA treatment causes other alterations in astrocytes, such as increased and endoplasmic reticulum stress.

Keyword: inflammation

Overexpression of heart-type fatty binding protein enhances fatty -induced podocyte injury.

Deregulated lipid metabolism is a characteristic of metabolic diseases including type 2 diabetes and obesity, and likely contributes to podocyte injury and end-stage kidney disease. Heart-type fatty binding protein (H-FABP) was reported to be associated with lipid metabolism. The present study investigated whether H-FABP contributes to podocyte homeostasis. Podocytes were transfected by lentiviral vector to construct a cell line which stably overexpressed H-FABP. Small interfering RNA capable of effectively silencing H-FABP was introduced into podocytes to construct a cell line with H-FABP knockdown. Certain groups were treated with (PA) and the fat metabolism, as well as inflammatory and oxidative stress markers were measured. PA accelerated lipid metabolism derangement, inflammatory reaction and oxidative stress in podocytes. Overexpression of H-FABP enhanced the PA-induced disequilibrium in podocytes. The mRNA and protein expression levels of acyl-coenzyme A oxidase 3 and monocyte chemotactic protein 1, and the protein expression levels of 8-hydroxy-2\'-deoxyguanosine and 4-hydroxynonenal were upregulated in the H-FABP overexpression group, while the mRNA and protein expression of peroxisome proliferator activated receptor α was downregulated. Knockdown of H-FABP inhibited the PA-induced injury and lipid metabolism derangement, as well as the inflammatory reaction and oxidative stress in podocytes. These results indicated that overexpression of H-FABP enhances fatty -induced podocyte injury, while H-FABP inhibition may represent a potential therapeutic strategy for the prevention of lipid metabolism-associated podocyte injury.

Keyword: inflammation

Wnt7a promotes wound healing by regulation of angiogenesis and : Issues on diabetes and obesity.

Diabetic skin heals wounds poorly. Though obesity is the common risk factor of diabetes mellitus, few studies have investigated its effects on wound healing.This study aimed to evaluate the morphology and possible mechanism of human umbilical vein endothelial cells (HUVEC-C) in response to different levels of glucose and , and explore the role of Wnt7a in wound healing.The functional changes of HUVEC-C and mRNA expression of Wnt signaling were determined by analyzing cell viability, migration, tube formation and rt-PCR in gradients of glucose and . Recombinant Wnt7a protein was injected around wounds made on streptozotocin (STZ) -induced diabetic rats with (HF) or without (DM) high-fat diet. Angiogenesis and inflammatory statement were mainly analyzed by immunohistochemistry, ELISA, cytometry and Western blotting.The expression of Wnt7a significantly decreased in high Glc/PA cultured cells or DM and HF wounded rats. Impaired wound healing was also observed in DM and HF groups. The healing rate significantly accelerated after localized injection with Wnt7a at d10. Moreover, the expression of CD31, eNOS phosphorylation and NO were increased; the reduction of local neutrophils influx, ICAM-1 and IL-6/8 expression levels were obvious especially in diabetic with obesity rats at d10 after Wnt7a treatment.This study indicates the potential role of Wnt7a, which is beneficial for regeneration of damaged vessels, moderation of inflammatory statement in diabetic wound healing with or without obesity, thus demonstrating its possible utility as a topical administration to promote healing rate.Copyright © 2018. Published by Elsevier B.V.

Keyword: inflammation

Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic fatty liver to nonalcoholic steatohepatitis.

Activation of is an important mechanism in the development of nonalcoholic steatohepatitis (NASH). This study aims to delineate how mitophagy affects NLRP3 inflammasome activation in hepatic lipotoxicity. Mice were fed a high fat/calorie diet (HFCD) for 24 weeks. Primary rat hepatocytes were treated with (PA) for various periods of time. Mitophagy was measured by protein levels of LC3II and P62. NLRP3, caspase-1, interleukin (IL)-18, and IL-1β at mRNA and protein levels were used as indicators of inflammasome activation. Along with steatotic progression in HFCD-fed mice, ratio of LC3II/β-actin was decreased concurrently with increased levels of liver P62, NLRP3, caspase-1, IL-1β, IL-18, and serum IL-1β levels in late-stage NASH. PA treatment resulted in mitochondrial oxidative stress and initiated mitophagy in primary hepatocytes. The addition of cyclosporine A did not change LC3II/Τοmm20 ratios; but P62 levels were increased after an extended duration of PA exposure, indicating a defect in autophagic activity. Along with impaired mitophagy, mRNA and protein levels of NLRP3, caspase-1, IL-18 and IL-1β were upregulated by PA treatment. Pretreatment with MCC950, N-acetyl cysteine or acetyl-L-carnitine reversed inflammasome activation and a pyroptotic cascade. Additionally, mitophagic flux was partially recovered as indicated by increases in LC3II/Tomm20 ratio, parkin, and PINK1 expression, and decreased P62 expression. The findings suggest that impaired mitophagy triggers hepatic NLRP3 inflammasome activation in a murine NASH model and primary hepatocytes. The new insights into inflammasome activation through mitophagy advance our understanding of how fatty acids elicit lipotoxicity through oxidant stress and autophagy in mitochondria.

Keyword: inflammation

Caveolin-1 prevents palmitate-induced NF-κB signaling by inhibiting GPRC5B-phosphorylation.

Tyrosine phosphorylation of GPRC5B and phosphorylation-dependent recruitment of Fyn through the SH2 domain have been implicated in NF-κB activation and obesity-linked adipose . GPRC5B tightly associates with caveolin-1 (Cav1); however, the role of this interaction remains elusive. Here, we report that Cav1 reduces GPRC5B-mediated NF-κB signaling by blocking GPRC5B-phosphorylation. We demonstrate highly abundant tyrosine phosphorylation of GPRC5B is observed in Neuro2a cells lacking endogenous Cav1 expression. Reversely, exogenous expression of Cav1 in these cells inhibits GPRC5B-phosphorylation. Although GPRC5B lacks conventional caveolin-binding motif, cytoplasmic tail of GPRC5B directly interacts with the C-terminal domain of Cav1. The vacant scaffolding domain of Cav1 in the protein complex suggests a potential mechanism for blocking GPRC5B-phosphorylation by Cav1, because Fyn loses the activity by binding with Cav1-scaffolding domain. Enhanced GPRC5B-mediated NF-κB signaling in Cav1-deficient cells were observed under palmitate-induced metabolic stress. These results support Cav1 functions as a negative modulator for GPRC5B action.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Observational clinical and nerve conduction study on effects of a nutraceutical combination on painful diabetic distal symmetric sensory-motor neuropathy in patients with diabetes type 1 and type 2.

Painful distal symmetric polyneuropathy (pDSPN) is one of the most common and invalidating complications of diabetes mellitus, both of type 1 and type 2. Mechanisms responsible for the occurrence of the pDSPN are multifactorial and involve metabolic pathways regulating , microvessel circulation, axonal degeneration and so on. Several therapeutic approaches have been proposed to treat pain and each of them showed positive effects associated to drug-related side effects.Twenty-five consecutive patients with diagnosis of diabetes mellitus and pDSPN and tried to manage pain with a dietary supplement composed of a mixture of natural extracts (β-caryophyllene, myrrh, carnosic ) and PEA. This is a nutraceutical with potential multiple effects on metabolic, pain and vascular compartments, a profile considered useful in pDSPN. Patients were enrolled and polyneuropathy evaluated by means of nerve conduction study. Pain was assessed using VAS score scale and MNSI. Each patient was evaluated at T0 (time of enrollment) and at T1 (after 16 weeks of treatment).Supplement administration was well tolerated and induced unexpectedly significant amelioration of polyneuropathy with increase amplitude and reduction of pain. No side effects were reported.This fixed combination could well be considered as a potential nutraceutical option to manage pDSPN in diabetic patients.

Keyword: inflammation

Pharmacological activities of the organic extracts and fatty composition of the petroleum ether extract from Haplophyllum tuberculatum leaves.

Haplophyllum tuberculatum is used in traditional medicine to treat many disorders including and pain. The aim of this study is to investigate the organic extracts from H. tuberculatum leaves against , gastric ulcer and pain.Acute toxicity was studied in vivo to determine the toxic doses of the organic extracts. Anti-inflammatory activity was also evaluated in vivo using carrageenan-induced paw edema in Wistar rats. Gastroprotective activity was tested using the HCl/ethanol-induced gastric ulcer test in rats. Peripheral and central analgesic activities were assessed using the acetic -induced writhing test and the hot-plate method, respectively. The chemical composition of the fatty acids in the petroleum ether (PE) extract was determined with GC-MS.At 25, 50 and 100mg/kg PE extract was the most active against . Percentages inhibition 5h after carrageenan-injection were 51.12; 86.71% and 96.92%, respectively. The same extract at 100mg/kg showed good analgesic activities using the acetic -induced writhing test and the hot-plate method. The chloroform, ethyl acetate (EtOAc) and butanolic (n-BuOH) extracts exhibited strong anti-inflammatory, gastroprotective and analgesic activities at 100mg/kg. The GC-FID analysis revealed that the PE extract was rich in γ-linolenic (45.50%) followed by (18.48%), linoleic (10.73%), erucic (4.72), stearic (3.96%) and oleic (2.57%).The results of the present study support the traditional use of the leaves of H. tuberculatum and may possibly serve as prospective material for further development of safe new phytochemical anti-inflammatory, gastroprotective and/or analgesic agents.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

N-3 Polyunsaturated Fatty Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Diabetes Rats by Postponing Shortening of Hepatic Telomeres and Modulating Liver Metabolism.

The long-term influence of gestational diabetes mellitus (GDM) on offspring and the effect of omega-3 polyunsaturated fatty acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and in the liver of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the liver, in which numerous metabolites and metabolic pathways were altered when GDM offspring grew to old age. Many metabolites were related to diabetes risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious diabetes risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: inflammation

Hybrid lipids, peptides, and lymphocytes: new era in type 1 diabetes research.

Type 1 diabetes (T1D) results from autoimmune destruction of insulin-producing β cells in islets of Langerhans. Many genetic and immunological insights into autoimmune disease pathogenesis were initially uncovered in the context of T1D and facilitated by preclinical studies using the nonobese diabetic (NOD) mouse model. Recently, the study of T1D has led to the discovery of fatty esters of hydroxyl fatty acids (FAHFAs), which are naturally occurring hybrid peptides that modulate and diabetes pathogenesis, and a hybrid lymphocyte that expresses both B and T cell receptors. esters of hydroxy stearic acids (PAHSAs) are the most extensively studied FAHFA. In this issue of the JCI, Syed et al. have shown that PAHSAs both attenuate autoimmune responses and promote β cell survival in NOD mice. Given the lack of effective T1D therapies and the paucity of known side effects of PAHSAs, this lipid may have therapeutic potential for individuals at risk for or newly diagnosed with T1D.

Keyword: inflammation

Fatty Metabolism is Associated With Disease Severity After H7N9 Infection.

Human infections with the H7N9 virus could lead to lung damage and even multiple organ failure, which is closely associated with a high mortality rate. However, the metabolic basis of such systemic alterations remains unknown.This study included hospitalized patients (n\u202f=\u202f4) with laboratory-confirmed H7N9 infection, healthy controls (n\u202f=\u202f9), and two disease control groups comprising patients with pneumonia (n\u202f=\u202f9) and patients with pneumonia who received steroid treatment (n\u202f=\u202f10). One H7N9-infected patient underwent lung biopsy for histopathological analysis and expression analysis of genes associated with lung homeostasis. H7N9-induced systemic alterations were investigated using metabolomic analysis of sera collected from the four patients by using ultra-performance liquid chromatography-mass spectrometry. Chest digital radiography and laboratory tests were also conducted.Two of the four patients did not survive the clinical treatments with antiviral medication, steroids, and oxygen therapy. Biopsy revealed disrupted expression of genes associated with lung epithelial integrity. Histopathological analysis demonstrated severe lung after H7N9 infection. Metabolomic analysis indicated that fatty metabolism may be inhibited during H7N9 infection. Serum levels of , erucic , and phytal may negatively correlate with the extent of lung after H7N9 infection. The changes in fatty levels may not be due to steroid treatment or pneumonia.Altered structural and secretory properties of the lung epithelium may be associated with the severity of H7N9-infection-induced lung disease. Moreover, fatty metabolism level may predict a fatal outcome after H7N9 virus infection.Copyright © 2018. Published by Elsevier B.V.

Keyword: inflammation

is a novel hypothalamic gene upregulated by a high-fat diet and leptin in mice.

Energy homeostasis is regulated by the hypothalamus but fails when animals are fed a high-fat diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated fat, oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic .

Keyword: inflammation

Macrophage-Specific Hypoxia-Inducible Factor-1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis.

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia-inducible factors (HIFs) have recently been identified as important regulators of immunity and . In nonalcoholic steatohepatitis (NASH), HIF-1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF-1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF-1α levels in hepatic macrophages in methionine-choline-deficient (MCD) diet-fed mice and in macrophages of patients with NASH compared with controls. The HIF-1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin-1, LC3-II, and p62 in both mouse and human macrophages. LysM HIF fl/fl mice, which have HIF-1α levels stabilized in macrophages, showed higher steatosis and liver compared with HIF fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated fatty , (PA), both induces HIF-1α and impairs autophagic flux in macrophages. Using small interfering RNA-mediated knock-down and overexpression of HIF-1α in macrophages, we demonstrated that PA impairs autophagy via HIF-1α. We found that HIF-1α mediates NF-κB activation and MCP-1 production and that HIF-1α-mediated impairment of macrophage autophagy increases IL-1β production, contributing to MCD diet-induced NASH. Conclusion: impairs autophagy via HIF-1α activation in macrophages. HIF-1α and impaired autophagy are present in NASH in vivo in mouse macrophages and in human blood monocytes. We identified that HIF-1α activation and decreased autophagic flux stimulate in macrophages through upregulation of NF-κB activation. These results suggest that macrophage activation in NASH involves a complex interplay between HIF-1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet-induced NASH.© 2018 by the American Association for the Study of Liver Diseases.

Keyword: inflammation

Modification of gene expression in rat cardiomyocytes by linoleic and docosahexaenoic acids .

Regulation of cardiac fatty metabolism is central to the development of cardiac hypertrophy and heart failure. We investigated the effects of select fatty acids on the expression of genes involved in immediate early as well as inflammatory and hypertrophic responses in adult rat cardiomyocytes. Cardiac remodeling begins with upregulation of immediate early genes for c-fos and c-jun, followed by upregulation of inflammatory genes for nuclear factor kappa B (NF-κB) and nuclear factor of activated T-cells (NFAT). At later stages, genes involved in hypertrophic responses, such as atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), are upregulated. Adult rat cardiomyocytes were treated with , a saturated fatty ; oleic , a monounsaturated fatty ; linoleic , a polyunsaturated fatty belonging to the n-6 class; and docosahexaenoic , a polyunsaturated fatty belonging to the n-3 class. Linoleic produced a greater increase in the mRNA expression of c-fos, c-jun, NF-κB, NFAT3, ANP, and BNP relative to and oleic . In contrast, docosahexaenoic caused a decrease in the expression of genes involved in cardiac hypertrophy. Our findings suggest that linoleic may be a potent inducer of genes involved in cardiac hypertrophy, whereas docosahexaenoic may be protective against the cardiomyocyte hypertrophic response.

Keyword: inflammation

Acetone fraction from Sechium edule (Jacq.) S.w. edible roots exhibits anti-endothelial dysfunction activity.

A recent ethnomedical survey on medicinal plants grown in Mexico revealed that Sechium edule (Jacq.) Sw. (Cucurbitaceae) is one of the most valued plant species to treat cardiovascular diseases, including hypertension. Fruits, young leaves, buds, stems, and tuberous roots of the plant are edible. Considering that endothelial dysfunction induced by Angiotensin II plays an important role in the pathogenesis of hypertension and is accompanied by a prooxidative condition, which in turn induces an inflammatory state, vascular remodeling, and tissue damage, and that S. edule has been reported to possess antioxidant, anti-inflammatory and antihypertensive activity, its capability to control endothelial dysfunction was also assessed.To assess in vivo the anti-endothelial dysfunction activity of the acetone fraction (rSe-ACE) of the hydroalcoholic extract from S. edule roots.Endothelial dysfunction was induced in female C57BL/6\u202fJ mice by a daily intraperitoneal injection of angiotensin II for 10 weeks. Either rSe-ACE or losartan (as a control) were co-administered with angiotensin II for the same period. Blood pressure was measured at weeks 0, 5, and 10. Kidney extracts were prepared to determine IL1β, IL4, IL6, IL10, IL17, IFNγ, TNFα, and TGFβ levels by ELISA, along with the prooxidative status as assessed by the activity of antioxidant enzymes. The expression of ICAM-1 was evaluated by immunohistochemistry in kidney histological sections. Kidney and hepatic damage, as well as vascular tissue remodeling, were studied.The rSe-ACE fraction administered at a dose of 10\u202fmg/kg was able to control hypertension, as well as the prooxidative and proinflammatory status in kidney as efficiently as losartan, returning mice to normotensive levels. Additionally, the fraction was more efficient than losartan to prevent liver and kidney damage. Phytochemical characterization identified cinnamic as a major compound, and linoleic, , and myristic acids as the most abundant non-polar components in the mixture, previously reported to aid in the control of hypertension, , and oxidative stress, three important components of endothelial dysfunction.this study demonstrated that rSe-ACE has anti-endothelial dysfunction activity in an experimental model and highlights the role of cinnamic and fatty acids in the observed effects.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, , and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free fatty (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: inflammation

damages gut epithelium and initiates inflammatory cytokine production.

The mechanisms leading to the low-grade inflammation observed during obesity are not fully understood. Seeking the initiating events, we tested the hypothesis that the intestine could be damaged by repeated lipid supply and therefore participate in inflammation. In mice, 1-5 palm oil gavages increased intestinal permeability via decreased expression and mislocalization of junctional proteins at the cell-cell contacts; altered the intestinal bacterial species by decreasing the abundance of Akkermansia muciniphila, segmented filamentous bacteria, and Clostridium leptum; and increased inflammatory cytokine expression. This was further studied in human intestinal epithelial Caco-2/TC7 cells using the two main components of palm oil, i.e., and oleic . Saturated impaired paracellular permeability and junctional protein localization, and induced inflammatory cytokine expression in the cells, but unsaturated oleic did not. Inhibiting de novo ceramide synthesis prevented part of these effects. Altogether, our data show that short exposure to palm oil or induces intestinal dysfunctions targeting and inflammation. Excessive palm oil consumption could be an early player in the gut alterations observed in metabolic diseases.Copyright © 2019. Published by Elsevier B.V.

Keyword: inflammation

Romidepsin suppresses monosodium urate crystal-induced cytokine production through upregulation of suppressor of cytokine signaling 1 expression.

Acute gouty arthritis currently is the most common form of inflammatory arthritis in developed countries. Treatment is still suboptimal. Dosage of urate-lowering therapy is often too low to reach target urate levels, and adherence to therapy is poor. In this study, we therefore explore a new treatment option to limit in acute gout: specific histone deacetylase (HDAC) inhibition.Peripheral blood mononuclear cells (PBMCs) were cultured with a combination of monosodium urate crystals (MSU) and (C16.0) in order to activate the NLRP3 inflammasome and induce IL-1β production. HDAC inhibitors and other compounds were added beforehand with a 1-h pre-incubation period.The HDAC1/2 inhibitor romidepsin was most potent in lowering C16.0+MSU-induced IL-1β production compared to other specific class I HDAC inhibitors. At 10\u2009nM, romidepsin decreased IL-1β, IL-1Ra, IL-6, and IL-8 production. IL-1β mRNA was significantly decreased at 25\u2009nM. Although romidepsin increased PTEN expression, PBMCs from patients with germline mutations in PTEN still responded well to romidepsin. Romidepsin also increased SOCS1 expression and blocked STAT1 and STAT3 activation. Furthermore, experiments with bortezomib showed that blocking the proteasome reverses the cytokine suppression by romidepsin.Our results show that romidepsin is a very potent inhibitor of C16.0+MSU-induced cytokines in vitro. Romidepsin upregulated transcription of SOCS1, which was shown to directly target inflammatory signaling molecules for proteasomal degradation. Inhibiting the proteasome therefore reversed the cytokine-suppressive effects of romidepsin. HDAC1/2 dual inhibition could therefore be a highly potent new treatment option for acute gout, although safety has to be determined in vivo.

Keyword: inflammation

Integrated metabolomics and network toxicology to reveal molecular mechanism of celastrol induced cardiotoxicity.

Celastrol (CS), an active triterpene derived from traditional Chinese medicine Tripterygium wilfordii Hook. f, has been used to treat chronic inflammation, arthritis and other diseases. However, it has been reported that CS can trigger cardiotoxicity and the molecular mechanism of heart injury induced by CS is not clear. Considering the wide application of Tripterygium wilfordii Hook. f in clinics, it is necessary to develop an accurate and reliable method to assess the safety of CS, and to elucidate as much as possible the mechanism of cardiotoxicity induced by CS. In this study, Ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics revealed clues to the mechanism of CS-induced heart injury. significantly increased in plasma from CS-treated rats, and this increase resulted in oxidative stress response in vivo. Excessive ROS further activate TNF signaling pathway and caspase family, which were obtained from the KEGG enrichment analysis of network toxicology strategy. Protein expression level of caspase-3, caspase-8, bax were significantly increased by western blot. Q-PCR also showed the similar results as western blot. It means that apoptosis plays a key role in the process of celastrol induced cardiotoxicity. Blocking this signal axis may be a potential way to protect myocardial tissue.Copyright © 2019. Published by Elsevier Inc.

Keyword: inflammation

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial lipid accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic lipid metabolism. Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 diabetes animal models. After streptozotocin (STZ) or high-fat diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, lipid accumulation, oxidative stress, and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and lipid accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing lipid accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Obesity-related cellular stressors regulate gonadotropin releasing hormone gene expression via c-Fos/AP-1.

Obesity is a risk factor for infertility, but mechanisms underlying this risk are unclear. Fertility is regulated by hypothalamic gonadotropin-releasing hormone, encoded by the Gnrh1 gene. Because obesity promotes endoplasmic reticulum (ER) stress, we sought to determine how tunicamycin-induced ER stress affected Gnrh1 gene expression in the mouse hypothalamic cell line GT1-7. Tunicamycin repressed expression of Gnrh1 in a PKC- and JNK-dependent manner, while upregulating expression of a known Gnrh1 repressor, Fos. Obesity is associated with increased circulating free fatty acids, and exposure to palmitate promoted ER stress and . Fos expression increased with palmitate dose, but Gnrh1 expression was upregulated with low-dose palmitate and repressed with high-dose palmitate. Using a small molecule inhibitor, we determined that AP-1 was required for Gnrh1 repression by high-dose palmitate or tunicamycin-induced ER stress. These findings suggest that hypogonadism driven by decreased hypothalamic GnRH may be a component of obesity-related infertility.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

MicroRNA-155 Mediates Obesity-Induced Renal and Dysfunction.

Chronic is a major contributor to obesity-related renal damage. Recent studies have demonstrated that microRNA (miR)-155 is closely associated with hyperglycemia-induced nephropathy, but whether renal miR-155 participates in the inflammatory response and development of obesity-related nephropathy is unknown. In present study, we investigated the pathophysiological role of renal miR-155 in (PA)-treated endothelial cell and high-fat-diet (HFD)-fed mouse models by specific miR-155 sponge. Mice fed with HFD exhibited higher levels of renal miR-155, which positively correlated with urine microalbumin and blood urea nitrogen. In vitro study, mouse renal vascular endothelial cells stimulated with PA also showed higher miR-155 levels, accompanied with increased inflammatory response. Suppression of renal miR-155 effectively attenuated HFD-induced renal structural damages and dysfunction. MiR-155 sponge treatment also significantly decreased NF-κB signaling and downstream gene expression in vitro and in vivo. The obesity-increased macrophage infiltration and lipotoxicity was decreased in mouse kidney after miR-155 sponge treatment. Mechanistically, miR-155 directly targeted 3\'-UTR of SHIP1/INPP5D and suppressed its expression in vitro and in vivo, whereas silence of SHIP1/INPP5D abolished the renal protective benefits of miR-155 sponge in obese mice. Taken together, present findings for the first time provided evidence for the potential role of miR-155 in obesity-related nephropathy and clarified that SHIP1/NF-κB signaling was a potential molecular mechanism.

Keyword: inflammation

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid Metabolism, Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free fatty (FFA)- and oleic -and high-fat diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice liver were measured, and the expression levels of genes associated with lipogenesis, fatty oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with fatty oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of lipid metabolism, inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: inflammation

Alkannin Inhibited Hepatic in Diabetic Db/Db Mice.

The current study was designed to investigate the protective role of alkannin (ALK) on liver injury in diabetic C57BL/KsJ-db/db mice and explore its potential mechanisms.An oral glucose tolerance test (OGTT) was performed. The levels of insulin, alanine aminotransferase (ALT), aspartate aminotransaminase (AST), total cholesterol (TC) and triglyceride (TG) were determined by commercial kits. The pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were determined by ELISA. The levels of the ROCK/NF-κB pathway were determined by Western blotting.The contents of pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were inhibited by ALK, metformin or fasudil in diabetic db/db mice. Further, Western blotting analysis showed that the expression of Rho, ROCK1, ROCK2, p-NF-κBp65, and p-IκBα was significantly reversed by ALK treatment. In human hepatic HepG2 cells, the hepatoprotective effects of ALK were further characterized. With response to -challenge, increased amounts of insulin, ALT, AST, TG, and TC were observed, whereas ALK pretreatment significantly inhibited their leakage in HepG2 cells without appreciable cytotoxic effects. The condition was recovered with ALK treatment as shown by changes of IL-1β, IL-6 and TNF-α. Further, Western blotting analysis also suggested that ALK improves hepatic in a Rho-kinase pathway.The present study successfully investigated the role of Rho-kinase signalling in diabetic liver injury. ALK exhibited hepatoprotective effects in diabetic db/db mice, and it might act through improving hepatic through the Rho-kinase pathway.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: inflammation

Treatment of cigarette smoke extract and condensate differentially potentiates -induced lipotoxicity and steatohepatitis in vitro.

Accumulative evidence showed that cigarette smoke (CS) detrimentally affects the pathogenesis of nonalcoholic steatohepatitis (NASH). The purpose of this study was to evaluate the effects of CS extract (CSE) or total particulate matter (TPM) on the in vitro steatohepatitis model using mouse primary hepatocytes treated with (PA) or PA plus LPS. Increased hepatocellular damage was observed in PA-treated hepatocytes with TPM or CSE treatment, but increased triglyceride level was only observed in PA plus LPS-treated hepatocytes with a high concentration of TPM. Also, expression levels of steatohepatitis-related genes such as TNF-α, NOS 2, and SREBP-1c were significantly increased after treatment of TPM. To further demonstrate the role of Kupffer cells (KCs) after CS extracts treatment, trans-well co-culture system of hepatocytes and KCs was utilized. The levels of inflammatory cytokines and the ratios of Bax/Bcl-2 (apoptosis-related genes) were markedly increased in co-cultured hepatocytes after TPM or CSE treatment. Interestingly, KCs activation was augmented in KCs upon treatment with CSE or TPM. Overall, our findings indicate that in vitro treatment with CSE or TPM differentially contributes to the severity of steatohepatitis by modulating steatohepatitis-related lipotoxicity and , which might be caused by KCs activation with subsequent induction of hepatocytes apoptosis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Astragalosides IV protected the renal tubular epithelial cells from free fatty acids-induced injury by reducing oxidative stress and apoptosis.

Renal tubular injury is associated with the development of diabetic nephropathy (DN) and the end-stage renal disease (ESRD). Free fatty acids (FFAs)-associated lipotoxicity contributes to injury of proximal renal tubular epithelial (HK-2) cells in diabetes. (PA) which is the most abundant saturated fatty in FFAs is closely associated with the gradual decline of renal function. Astragalosides IV (AS-IV) has a variety of pharmacological effects such as anti- and anti-oxidation. In the current study, we investigated the effects of AS-IV on PA-induced apoptosis of HK-2 cells and the underlying mechanisms. The results showed that AS-IV (10, 20, 40\u2009μmol/L) could alleviate PA-induced apoptosis of HK-2 cells. We found that AS-IV reduced the expression of Bax and cleaved-caspase3, but increased the expression of Bcl-2 and phosphorylated Nrf2 in HK-2 cells. Moreover, AS-IV reduced the level of reactive oxygen species (ROS) in the cells. Our study suggests that AS-IV could protect against PA-induced apoptosis in HK-2 cells by inhibiting ROS generation and apoptotic protein expression. This study may provide a new theoretical option for the patients with type 2 diabetes.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: inflammation

The effect of on inflammatory response in macrophages: an overview of molecular mechanisms.

is a saturated fatty whose blood concentration is elevated in obese patients. This causes inflammatory responses, where toll-like receptors (TLR), TLR2 and TLR4, play an important role. Nevertheless, is not only a TLR agonist. In the cell, this fatty is converted into phospholipids, diacylglycerol and ceramides. They trigger the activation of various signaling pathways that are common for LPS-mediated TLR4 activation. In particular, metabolic products of affect the activation of various PKCs, ER stress and cause an increase in ROS generation. Thanks to this, also strengthens the TLR4-induced signaling. In this review, we discuss the mechanisms of inflammatory response induced by . In particular, we focus on describing its effect on ER stress and IRE1α, and the mechanisms of NF-κB activation. We also present the mechanisms of inflammasome NLRP3 activation and the effect of on enhanced inflammatory response by increasing the expression of FABP4/aP2. Finally, we focus on the consequences of inflammatory responses, in particular, the effect of TNF-α, IL-1β and IL-6 on insulin resistance. Due to the high importance of macrophages and the production of proinflammatory cytokines by them, this work mainly focuses on these cells.

Keyword: inflammation

Cathepsin B inhibition ameliorates the non-alcoholic steatohepatitis through suppressing caspase-1 activation.

Non-alcoholic fatty liver disease (NAFLD) has emerged as the most common chronic liver disease. NLRP3 inflammasome activation has been widely studied in the pathogenesis of NAFLD. Cathepsin B (CTSB) is a ubiquitous cysteine cathepsin, and the role of CTSB in the progression and development of NAFLD has received extensive concern. However, the exact roles of CTSB in the NAFLD development and NLRP3 inflammasome activation are yet to be evaluated. In the present study, we used methionine choline-deficient (MCD) diet to establish mice NASH model. CTSB inhibitor (CA-074) was used to suppress the expression of CSTB. Expressions of CTSB and caspase-1 were evaluated by immunohistochemical staining. Serum IL-1β and IL-18 levels were also determined. was used to stimulate Kupffer cells (KCs), and protein expressions of CTSB, NLRP3, ASC (apoptosis-associated speck-like protein containing CARD), and caspase-1 in KCs were detected. The levels of IL-1β and IL-18 in the supernatant of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). Our results showed that CTSB inhibition improved the liver function and reduced hepatic and ballooning, and the levels of pro-inflammatory cytokines IL-1β and IL-18 were decreased. The expressions of CTSB and caspase-1 in liver tissues were increased in the NASH group. In in vitro experiments, PA stimulation could increase the expressions of CTSB and NLRP3 inflammasome in KCs, and CTSB inhibition downregulated the expression of NLRP3 inflammasome in KCs, when challenged by PA. Moreover, CTSB inhibition effectively suppressed the expression and activity of caspase-1 and subsequently secretions of IL-1β and IL-18. Collectively, these results suggest that CTSB inhibition limits NLRP3 inflammasome-dependent NASH formation through regulating the expression and activity of caspase-1, thus providing a novel anti-inflammatory signal pathway for the therapy of NAFLD.

Keyword: inflammation

Degradation of splicing factor SRSF3 contributes to progressive liver disease.

Serine rich splicing factor 3 (SRSF3) plays a critical role in liver function and its loss promotes chronic liver damage and regeneration. As a consequence, genetic deletion of SRSF3 in hepatocytes caused progressive liver disease and ultimately led to hepatocellular carcinoma. Here we show that SRSF3 is decreased in human liver samples with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or cirrhosis that was associated with alterations in RNA splicing of known SRSF3 target genes. Hepatic SRSF3 expression was similarly decreased and RNA splicing dysregulated in mouse models of NAFLD and NASH. We showed that -induced oxidative stress caused conjugation of the ubiquitin like NEDD8 protein to SRSF3 and proteasome mediated degradation. SRSF3 was selectively neddylated at lysine11 and mutation of this residue (SRSF3-K11R) was sufficient to prevent both SRSF3 degradation and alterations in RNA splicing. Finally prevention of SRSF3 degradation in vivo partially protected mice from hepatic steatosis, fibrosis and . These results highlight a neddylation-dependent mechanism regulating gene expression in the liver that is disrupted in early metabolic liver disease and may contribute to the progression to NASH, cirrhosis and ultimately hepatocellular carcinoma.

Keyword: inflammation

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid metabolism that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty metabolism and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty metabolism. Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: inflammation

Regulatory roles of miR-155 and let-7b on the expression of -related genes in THP-1 cells: effects of fatty acids.

The main aim of this investigation was to study the regulatory roles of let-7b and miR-155-3p on the expression of -associated genes in monocytes, macrophages, and lipopolysaccharide (LPS)-activated macrophages (AcM). A second goal was to analyze the potential modulatory roles of different fatty acids, including oleic, , eicosapentaenoic (EPA), and docosahexaenoic (DHA), on the expression of these miRNAs in the three cell types. This hypothesis was tested in human acute monocytic leukemia cells (THP-1), which were differentiated into macrophages with 2-O-tetradecanoylphorbol-13-acetate (TPA) and further activated with LPS for 24\xa0h. Monocytes, macrophages, and AcM were transfected with a negative control, or mimics for miR-155-3p and miR-let-7b-5p. The expression of both miRNAs and some proinflammatory genes was analyzed by qRT-PCR. Interestingly, let-7b mimic reduced the expression of IL6 and TNF in monocytes, and SERPINE1 expression in LPS-activated macrophages. However, IL6, TNF, and SERPINE1 were upregulated in macrophages by let-7b mimic. IL6 expression was higher in the three types of cells after transfecting with miR-155-3p mimic. Similarly, expression of SERPINE1 was increased by miR-155-3p mimic in monocytes and macrophages. However, TLR4 was downregulated by miR-155-3p in monocytes and macrophages. Regarding the effects of the different fatty acids, oleic increased the expression of let-7b in macrophages and AcM and also increased the expression of miR-155 in monocytes when compared with DHA but not when compared with non-treated cells. Overall, these results suggest anti- and proinflammatory roles of let-7b and miR-155-3p in THP-1 cells, respectively, although these outcomes are strongly dependent on the cell type. Noteworthy, oleic might exert beneficial anti-inflammatory effects in immune cells (i.e., non-activated and LPS-activated macrophages) by upregulating the expression of let-7b.

Keyword: inflammation

Role of the mTOR‑FOXO1 pathway in obesity‑associated renal tubulointerstitial .

Since obesity is largely responsible for the growing incidence of renal tubulointerstitial , exploration into the mechanisms of obesity‑associated tubulointerstitial is essential. Studies have demonstrated that mammalian target of rapamycin\xa0(mTOR) is a crucial molecule in the pathogenesis of renal , including regulating the expression of inflammatory factors. The purpose of the present study was to further elucidate the role of mTOR in obesity‑associated tubulointerstitial . In the clinical study, obese and healthy subjects were recruited for physical examination, as well as the collection of blood and urine samples. Further study was performed on a high fat diet\xa0(HFD)‑induced obese rat model and a cultured human renal tubular epithelial cell line\xa0(HK‑2). The clinical study demonstrated that the participants with obesity had increased serum lipids, creatinine\xa0(Cr), urinary albumin to creatinine ratio\xa0(UACR) and urinary neutrophil gelatinase‑associated lipocalin\xa0(u‑NGAL). Moreover, the level of urinary monocyte chemoattractant protein‑1\xa0(u‑MCP‑1) was increased in the participants with obesity, and it was positively correlated with free fatty \xa0(FFA), UACR and u‑NGAL. In the in\xa0vivo study, the results indicated that the levels of serum lipids, Cr and blood urea nitrogen\xa0(BUN), as well as 24\xa0h urine protein and u‑NGAL, were significantly increased in the HFD‑fed obese rats. In addition, the infiltration of CD68+ cells into the renal interstitial area and the release of interleukin‑1β\xa0(IL‑1β) was observed in the kidneys of obese rats. Meanwhile, the supernatant from HK‑2 cells treated with stimulated THP‑1 monocyte migration. The upregulation of MCP‑1, phosphorylated forkhead boxO1 (p‑FOXO1), and phosphorylated mTOR (p‑mTOR) was observed in\xa0vivo and in\xa0vitro. However, inhibition of mTOR was able to alleviate the above effects. Overall, these results demonstrated that activated mTOR induced FOXO1 phosphorylation, which mediates renal MCP‑1 release, causes tubulointerstitial and ultimately leads to pathological renal changes and dysfunction. However, inhibition of mTOR may play a renoprotective role during the progression of obesity‑associated tubulointerstitial .

Keyword: inflammation

Complement Component C3 Is Highly Expressed in Human Pancreatic Islets and Prevents β Cell Death via ATG16L1 Interaction and Autophagy Regulation.

We show here that human pancreatic islets highly express C3, which is both secreted and present in the cytosol. Within isolated human islets, C3 expression correlates with type 2 diabetes (T2D) donor status, HbA1c, and . Islet C3 expression is also upregulated in several rodent diabetes models. C3 interacts with ATG16L1, which is essential for autophagy. Autophagy relieves cellular stresses faced by β cells during T2D and maintains cellular homeostasis. C3 knockout in clonal β cells impaired autophagy and led to increased apoptosis after exposure of cells to and IAPP. In the absence of C3, autophagosomes do not undergo fusion with lysosomes. Thus, C3 may be upregulated in islets during T2D as a cytoprotective factor against β cell dysfunction caused by impaired autophagy. Therefore, we revealed a previously undescribed intracellular function for C3, connecting the complement system directly to autophagy, with a broad\xa0potential importance in other diseases and cell types.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Macrophages with a deletion of the () gene have a more proinflammatory phenotype.

Phosphoenolpyruvate carboxykinase (Pck1) is a metabolic enzyme that is integral to the gluconeogenic and glyceroneogenic pathways. However, Pck1\'s role in macrophage metabolism and function is unknown. Using stable isotopomer MS analysis in a mouse model with a myeloid cell-specific deletion, we show here that this deletion increases the proinflammatory phenotype in macrophages. Incubation of LPS-stimulated bone marrow-derived macrophages (BMDM) with [U-C]glucose revealed reduced C labeling of citrate and malate and increased C labeling of lactate in Pck1-deleted bone marrow-derived macrophages. We also found that the Pck1 deletion in the myeloid cells increases reactive oxygen species (ROS). Of note, this altered macrophage metabolism increased expression of the M1 cytokines TNFα, IL-1β, and IL-6. We therefore conclude that contributes to M1 polarization in macrophages. Our findings provide important insights into the factors determining the macrophage inflammatory response and indicate that Pck1 activity contributes to metabolic reprogramming and polarization in macrophages.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: inflammation

Emex spinosa (L.) Campd. ethyl acetate fractions effects on and oxidative stress markers in carrageenan induced paw oedema in mice.

Emex spinosa (L.) Campd. (E. spinosa) locally known as "hillaioua" has always been used in folk medicine for the treatment of and pain. It is still being exploited by pharmaceutical companies for its potential remedial effects.In this study, the effects of E. spinosa (L.) Campd. against acute , pain and oxidative damage were evaluated.Total phenols and flavonoids were evaluated. Anti-inflammatory and analgesic activities the E. spinosa ethyl acetate fractions of the aerial (Es EtOAc-AP) and underground (Es EtOAc-R) parts were assessed on carrageenan-induced paw oedema (100\u202fmg/kg BW) and acetic -induced writhing response (50, 100 and 150\u202fmg/kg BW), respectively. The E. spinosa fractions effects on oxidative stress markers and inflammatory parameters were determined. Gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify various chemical components.The ethyl acetate fractions were shown to be the most active thanks to their phenolic and flavonoid contents richness. Intraperitoneal administration of E. spinosa ethyl acetate fractions at 100\u202fmg/kg BW, one hour before carrageenan injection, significantly inhibited the oedema formation by 89.31% and 97.7% for the aerial and underground parts respectively when compared to the reference drug "dexamethasone"\u202f(51.9%). Besides, a significant increase (p\u202f≤\u202f0.001) of the dermal antioxidant enzymes (the superoxide dismutase (SOD)), catalase (CAT) and glutathione peroxidase (GPx) was observed five hours after carrageenan administration. The best restoration was obtained with Es EtOAc-R (82.04%, 93.55% and 93.55% respectively for SOD, CAT and GPx activities). Moreover, EtOAc-fractions treated mice proved their ability to restore both of CRP and fibrinogen (p\u202f<\u202f0.001). In addition, E. spinosa EtOAc-fractions attenuated abdominal contractions (p\u202f<\u202f0.05) by 71.69% and 82.41% for the aerial part and roots respectively at 150\u202fmg/kg BW against 100% for dichlofenac sodium used as standard drug. The phytochemical analysis of Es EtOAc-AP and Es EtOAc-R by GC-MS may explain the obtained results. The analysis of the fractions demonstrated the presence of and linoleic acids known for their anti-inflammatory and analgesic capacities.These findings explain the traditional use of E. spinosa in folk medicine and suggest that E. spinosa fractions could be a promising herbal drug.Copyright © 2018. Published by Elsevier B.V.

Keyword: inflammation

Comparison of diets enriched in stearic, oleic, and acids on , immune response, cardiometabolic risk factors, and fecal bile concentrations in mildly hypercholesterolemic postmenopausal women-randomized crossover trial.

Direct comparisons between SFAs varying in chain length, specifically (16:0) and stearic (18:0), relative to the latter\'s metabolic product, oleic (18:1), on cardiometabolic risk factors are limited.The aim of this study was to determine the relative comparability of diets enriched in , stearic , and oleic on and coagulation markers, T lymphocyte proliferation/ex-vivo cytokine secretion, plasma cardiometabolic risk factors, and fecal bile concentrations.Hypercholesterolemic postmenopausal women (n\xa0=\xa020, mean\xa0±\xa0SD age 64\xa0±\xa07 y, BMI 26.4\xa0±\xa03.4 kg/m2, LDL cholesterol\xa0≥\xa02.8 mmol/L) were provided with each of 3 diets [55% energy (%E) carbohydrate, 15%E protein, 30%E fat, with ∼50% fat contributed by , stearic , or oleic in each diet; 5 wk/diet phase] using a randomized crossover design with 2-wk washouts between phases. Outcome measures were assessed at the end of each phase.Fasting LDL-cholesterol and non-HDL-cholesterol concentrations were lower after the stearic and oleic diets than the diet (all P\xa0<\xa00.01). Fasting HDL-cholesterol concentrations were lower after the stearic diet than the and oleic diets (P\xa0<\xa00.01). The stearic diet resulted in lower lithocholic (P\xa0=\xa00.01) and total secondary bile (SBA) concentrations (P\xa0=\xa00.04) than the oleic diet. All other outcome measures were similar between diets. Lithocholic concentrations were positively correlated with fasting LDL-cholesterol concentrations (r\xa0=\xa00.33; P\xa0=\xa00.011). Total SBA, lithocholic , and deoxycholic concentrations were negatively correlated with fasting HDL cholesterol (r\xa0=\xa0-0.51 to -0.44; P\xa0<\xa00.01) concentrations and positively correlated with LDL cholesterol:HDL cholesterol (r =\xa00.37-0.54; P\xa0<\xa00.01) ratios.Dietary stearic and oleic had similar effects on fasting LDL-cholesterol and non-HDL-cholesterol concentrations and more favorable ones than . Unlike oleic , the hypocholesterolemic effect of stearic may be mediated by inhibition of intestinal hydrophobic SBA synthesis. These findings add to the data suggesting there should be a reassessment of current SFA dietary guidance and Nutrient Facts panel labeling.This trial was registered at clinicaltrials.gov as .Copyright © American Society for Nutrition 2019.

Keyword: inflammation

Plasma Saturated and Monounsaturated Fatty Acids in Behçet\'s Disease.

Fatty (FA) composition of serum has been associated with many markers of . In this study, we tried to examine plasma Saturated Fatty (SFA) and Monounsaturated Fatty (MUFA) composition in Behçet\'s Disease (BD) patients. The associations between the circulating FA levels and some markers of have also been investigated.This study is a cross-sectional one. In fact, a total of 101 BD patients and healthy controls group of 99 subjects are enrolled. Gas Chromatograph equipped with a Capillary Split/Splitless Injector and flame ionization detector was used to analyze the plasma SFA and MUFA compositions. The high sensitivity C-Reactive Protein (hsCRP) and fibrinogen levels were measured using standard techniques.BD patients had significantly higher proportions of Mystiric (MA), (PAM), Palmitoleic (POA) and Stearoyl-CoA Desaturase (SCD)-16, compared to controls.The results revealed that patients with severe involvements had high levels of POA and total MUFA associated with higher SCD-16 activity compared to those with minor ones. The receiver operator characteristic curve analysis revealed that POA could well discriminate BD patients with severe clinical manifestations. In the bivariate analysis, hsCRP was found to be positively correlated with total SAFA and POA elongase activity index but negatively correlated with SCD-18 activity index. The STA, POA, elongase and SCD-16 activity index are correlated with fibrinogen. On the other hand, the multivariate analysis showed that POA remained associated with higher levels of hsCRP.Unfavourable plasma SFA and MUFA profile were reported in BD patients. POA, which is associated with higher plasma hsCRP level, may play a role in the pathogenesis of BD.

Keyword: inflammation

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe liver and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: inflammation

Inhibition of protein kinase R protects against -induced , oxidative stress, and apoptosis through the JNK/NF-kB/NLRP3 pathway in cultured H9C2 cardiomyocytes.

Double-stranded RNA-dependent protein kinase (PKR) is a critical regulator of apoptosis, oxidative stress, and under hyperlipidemic and insulin resistance conditions. Saturated free fatty acids, such as (PA), are known inducers of apoptosis in numerous cell types. However, the underlying molecular mechanism is not fully understood. The aim of the present study was to examine the effect of PA on cultured rat H9C2 cardiac myocytes cells and to investigate the PKR mediated harmful effects of PA in vitro in cultured cardiomyocytes.PKR expression was determined by immunofluorescence and immunoblotting. Oxidative stress and apoptosis were determined by flow cytometry and assay kits. The expression of different gene markers of apoptosis, oxidative stress, and were measured by Western blot analysis and reverse transcription polymerase chain reaction.PKR expression, reactive oxygen species levels as well as apoptosis were increased in PA-treated cultured H9C2 cardiomyocytes. The harmful effects of PA were attenuated by a selective PKR inhibitor, C16. Moreover, we observed that upregulation of c-Jun N-terminal kinase (JNK), nuclear factor-kB (NF-kB) and NACHT, LRR and PYD domains-containing protein 3 (NLRP3) pathways is associated with increased expression of interleukin 6 and tumor necrosis factor-α in PA-treated cardiomyocytes and attenuation by a selective PKR inhibitor.Our study reports, for the first time, that PKR-mediated harmful effects of PA in cultured cardiomyocytes via activation of JNK, NF-kB, and NLRP3 pathways. Inhibition of PKR is one of the possible mechanistic approaches to inhibit , oxidative stress, and apoptosis in lipotoxicity-induced cardiomyocyte damage.© 2018 Wiley Periodicals, Inc.

Keyword: inflammation

Vaginal lipidomics of women with vulvovaginal candidiasis and cytolytic vaginosis: A non-targeted LC-MS pilot study.

To characterize the lipid profile in vaginal discharge of women with vulvovaginal candidiasis, cytolytic vaginosis, or no vaginal infection or dysbiosis.Cross-sectional study.Genital Infections Ambulatory, Department of Tocogynecology, University of Campinas, Campinas, São Paulo-Brazil.Twenty-four women were included in this study: eight with vulvovaginal candidiasis, eight with cytolytic vaginosis and eight with no vaginal infections or dysbiosis (control group).The lipid profile in vaginal discharge of the different study groups was determined by liquid chromatography-mass spectrometry and further analyzed with MetaboAnalyst 3.0 platform.Vaginal lipids concentration and its correlation with vulvovaginal candidiasis and cytolytic vaginosis.PCA, PLS-DA and hierarchical clustering analyses indicated 38 potential lipid biomarkers for the different groups, correlating with oxidative stress, , apoptosis and integrity of the vaginal epithelial tissue. Among these, greater concentrations were found for Glycochenodeoxycholic -7-sulfate, O-adipoylcarnitine, 1-eicosyl-2-heptadecanoyl-glycero-3-phosphoserine, undecanoic , formyl dodecanoate and lipoic in the vulvovaginal candidiasis group; N-(tetradecanoyl)-sphinganine, DL-PPMP, 1-oleoyl-cyclic phosphatidic, and 5-aminopentanoic in the cytolytic vaginosis group; and 1-nonadecanoyl-glycero-3-phosphate, eicosadienoic , 1-stearoyl-cyclic-phosphatidic , 1-(9Z,12Z-heptadecadienoyl)-glycero-3-phosphate, formyl 9Z-tetradecenoate and 7Z,10Z-hexadecadienoic in the control group.Lipids related to oxidative stress and apoptosis were found in higher concentrations in women with vulvovaginal candidiasis and cytolytic vaginosis, while lipids related to epithelial tissue integrity were more pronounced in the control group. Furthermore, in women with cytolytic vaginosis, we observed higher concentrations of lipids related to bacterial overgrowth.

Keyword: inflammation

Growth, stool consistency and bone mineral content in healthy term infants fed sn-2-palmitate-enriched starter infant formula: A randomized, double-blind, multicentre clinical trial.

Palmitate in breast milk is predominantly located in the triacylglycerol sn-2 position, while infant formulae contain palmitate predominantly in the sn-1 and sn-3 positions. During digestion, palmitate in the sn-1 and sn-3 positions is hydrolyzed to free that can subsequently complex with calcium to form insoluble soaps; this may partially explain why formula-fed infants have harder stools than breast-fed infants.This large (n\xa0=\xa0488) randomized, double-blind, multicentre trial investigated whether increasing the sn-2 palmitate content of infant formula improves stool consistency and bone mineral content (measured by dual-energy x-ray absorptiometry), without affecting growth or health. From ∼1 week to 4 months of age, infants were exclusively fed one of three formulae: i) control formula (CF; 16% of total palmitate at sn-2; n\xa0=\xa0162), (ii) experimental formula 1 (EF1; 43% of total palmitate at sn-2; n\xa0=\xa0166) or (iii) experimental formula 2 (EF2; 51% of total palmitate at sn-2; n\xa0=\xa0160).Intention-to-treat analysis showed softer stools in both EF groups (vs. CF) at ages 2 weeks and 1 and 2 months (p\xa0≤\xa00.01), but not 3 and 4 months. At 4 months, all groups had similar growth outcomes while bone mineral content was significantly higher in EF1 (p\xa0=\xa00.0012) and EF2 (p\xa0=\xa00.0002) compared with CF. Comparison of reported adverse events up to 12 months revealed no differences among groups. All 3 infant formulae exhibited equally good digestive tolerance.Formulae enriched in sn-2 palmitate fed in early infancy are safe, improve stool consistency (from 2 weeks to 2 months) and increase bone mineral content (at 4 months).Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Keyword: inflammation

Saturated and unsaturated fatty acids differentially regulate in vitro and ex vivo placental antioxidant capacity.

Complications from prematurity are the leading cause of death among children under 5\xa0years of age. Although clinical studies have shown a positive correlation between maternal high-fat diet (HFD) and preterm birth (PTB), the underlying mechanisms remain to be elucidated. Furthermore, it remains unclear how fatty type influences the effects of bacterial endotoxins.HTR-8/SVneo trophoblasts were cultured in either 0.5\xa0mmol\xa0L (PA) or linoleic (LA) in the absence or presence of 100\xa0μg\xa0mL of lipopolysaccharide (LPS) or lipoteichoic (LTA). Murine placental explants were cultured in either 2\xa0mmol\xa0L PA or LA, and cell viability, total antioxidant capacity (TAC), lipid peroxidation, H O , heme oxygenase-1 (HO-1), and nuclear erythroid 2-related factor 2 (Nrf-2) and nuclear factor-kappa light-chain enhancer of activated B cells (NF-κB) transcription factor activity assays were assessed. significantly (i) increased cell death, (ii) decreased TAC, and (iii) increased lipid peroxidation; but did not significantly increase HO-1. In contrast, LA maintained cell viability and significantly increased TAC and HO-1. In addition, incubating placental explants with PA significantly increased NF-κB activity. Co-incubating cells with PA and LPS or LTA significantly potentiated H O production and increased lipid peroxidation. Co-incubating cells with PA and LTA synergistically impaired TAC, and LTA decreased TAC more so than LPS. Co-incubation with PA/LA and LPS/LTA decreased HO-1 levels compared to treatment with either fatty alone.Our findings suggest that saturated and unsaturated fats differentially regulate placental viability, antioxidant capacity, and and the actions of gram-positive and gram-negative endotoxins.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: inflammation

is an intracellular signaling molecule involved in disease development.

Emerging evidence shows that (PA), a common fatty in the human diet, serves as a signaling molecule regulating the progression and development of many diseases at the molecular level. In this review, we focus on its regulatory roles in the development of five pathological conditions, namely, metabolic syndrome, cardiovascular diseases, cancer, neurodegenerative diseases, and . We summarize the clinical and epidemiological studies; and also the mechanistic studies which have identified the molecular targets for PA in these pathological conditions. Activation or inactivation of these molecular targets by PA controls disease development. Therefore, identifying the specific targets and signaling pathways that are regulated by PA can give us a better understanding of how these diseases develop for the design of effective targeted therapeutics.

Keyword: inflammation

Docosahexaenoic reversed atherosclerotic changes in human endothelial cells induced by in vitro.

Abnormal activity of atherosclerotic endothelial cells paving luminal surface of blood vessels has been described in many diseases. It has been reported that natural polyunsaturated fatty acids such as docosahexaenoic exert therapeutic effects in atherosclerotic condition. Human umbilical vein endothelial cells were treated with 1mM for 48\xa0hours and exposed to 40μM docosahexaenoic for the next 24 hours. Real-time polymerase chain reaction analysis was used to measure the expression of PTX3, iNOS, and eNOS. The level of nitric oxide was detected by Griess reagent. The transcription level of genes participating in coagulation and blood pressure was studied by polymerase chain reaction array. Docosahexaenoic improved the survival rate by reducing apoptosis rate (P\xa0<\xa0.05). Compared with that of the group given , attenuation of proinflammatory status was indicated by reduced interleukin-6 (P\xa0<\xa0.05) and prostaglandin E levels. All genes PTX3, iNOS, and eNOS were down-regulated after being exposed to docosahexaenoic . Nitric oxide contents were not changed in cells exposed to docosahexaenoic . Polymerase chain reaction array confirmed the reduction of LPA, PDGFβ, ITGA2, SERPINE1, and FGA after exposure to docosahexaenoic for 24\xa0hours (P\xa0<\xa0.05). Docosahexaenoic had potential to blunt atherosclerotic changes in the modulation of genes controlling blood coagulation, pressure, and platelet function.The current experiment showed that docosahexaenoic could reverse atherosclerotic changes in human endothelial cells induced by . The increased levels of interleukin-6 and prostaglandin E in atherosclerotic cells were returned to near-to-normal status. Gene expression analysis showed a reduced activity of genes participating in atherosclerotic endothelial cells treated by docosahexaenoic . The expression of genes related to cell clotting activity was also similar to that of normal cells.Copyright © 2018 John Wiley & Sons, Ltd.

Keyword: inflammation

Priming of Hypothalamic Ghrelin Signaling and Microglia Activation Exacerbate Feeding in Rats\' Offspring Following Maternal Overnutrition.

Maternal overnutrition during pregnancy leads to metabolic alterations, including obesity, hyperphagia, and in the offspring. Nutritional priming of central and its role in ghrelin sensitivity during fed and fasted states have not been analyzed. The current study aims to identify the effect of maternal programming on microglia activation and ghrelin-induced activation of hypothalamic neurons leading to food intake response. We employed a nutritional programming model exposing female Wistar rats to a cafeteria diet (CAF) from pre-pregnancy to weaning. Food intake in male offspring was determined daily after fasting and subcutaneous injection of ghrelin. Hypothalamic ghrelin sensitivity and microglia activation was evaluated using immunodetection for Iba-1 and c-Fos markers, and Western blot for TBK1 signaling. Release of TNF-alpha, IL-6, and IL-1β after stimulation with , oleic, linoleic , or C6 ceramide in primary microglia culture were quantified using ELISA. We found that programmed offspring by CAF diet exhibits overfeeding after fasting and peripheral ghrelin administration, which correlates with an increase in the hypothalamic Iba-1 microglia marker and c-Fos cell activation. Additionally, in contrast to oleic, linoleic, or C6 ceramide stimulation in primary microglia culture, stimulation with for 24 h promotes TNF-alpha, IL-6, and IL-1β release and TBK1 activation. Notably, intracerebroventricular (i.c.v.) or LPS inoculation for five days promotes daily increase in food intake and food consumption after ghrelin administration. Finally, we found that i.c.v. substantially activates hypothalamic Iba-1 microglia marker and c-Fos. Together, our results suggest that maternal nutritional programing primes ghrelin sensitivity and microglia activation, which potentially might mirror hypothalamic administration of the saturated .

Keyword: inflammation

Omega-3-carboxylic acids provide efficacious anti-inflammatory activity in models of crystal-mediated .

This study assesses the efficacy and exposure-response relationship of omega-3-carboxylic acids (OM-3 CA) in models of crystal-based . Human THP-1 macrophages and primary peripheral blood mononuclear cells exposed to multiple inflammatory crystal types were used to determine the anti-inflammatory potential of omega-3 (OM-3) fatty acids in vitro. Anti-inflammatory effects of OM-3 CA in vivo were tested in rat monosodium urate (MSU) crystal air pouch and rat knee intra-articular MSU injection models. Acute treatment with the OM-3 fatty docosahexaenoic suppressed MSU-, cholesterol crystal-, and calcium pyrophosphate crystal-mediated interleukin-1β (IL-1β) production in vitro. In vivo, OM-3 CA dose-dependently reduced crystal-mediated cell migration, exudate volume, and levels of IL-1β and prostaglandin E. Following intra-articular injection of MSU, treatment with OM-3-CA (1\u2009mL/kg) and indomethacin (1\u2009mg/kg) resulted in similar mean reductions in pain (23% and 41%, respectively) and swelling (58% and 50%, respectively), compared with controls. Additionally, in complex formulations of OM-3 fatty acids, high levels of could reduce the in vivo effect on crystal-mediated IL-1β elevation. OM-3 CA has a broadly efficacious anti-inflammatory effect with a strong exposure-response relationship that could be beneficial in prevention and treatment of crystal arthritis, with potential applications in other IL-1β-mediated diseases.

Keyword: inflammation

Caprylic suppresses via TLR4/NF-κB signaling and improves atherosclerosis in ApoE-deficient mice.

As reported previously by our group, medium-chain triglycerides can ameliorate atherosclerosis. Given that TLR4 is closely related to atherosclerosis, we hypothesized herein that caprylic (C8:0) would suppress via TLR4/NF-κB signaling and further promote the amelioration of atherosclerosis in apoE- deficient (apoE) mice.Fifty 6-week male apoE mice were randomly allocated into five diet groups: a high-fat diet (HFD) without or with 2% caprylic (C8:0), capric (C10:0), stearic (C18:0), or linolenic (C18:3). RAW246.7 cells were treated with caprylic (C8:0), docosahexenoic (DHA), (C16:0), and lipopolysaccharide (LPS) with or without TLR4 knock-down (TLR4-KD). The serum lipid profiles, inflammatory biomolecules, and mRNA and protein expression levels were measured. Atherosclerotic lesions that occurred in the aorta and aortic sinuses were evaluated and quantified.Our results indicated that C8:0 reduced body fat, improved the lipid profiles, suppressed inflammatory cytokine production, downregulated aortic TLR4, MyD88, NF-κB, TNF-α, IKKα, and IKKβ mRNA expression, and alleviated atherosclerosis in the apoE mice (\u2009<\u20090.05). In RAW 264.7 cells, C8:0 diminished the inflammatory response and both mRNA and protein expression of TLR4, MyD88, NF-κB, and TNF-α compared to those in the LPS and C16:0 groups (\u2009<\u20090.05). However, in the TLR4-KD RAW 264.7 cells, C8:0 significantly upregulated NF-κB mRNA and protein expression compared to those in the C16:0 and DHA groups.These results suggest that C8:0 functions via TLR4/NF-κB signaling to improve the outcomes of apoE mice through suppressing and ameliorating atherosclerosis. Thus, C8:0 may represent as a promising nutrient against chronic inflammatory diseases.

Keyword: inflammation

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting lipogenesis and .

Non-alcoholic fatty liver disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited lipid accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), fatty synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased lipid accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: inflammation

Exosomes derived from mangiferin‑stimulated perivascular adipose tissue ameliorate endothelial dysfunction.

Perivascular adipose tissue (PVAT) is considered to serve a vital role during the development of endothelial dysfunction. The current study investigated the effect of exosomes derived from mangiferin‑stimulated PVAT on endothelial function, including regeneration, migration, apoptosis and . The number of exosomes secreted by PVAT was increased by stimulation with mangiferin (0.1,\xa01\xa0or\xa010\xa0µM), and uptake of these exosomes by endothelial cells was observed. Exosomes produced by stimulation of PVAT with mangiferin reversed the effects of ‑induced endothelial dysfunction following \xa0(PA) treatment. Furthermore, nuclear factor\xa0(NF)‑κB signaling in endothelial cells was significantly increased when treated with PA‑induced PVAT‑derived exosomes, whereas exosomes from the supernatant of PVAT stimulated with mangiferin reduced p65 and p50 phosphorylation levels in the cells, and inhibited p65 transportation to the nucleus. Taken together, the present study demonstrated that exosomes derived from mangiferin‑stimulated PVAT supernatant inhibited ‑induced endothelial dysfunction via modulation of NF‑κB signaling.

Keyword: inflammation

Tibolone attenuates inflammatory response by and preserves mitochondrial membrane potential in astrocytic cells through estrogen receptor beta.

(PA) induces several metabolic and molecular changes in astrocytes, and, it is involved in pathological conditions related to neurodegenerative diseases. Previously, we demonstrated that tibolone, a synthetic steroid with estrogenic, progestogenic and androgenic actions, protects cells from mitochondrial damage and morphological changes induced by PA. Here, we have evaluated which estrogen receptor is involved in protective actions of tibolone and analyzed whether tibolone reverses gene expression changes induced by PA. Tibolone actions on astrocytic cells were mimicked by agonists of estrogen receptor α (ERα) and β (ERβ), but the blockade of both ERs suggested a predominance of ERβ on mitochondria membrane potential. Expression analysis showed a significant effect of tibolone on genes associated with such as IL6, IL1B and miR155-3p. It is noteworthy that tibolone attenuated the increased expression of TERT, TERC and DNMT3B genes induced by . Our results suggest that tibolone has anti-inflammatory effects and can modulate pathways associated with DNA methylation and telomeric complex. However, future studies are needed to elucidate the role of epigenetic mechanisms and telomere-associated proteins on tibolone actions.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: inflammation

[Atherosclerosis and atheromatosis are consequtive metabolic disordes. Pathology of the biological functions of trophology and endoecology is the basis for ischemic heart disease prevention.]

Atherosclerosis and atheromatosis are different nonphysiological processes with different etiology and pathogenesis. They manifest alterations in different biological functions. According to our original phylogenetic theory of general pathology, atherosclerosis is associated with altered biological function of trophology, eating, biological reaction of exotrophy. Atherosclerosis is induced by eating of nonoptimal for phylogenetically herbivorous Homo sapiens meat diet with high content of saturated fatty (SFA), which leads to in vivo formation of phylogenetically early low-efficient pathway of FA metabolism instead of highly-efficient oleic pathway operating in herbivores. Accumulation of nonligand very low density lipoproteins (VLDL) and low density lipoproteins (LDL) in the bloodstream results from nonphysiological reaction of compensation upon transport of SFA to cells. An increase in blood content of triglycerides (TG) and nonligand VLDLЛ→LDL coincides with the development of hypercholesterolemia: type IV→ type IIb → type V. Atheromatosis compensates changes in lipoproteins by activation of the biological function of endoecology (purity of the extracellular medium) in vivo, thus fulfilling the biological reaction of . This is physiological denaturation of apoВ-100 in nonligand VLDL→LDL by neutrophils via peroxidation, opsonization by the complement components, transcytosis across the endothelial monolayer and removal to the intima of elastic arteries that serves as a collection and utilization pool for phogogens from local intravascular pool of the intercellular medium. Endogenous phlogogens are utilized by phylogenetically early polyfunctional resident macrophages which are small in number and do not proliferate. Blood-borne monocytes-macrophages are also involved in this process, however, they do not express hydrolase of polyenic cholesteryl esters. Atheromatous masses are partially catabolized polyenic FA esterified by the alcohol cholesterol which were not internalized by cells. Atheromatosis is a process of pathological compensation in the realization of the function of endoecology. Prevention of atherosclerosis and atheromatosis should be based on elimination of the effects produced by a nonphysiological meat diet.

Keyword: inflammation

Effects of a new compound containing Palmitoylethanolamide and Baicalein in myocardial ischaemia/reperfusion injury in vivo.

Myocardial ischemia/reperfusion (I/R) injury is the principal cause of death, happens after prolonged obstruction of the coronary arteries. \xa0The first intervention to limit myocardial damage is directed to restoration of perfusion, to avoid inflammatory response and a significant oxidative stress triggered by infarction. Palmitoylethanolamide (PEA), is a well-known fatty amide-signaling molecule that possess an important anti-inflammatory and analgesic effects. PEA does not hold the ability to inhibit free radicals formation. Baicalein, a bioactive component isolated from a Chinese herbal medicine, has multiple pharmacological activities, such as a strong anti-oxidative effects.A combination of PEA and Baicalein could have beneficial effects on oxidative stress produced by inflammatory response.In the present study we explored the effects of composite containing PEA and Baicalein in a model of myocardial I/R injury.Myocardial ischemia/reperfusion injury was induced by occlusion of the left anterior descending coronary artery for 30\u202fmin followed by 2\u202fh of reperfusion. PEA-Baicalein (9:1), was administered (10\u202fmg/kg) 5\u202fmin before the end of ischemia and 1\u202fh after reperfusion.In this study, we clearly demonstrated that PEA-Baicalein treatment decreases myocardial tissue injury, neutrophils infiltration, markers for mast cell activation expression as chymase and tryptase and pro-inflammatory cytokines production (TNF-α, IL-1β). Moreover, PEA-Baicalein treatment reduces stress oxidative and modulates Nf-kB and apoptosis pathways.These results support the idea that the association between PEA and Baicalein should be a potent candidate for the treatment of myocardial I/R injury.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: inflammation

Acetyl-CoA from -induced fatty acids oxidation promotes hepatic malate-aspartate shuttle activity and glycolysis.

Hepatic metabolic syndrome is associated with , as stimulates the reprogramming of nutrient metabolism and hepatic mitochondria-generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and fatty acids, in the condition of is still unclear. Here, we used an acute model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic glycolysis and fatty oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs liver showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain glycolysis. With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived fatty oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by fatty oxidation promotes MAS activity and glycolysis via nonenzymatic acetylation during the inflammatory stress response.

Keyword: inflammation

Sulforaphane ameliorates high-fat diet-induced spermatogenic deficiency in mice†.

Sulforaphane (SFN), a dietary isothiocyanate that is mainly found in cruciferous vegetables, possesses anti-oxidative and anticancer activity and modulates . However, little is known about the role of SFN in obesity-related male reproductive defects. The present study aimed to investigate the effects of SFN on high-fat diet (HFD)-induced male spermatogenic impairment and further clarify the possible underlying mechanisms. In this study, 8-week-old mice were randomly divided into four groups. Mice were fed a normal diet or an HFD with or without SFN supplementation. Sulforaphane was subcutaneously injected at a dose of 0.5 mg/kg 5 days/week for 4 weeks beginning 8 weeks after initiation of the HFD. The results demonstrated that SFN could protect against HFD-induced reproductive dysfunction in male mice. Moreover, SFN also improved reproductive ability, as demonstrated by an increased pregnancy rate and decreased embryo resorption rate in comparison to the corresponding HFD group. We also observed a decrease in apoptosis and an attenuation of endoplasmic reticulum (ER) stress after SFN treatment. In vitro studies of mouse and human sperm samples also revealed that SFN protects against the -induced reduction in sperm viability and motility by inhibiting ER stress in an AMP-activated protein kinase (AMPK)-dependent manner. AMPK-dependent ER stress attenuation by SFN was further confirmed using AMPK knockout mice. Taken together, these data show that SFN protects against HFD-induced male reproductive dysfunction by inhibiting ER stress and apoptosis. These findings may be helpful for identifying new therapeutic methods to treat male infertility.© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

Keyword: inflammation

Mitochondrial Metabolic Reprogramming by CD36 Signaling Drives Macrophage Inflammatory Responses.

A hallmark of chronic inflammatory disorders is persistence of pro-inflammatory macrophages in diseased tissues. In atherosclerosis this is associated with dyslipidemia and oxidative stress, but mechanisms linking these phenomena to macrophage activation remain incompletely understood. To investigate mechanisms linking dyslipidemia, oxidative stress and macrophage activation through modulation of immunemetabolism, and to explore therapeutic potential targeting specific metabolic pathways. Using a combination of biochemical, immunological, and ex vivo cell metabolic studies, we report that CD36 mediates a mitochondrial metabolic switch from oxidative phosphorylation to superoxide production in response to its ligand, oxLDL. Mitochondrial-specific inhibition of superoxide inhibited oxLDL-induced NF-κB activation and inflammatory cytokine generation. RNAseq, flow cytometry, H-labeled uptake, lipidomic analysis, confocal and EM imaging, and functional energetics revealed that oxLDL upregulated effectors of long-chain fatty (FA) uptake and mitochondrial import, while downregulating FA oxidation and inhibiting ATP5A, an electron transport chain (ETC) component. The combined effect is long-chain FA accumulation, alteration of mitochondrial structure and function, repurposing of the ETC to superoxide production, and NF-κB activation. Apoe null mice challenged with high fat diet showed similar metabolic changes in circulating Ly6C monocytes and peritoneal macrophages, along with increased CD36 expression. Moreover, mitochondrial ROS was positively correlated with CD36 expression in aortic lesional macrophages. These findings reveal that oxLDL/CD36 signaling in macrophages links dys-regulated FA metabolism to oxidative stress from the mitochondria, which drives chronic inflammation. Thus, targeting to CD36 and its downstream effectors may serve as potential new strategies against chronic inflammatory diseases such as atherosclerosis.

Keyword: inflammation

Soluble epoxide hydrolase derived lipid mediators are elevated in bronchoalveolar lavage fluid from patients with sarcoidosis: a cross-sectional study.

Sarcoidosis is a systemic inflammatory multi-organ disease almost always affecting the lungs. The etiology remains unknown, but the hallmark of sarcoidosis is formation of non-caseating epithelioid cells granulomas in involved organs. In Scandinavia, >\u200930% of sarcoidosis patients have Löfgren\'s syndrome (LS), an acute disease onset mostly indicating a favorable prognosis. The impact of dysregulation of lipid mediators, which has been investigated in other inflammatory disorders, is still unknown.Using three different liquid chromatography coupled to tandem mass spectrometry targeted platforms (LC-MS/MS), we quantified a broad suite of lipid mediators including eicosanoids, sphingolipids and endocannabinoids in bronchoalveolar lavage (BAL) fluid from pulmonary sarcoidosis patients (n\u2009=\u200941) and healthy controls (n\u2009=\u200916).A total of 47 lipid mediators were consistently detected in BAL fluid of patients and controls. After false discovery rate adjustment, two products of the soluble epoxide hydrolase (sEH) enzyme, 11,12-dihydroxyeicosa-5,8,14-trienoic (11,12-DiHETrE, p\u2009=\u20094.4E-5, q\u2009=\u20091.2E-3, median fold change\u2009=\u20096.0) and its regioisomer 14,15-dihydroxyeicosa-5,8,11-trienoic (14,15-DiHETrE, p\u2009=\u20093.6E-3, q\u2009=\u20093.2E-2, median fold change\u2009=\u20091.8) increased in patients with sarcoidosis. Additional shifts were observed in sphingolipid metabolism, with a significant increase in -derived sphingomyelin (SM16:0, p\u2009=\u20091.3E-3, q\u2009=\u20091.7E-2, median fold change\u2009=\u20091.3). No associations were found between these 3 lipid mediators and LS, whereas levels of SM 16:0 and 11,12-DiHETrE associated with radiological stage (p\u2009<\u20090.05), and levels of 14,15-DiHETrE were associated with the BAL fluid CD4/CD8 ratio.These observed shifts in lipid mediators provide new insights into the pathobiology of sarcoidosis and in particular highlight the sEH pathway to be dysregulated in disease.

Keyword: inflammation

Interactions of fatty acids, nonsteroidal anti-inflammatory drugs, and coxibs with the catalytic and allosteric subunits of cyclooxygenases-1 and -2.

Prostaglandin endoperoxide H synthases-1 and -2, commonly called cyclooxygenases-1 and -2 (COX-1 and -2), catalyze the committed step in prostaglandin biosynthesis-the conversion of arachidonic to prostaglandin endoperoxide H Both COX isoforms are sequence homodimers that function as conformational heterodimers having allosteric (Eallo) and catalytic (Ecat) subunits. At least in the case of COX-2, the enzyme becomes folded into a stable Eallo/Ecat pair. Some COX inhibitors ( nonsteroidal anti-inflammatory drugs and coxibs) and common fatty acids (FAs) modulate Ecat activity by binding Eallo. However, the interactions and outcomes often differ between isoforms. For example, naproxen directly and completely inhibits COX-1 by binding Ecat but indirectly and incompletely inhibits COX-2 by binding Eallo. Additionally, COX-1 is allosterically inhibited up to 50% by common FAs like , whereas COX-2 is allosterically activated 2-fold by . FA binding to Eallo also affects responses to COX inhibitors. Thus, COXs are physiologically and pharmacologically regulated by the FA tone of the milieu in which each operates-COX-1 in the endoplasmic reticulum and COX-2 in the Golgi apparatus. Cross-talk between Eallo and Ecat involves a loop in Eallo immediately downstream of Arg-120. Mutational studies suggest that allosteric modulation requires a direct interaction between the carboxyl group of allosteric effectors and Arg-120 of Eallo; however, structural studies show some allosterically active FAs positioned in COX-2 in a conformation lacking an interaction with Arg-120. Thus, many details about the biological consequences of COX allosterism and how ligand binding to Eallo modulates Ecat remain to be resolved.© 2019 Smith and Malkowski.

Keyword: inflammation

Serum C16:1n7/C16:0 ratio as a diagnostic marker for non-alcoholic steatohepatitis.

Accurate diagnosis of non-alcoholic steatohepatitis (NASH) from non-alcoholic fatty liver disease (NAFLD) is clinically important. Therefore, there is a need for easier ways of diagnosing NASH. In this study, we investigated the serum fatty composition and evaluated the possibility of using the serum fatty composition as a diagnostic marker of NASH.The subjects were 78 NAFLD patients (non-alcoholic fatty liver [NAFL]: 30, NASH: 48) and 24 healthy individuals. Fatty acids extracted from the liver tissue and serum were identified and quantified by gas chromatography. In addition, we evaluated the relationship between serum and liver tissue fatty composition, patient background, and liver histology. The diagnostic performance of NASH was evaluated by calculating the area under the receiver operating characteristic (AUROC).The results of the fatty analysis showed the C16:1n7/C16:0 ratio to have the strongest correlation between serum and liver tissue (r\xa0=\xa00.865, P\xa0<\xa00.0001). The serum C16:1n7/C16:0 ratio in the NASH group was higher compared with that in the NAFL group (P\xa0=\xa00.0007). Evaluation of the association of the serum C16:1n7/C16:0 ratio with liver histology revealed significant correlation with lobular score, ballooning score, and fibrosis score. The AUROC for predicting NASH in all NAFLD patients was 0.7097. The AUROC was nearly equivalent even when the study subjects were restricted to patients with a fibrosis score\xa0≤\xa02 only (AUROC 0.6917).Measuring the serum C16:1n7/C16:0 ratio may be an effective non-invasive method for diagnosing NASH, particularly in its early stages.© 2019 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Keyword: inflammation

Saturated fatty acids, obesity, and the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in asthmatic patients.

Both obesity and high dietary fat intake activate the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome.We aimed to examine NLRP3 inflammasome activity in the airways of obese asthmatic patients after macronutrient overload and in immune cells challenged by inflammasome triggers.Study 1 was a cross-sectional observational study of nonobese (n\xa0=\xa051) and obese (n\xa0=\xa076) asthmatic adults. Study 2 was a randomized, crossover, acute feeding study in 23 asthmatic adults (n\xa0=\xa012 nonobese and n\xa0=\xa011 obese subjects). Subjects consumed 3 isocaloric meals on 3 separate occasions (ie, saturated fatty , n-6 polyunsaturated fatty , and carbohydrate) and were assessed at 0 and 4\xa0hours. For Studies 1 and 2, airway was measured based on sputum differential cell counts, IL-1β protein levels (ELISA), and sputum cell gene expression (Nanostring nCounter). In Study 3 peripheral blood neutrophils and monocytes were isolated by using Ficoll density gradient and magnetic bead separation and incubated with or without , LPS, or TNF-α for 24\xa0hours, and IL-1β release was measured (ELISA).In Study 1 NLRP3 and nucleotide oligomerization domain 1 (NOD1) gene expression was upregulated, and sputum IL-1β protein levels were greater in obese versus nonobese asthmatic patients. In Study 2 the saturated fatty meal led to increases in sputum neutrophil percentages and sputum cell gene expression of Toll-like receptor 4 (TLR4) and NLRP3 at 4\xa0hours in nonobese asthmatic patients. In Study 3 neutrophils and monocytes released IL-1β when challenged with a combination of and LPS or TNF-α.The NLRP3 inflammasome is a potential therapeutic target in asthmatic patients. Behavioral interventions that reduce fatty exposure, such as weight loss and dietary saturated fat restriction, warrant further exploration.Copyright © 2018 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Keyword: inflammation

Low eicosapentaenoic and gamma-linolenic levels in breast adipose tissue are associated with inflammatory breast cancer.

Since it is thought that breast adipose tissue could influence breast cancer clinical presentation, we wanted to characterize specifically the relationship between breast adipose tissue fatty profile and Inflammatory Breast cancer (IBC).Two hundred thirty-four women presenting with breast cancer were managed in our centre between January 2009 and December 2011. Breast adipose tissue specimens were collected during breast surgery. We established the biochemical profile of adipose tissue fatty acids (FA) by gas chromatography and assessed whether there were differences in function of the presence of breast or not.We found that IBC was associated with decreased levels in breast adipose tissue of eicosapentaenoic (EPA), one of the two main polyunsaturated n-3 fatty acids (n-3 PUFA) of marine origin, but also with decreased levels of Gamma Linolenic (GLA). Inversely, an increase in levels was associated with IBC.These differences in lipid content may contribute to the occurrence of breast cancer .Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular in HepG2 Cells.

Fatty acids induced hepatic plays an important role in nonalcoholic fatty liver disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti- in various human organs. However, the anti- property of HS in the fatty liver is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free fatty acids-induced . The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: inflammation

High-fat diet feeding and increase CRC growth in β2AR-dependent manner.

Epidemiology studies indicate that consumption of high-fat diet (HFD) is directly associated with the development of colorectal cancer (CRC). However, the exact component in HFD and the mechanism underlying its effect on CRC growth remained unclear. Our study shows that HFD feeding increases β2AR expression in the xenograft tissues of CRC-bearing mouse model; the elevated β2AR expression is reduced when HFD is replaced by control diet, which strongly suggests an association between HFD feeding and β2AR expression in CRC. HFD feeding increases and stearic levels in CRC; however, only increases β2AR expression, which is dependent upon Sp1. β2AR plays the dominant role in promoting CRC cell proliferation among all the β-AR subtypes. More importantly, knockout of β2AR or knockdown of Sp1 abolishes the increased CRC cell proliferation, suggesting increases CRC cell proliferation in β2AR-dependent manner. HFD or -rich diet (PAD) also fails to increase the tumor growth in xenograft mouse models bearing β2AR-knockout CRC cells. β2AR promotes CRC growth by increasing the phosphorylation of HSL at the residue S552. The phosphorylated and activated HSL (S552) changes the metabolic phenotype of CRC and increases energy production, which promotes CRC growth. Our study has revealed the unique tumorigenic properties of in promoting CRC growth, and have delineated the underlying mechanism of action. We are also the first to report the linkage between HFD feeding and β-adrenergic signaling pathway in relation to CRC growth.

Keyword: inflammation

Human relaxin-2 attenuates hepatic steatosis and fibrosis in mice with non-alcoholic fatty liver disease.

Human relaxin-2 reduces hepatic fibrosis in mice. However, the effects of relaxin-2 on hepatic steatosis and fibrosis in animals with non-alcoholic fatty liver disease (NAFLD) remain to be elucidated. C57BL/6 mice fed a high-fat diet (HFD) or methionine-choline-deficient (MCD) diet were randomly assigned to receive recombinant human relaxin-2 (25 or 75\u2009μg/kg/day) or vehicle for 4 weeks. In HFD-fed mice, relaxin-2 decreased systemic insulin resistance and reduced body weight, epididymal fat mass and serum leptin and insulin concentrations. In livers of HFD-fed mice, relaxin-2 attenuated steatosis and increased phosphorylation of insulin receptor substrate-1, Akt and endothelial nitric oxide synthase (eNOS), and activated genes that regulate fatty oxidation and suppressed acetyl-CoA carboxylase. Relaxin-2 had no direct anti-steatotic effect on primary mouse hepatocytes, but S-nitroso-N-acetylpenicillamine attenuated -induced steatosis and activated genes regulating fatty oxidation in hepatocytes. In mice fed an MCD diet, relaxin-2 attenuated steatosis, and fibrosis. Relaxin-2 increased eNOS and Akt phosphorylation and transcript levels of cytochrome P450-4a10 and decreased acetyl-CoA carboxylase in MCD-fed mouse livers. Moreover, expression levels of Kupffer cell activation, hepatic stellate cell activation and hepatocyte apoptosis were decreased in MCD diet-fed mice receiving relaxin-2. In conclusion, relaxin-2 reduces hepatic steatosis by activating intrahepatic eNOS in HFD-fed mice and further attenuates liver fibrosis in MCD diet-fed mice. Therefore, human relaxin-2 is a potential therapeutic treatment for NAFLD.

Keyword: inflammation

2-Chlorohexadecanoic induces ER stress and mitochondrial dysfunction in brain microvascular endothelial cells.

Peripheral leukocytes induce blood-brain barrier (BBB) dysfunction through the release of cytotoxic mediators. These include hypochlorous (HOCl) that is formed via the myeloperoxidase-HO-chloride system of activated phagocytes. HOCl targets the endogenous pool of ether phospholipids (plasmalogens) generating chlorinated inflammatory mediators like e.g. 2-chlorohexadecanal and its conversion product 2-chlorohexadecanoic (2-ClHA). In the cerebrovasculature these compounds inflict damage to brain microvascular endothelial cells (BMVEC) that form the morphological basis of the BBB. To follow subcellular trafficking of 2-ClHA we synthesized a \'clickable\' alkyne derivative (2-ClHyA) that phenocopied the biological activity of the parent compound. Confocal and superresolution structured illumination microscopy revealed accumulation of 2-ClHyA in the endoplasmic reticulum (ER) and mitochondria of human BMVEC (hCMEC/D3 cell line). 2-ClHA and its alkyne analogue interfered with protein palmitoylation, induced ER-stress markers, reduced the ER ATP content, and activated transcription and secretion of interleukin (IL)-6 as well as IL-8. 2-ClHA disrupted the mitochondrial membrane potential and induced procaspase-3 and PARP cleavage. The protein kinase R-like ER kinase (PERK) inhibitor GSK2606414 suppressed 2-ClHA-mediated activating transcription factor 4 synthesis and IL-6/8 secretion, but showed no effect on endothelial barrier dysfunction and cleavage of procaspase-3. Our data indicate that 2-ClHA induces potent lipotoxic responses in brain endothelial cells and could have implications in -induced BBB dysfunction.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: inflammation

The BACE1 product sAPPβ induces ER stress and and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, , and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and fatty oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, , insulin resistance, PGC-1α down-regulation, and fatty oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, and insulin resistance in skeletal muscle and gluconeogenesis in liver.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Chronic intake of moderate fat-enriched diet induces fatty liver and low-grade without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with . Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of -related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced fatty liver. We also observed the development of low-grade characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: inflammation

Green Tea Polyphenol (-)-Epigallocatechin Gallate (EGCG) Attenuates Neuroinflammation in -Stimulated BV-2 Microglia and High-Fat Diet-Induced Obese Mice.

Obesity is closely associated with neuroinflammation in the hypothalamus, which is characterized by over-activated microglia and excessive production of pro-inflammatory cytokines. The present study was aimed at elucidating the effects of (-)-epigallocatechin gallate (EGCG) on -stimulated BV-2 microglia and high-fat-diet-induced obese mice. The results indicated the suppressive effect of EGCG on lipid accumulation, pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) release, and microglial activation in both cellular and high-fat-diet rodent models. These results were associated with lower phosphorylated levels of the janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) signaling pathway. In conclusion, EGCG can attenuate high-fat-induced hypothalamic via inhibiting the JAK2/STAT3 signaling pathways in microglia.

Keyword: inflammation

Profiling of epidermal lipids in a mouse model of dermatitis: Identification of potential biomarkers.

Lipids are important structural and functional components of the skin. Alterations in the lipid composition of the epidermis are associated with and can affect the barrier function of the skin. SHARPIN-deficient cpdm mice develop a chronic dermatitis with similarities to atopic dermatitis in humans. Here, we used a recently-developed approach named multiple reaction monitoring (MRM)-profiling and single ion monitoring to rapidly identify discriminative lipid ions. Shorter fatty acyl residues and increased relative amounts of sphingosine ceramides were observed in cpdm epidermis compared to wild type mice. These changes were accompanied by downregulation of the Fasn gene which encodes fatty synthase. A profile of diverse lipids was generated by fast screening of over 300 transitions (ion pairs). Tentative attribution of the most significant transitions was confirmed by product ion scan (MS/MS), and the MRM-profiling linear intensity response was validated with a C17-ceramide lipid standard. Relative quantification of sphingosine ceramides CerAS(d18:1/24:0)2OH, CerAS(d18:1/16:0)2OH and CerNS(d18:1/16:0) discriminated between the two groups with 100% accuracy, while the free fatty acids cerotic , 16-hydroxy , and docosahexaenoic (DHA) had 96.4% of accuracy. Validation by liquid chromatography tandem mass spectrometry (LC-MS/MS) of the above-mentioned ceramides was in agreement with MRM-profiling results. Identification and rapid monitoring of these lipids represent a tool to assess therapeutic outcomes in SHARPIN-deficient mice and other mouse models of dermatitis and may have diagnostic utility in atopic dermatitis.

Keyword: inflammation

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic fatty liver disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), cirrhosis, and cancer. The role of different types of fatty acids in driving the early progression of NAFL to NASH is not understood. Lipid overload causing lipotoxicity and has been considered as an essential pathogenic factor. To correlate the lipid profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced lipid overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced and cell death, whereas C16:1 induced significant lipid droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. Lipid profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific fatty acids that affect lipid metabolism and cause lipotoxicity or lipid droplet formation. It indicates that not only saturation or monounsaturation of fatty acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of lipid metabolism and the heterogeneous development of NAFLD.

Keyword: inflammation

Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and .

Danzhi jiangtang capsule (DJC) is mainly used to treat type 2 diabetes mellitus and vascular complication. Our aim was to investigate whether the protective effects of DJC by oral administration on high-fat diet (HFD) and -induced damages of endothelial cells are related to oxidative stress and endoplasmic reticulum (ER) stress. Male Sprague-Dawley rats were randomly divided into standard chow diet (SCD), HFD, HFD plus DJC-low dose (HFD\u2009+\u2009DJCL) and HFD plus DJC-high dose treatment groups (HFD\u2009+\u2009DJCH). The latter three groups were given HFD feeding for three months, then the HFD\u2009+\u2009DJCL and HFD\u2009+\u2009DJCH rats were treated with DJC (500 and 1000\u2009mg/kg/day) via gavage for another two months respectively. Endothelium-dependent relaxations induced by acetylcholine were observed in isolated rat thoracic aortic ring preparation. Malondialdehyde (MDA), total-antioxidant capacity (T-AOC), superoxide dismutase (SOD), interleukin 1β (IL-1β), tumour necrosis factor α (TNFα), free fatty acids (FFA), endothelin-1 (ET-1) and nitric oxide (NO) levels in serum were assayed. The pathological changes, protein expression of endothelial NO synthase (eNOS), phosphorylated eNOS (p-eNOS) and ER stress-related genes in the thoracic aorta were measured. Human umbilical vein endothelial cells (HUVEC) were treated with serum-medicated DJC and then with to detect the reactive oxygen species (ROS) levels and C/EBP homologous protein (CHOP) distribution, expression of p-eNOS and ER stress-related genes. DJC therapy exhibited a potent antioxidant activity and effectively prevented the endothelial dysfunction (ED) manifested by promoting p-eNOS expression and enhancing NO release, decreasing lipid deposition (Oil-red O staining, CPT1b and ACC) and (IL-1β, TNFα, CD68 and p-JNK), alleviating oxidative and ER stress, and decreasing the apoptosis of endothelial cells (TUNEL, BCL-2 and BAX) induced by HFD and respectively. These findings suggest that the administration of DJC had endothelial protective effects against HFD-induced ED, and apoptosis by alleviating lipid deposition, oxidative and ER stress.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: inflammation

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: inflammation

MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue in obesity. Both MIP-1α and free fatty acids are elevated in obesity/T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling pathways were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling pathways. In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in obesity/T2D may contribute to metabolic through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: inflammation

Signal transducer and activator of transcription-3 drives the high-fat diet-associated prostate cancer growth.

Prostate cancer (PCa) is the second leading cause of cancer death in men. PCa progression can be associated with obesity. Signal transducer and activator of transcription-3 (STAT3) plays a crucial role in PCa growth. However, whether STAT3 plays a role in high-fat diet (HFD)-associated PCa growth is unknown. Our data show that HFD feeding increases tumor size, STAT3 phosphorylation, and (PA) level in the xenograft tissues of the PCa-bearing xenograft mouse model. In vitro studies show that PA increases STAT3 expression and phosphorylation (STAT3-Y705) in PCa. Computational modeling suggests strong and stable binding between PA and unphosphorylated STAT3 at R593 and N538. The binding changes STAT3 structure and activity. Functional studies show that both STAT3 mutants (R583A and N538A) and STAT3 dominant negative significantly reduce PA-enhanced STAT3 phosphorylation, PA-increased PCa cell proliferation, migration, and invasion. In the xenograft mouse models, the HFD-increased tumor growth and STAT3 phosphorylation in tumors are reversed by STAT3 inhibition. Our study not only demonstrates the regulatory role of PA/STAT3 axis in HFD-associated PCa growth but also suggests a novel mechanism of how STAT3 is activated by PA. Our data suggest STAT3 as a therapeutic target for the treatment of HFD-associated PCa.

Keyword: inflammation

Sphingosine-1-phosphate analog FTY720 reverses obesity but not age-induced anabolic resistance to muscle contraction.

Sarcopenia, the age-associated loss of skeletal muscle mass and function, is coupled with declines in physical functioning leading to subsequent higher rates of disability, frailty, morbidity, and mortality. Aging and obesity independently contribute to muscle atrophy that is assumed to be a result of the activation of mutual physiological pathways. Understanding mechanisms contributing to the induction of skeletal muscle atrophy with aging and obesity is important for determining targets that may have pivotal roles in muscle loss in these conditions. We find that aging and obesity equally induce an anabolic resistance to acute skeletal muscle contraction as observed with decreases in anabolic signaling activation after contraction. Furthermore, treatment with the sphingosine-1-phosphate analog FTY720 for 4 wk increased lean mass and strength, and the anabolic signaling response to contraction was improved in obese but not older animals. To determine the role of chronic and different fatty acids on anabolic resistance in skeletal muscle cells, we overexpressed IKKβ with and without exposure to saturated fatty (SFA; ), polyunsaturated fatty (eicosapentaenoic ), and monounsaturated fatty (oleic ). We found that IKKβ overexpression increased markers in muscle cells, and this chronic exacerbated anabolic resistance in response to SFA. Pretreatment with FTY720 reversed the inflammatory effects of in the muscle cells. Taken together, these data demonstrate chronic can induce anabolic resistance, SFA aggravates these effects, and FTY720 can reverse this by decreasing ceramide accumulation in skeletal muscle.

Keyword: inflammation

Evaluation of anti-nociceptive and anti-inflammatory activities of the methanol extract of Holigarna caustica (Dennst.) Oken leaves.

Holigarna caustica (Dennst.) is commonly used in traditional medicine to treat a variety of painful conditions such as eye irritation, , arthritis, skin diseases, cuts and wounds.The present study was undertaken to investigate the anti-nociceptive and anti-inflammatory activities of the methanol extract of H. caustica leaves and to elucidate its possible mechanism(s) of action.Fresh leaves of H. caustica were collected, dried, and extracted with methanol (MEHC). MEHC was subjected to activity testing, using chemical-induced (acetic and formalin test) and heat-induced (hot plate and tail immersion test) pain models. To determine the possible mechanism behind the anti-nociceptive activity of MEHC, the opioid antagonist naltrexone was used to evaluate the involvement of opioid receptors in the case of formalin, hot plate and tail immersion tests, while the involvement of the cGMP and ATP-sensitive K channel pathways were assessed using methylene blue and glibenclamide respectively, in the acetic -induced writhing test. In parallel, the carrageenan-induced paw oedema model was used to determine the anti-inflammatory potential of the extract. Exploratory and motor behaviours were evaluated by the open-field test. Various bioactive compounds potentially responsible for the anti-nociceptive and anti-inflammatory activities were ascertained using GC-MS analysis.MEHC showed strong, significant and dose-dependent anti-nociceptive activity in all chemical-induced and heat-induced pain models at all experimental doses. The association of opioid receptors with the observed anti-nociceptive effects was confirmed by using naltrexone. The cGMP and ATP-sensitive K channel pathway was also shown to be involved in the anti-nociceptive activity of MEHC. In addition, MEHC exhibited a dose-dependent inhibition of inflammatory oedema induced by carrageenan. MEHC was not connected with changes in either the locomotor activity or motor responses of mice. In a GC-MS analysis, 40 compounds were identified, among which twelve are documented bioactive compounds with potent analgesic and anti-inflammatory properties.Our current study revealed that MEHC possesses strong central and peripheral anti-nociceptive as well as anti-inflammatory activity. It may also be concluded that both opioid receptors as well as the cGMP and ATP-sensitive K channel pathway are involved in the anti-nociceptive mechanism of MEHC. This study rationalizes the ethnomedicinal use of H. caustica leaves in various painful conditions.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: inflammation

Loss of toll-like receptor 3 aggravates hepatic but ameliorates steatosis in mice.

The importance of toll-like receptor (TLR) 4 in the pathogenesis of steatohepatitis has been well documented; however, little is known about the role of TLR3. In this study, we determined whether the depletion of TLR3 modulated hepatic injury in mice and further aimed to provide mechanistic insights into the TLR3-mediated modulation of diet-induced hepatic and fat accumulation. Hepatic steatosis and inflammatory response were induced by feeding wild-type (WT) or TLR3 knockout mice a high-fat diet for 8 weeks. Primary liver resident cells, including hepatocytes, Kupffer cells, and hepatic stellate cells (HSCs), were treated with . TLR3 knockout mice fed a high-fat diet showed severe hepatic accompanied by nuclear factor-κB and IRF3 activation, which is mainly induced by the activation of Kupffer cells. Decreased TLR4 expression was restored in hepatic mononuclear cells and Kupffer cells in TLR3 knockout mice compared to that in the WT. Moreover, hepatic steatosis was decreased in TLR3 knockout mice. Hepatocytes from TLR3 knockout mice exhibited reduced expression of cannabinoid receptors. HSCs from TLR3 knockout mice showed decreased expression of the enzymes involved in endocannabinoid synthesis. In conclusion, this study suggests that the selective modulation of TLR3 could be a novel therapeutic target for the treatment of hepatic and steatosis.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: inflammation

Induces Müller Cell that is Potentiated by Co-treatment with Glucose.

Chronic hyperglycemia is thought to be the major stimulator of retinal dysfunction in diabetic retinopathy (DR). Thus, many diabetes-related systemic factors have been overlooked as inducers of DR pathology. Cell culture models of retinal cell types are frequently used to mechanistically study DR, but appropriate stimulators of DR-like factors are difficult to identify. Furthermore, elevated glucose, a gold standard for cell culture treatments, yields little to no response from many primary human retinal cells. Thus, the goal of this project was to demonstrate the effectiveness of the free fatty , and compare its use alone and in combination with elevated glucose as a stimulus for human Müller cells, a retinal glial cell type that is activated early in DR pathogenesis and uniquely responsive to fatty acids. Using RNA sequencing, we identified a variety of DR-relevant pathways, including NFκB signaling and , intracellular lipid signaling, angiogenesis, and MAPK signaling, that were stimulated by , while elevated glucose alone did not significantly alter any diabetes-relevant pathways. Co-treatment of high glucose with potentiated the expression of several DR-relevant angiogenic and inflammatory targets, including PTGS2 (COX-2) and CXCL8 (IL-8).

Keyword: inflammation

Crucial Roles of 5-HT and 5-HT2 Receptor in Diabetes-Related Lipid Accumulation and Pro-Inflammatory Cytokine Generation in Hepatocytes.

Previously, we confirmed that liver-synthesized 5-HT rather than non-liver 5-HT, acting on the 5-HT2 receptor (5-HT2R), modulates lipid-induced excessive lipid synthesis (ELS). Here, we further revealed the effects of the hepatocellular 5-HT system in diabetes-related disorders.Studies were conducted in male ICR mice, human HepG2 cells, and primary mouse hepatocytes (PMHs) under gene or chemical inhibition of the 5-HT system, key lipid metabolism, and -related factors. Protein and messenger RNA expression and levels of the factors were determined via western blotting, reverse transcription PCR, and quantitative assay kits, respectively. Hepatic steatosis with and fibrosis, intracellular lipid droplet accumulation (LDA), and reactive oxygen species (ROS) location were determined via hematoxylin and eosin, Masson\'s trichrome, Oil red O, and fluorescent-specific staining, respectively. induced the activation of the 5-HT system: the activation of 5-HT2R, primarily 5-HT2AR, in addition to upregulating monoamine oxidase A (MAO-A) expression and 5-HT synthesis, by activating the G protein/ phospholipase C pathway modulated PKCε activation, resulting in ELS with LDA; the activation of NF-κB, which mediates the generation of pro-inflammatory cytokines, was primarily due to ROS generation in the mitochondria induced by MAO-A-catalyzed 5-HT degradation, and secondarily due to the activation of PKCε. These effects of the 5-HT system were also detected in - or high glucose-treated PMHs and regulated multiple inflammatory signaling pathways. In diabetic mice, co-treatment with antagonists of both 5-HT synthesis and 5-HT2R significantly abolished hepatic steatosis, , and fibrosis as well as hyperglycemia and dyslipidemia.Activation of the hepatocellular 5-HT system plays a crucial role in inducing diabetes-related hepatic dysfunction and is a potential therapeutic target.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: inflammation

Comprehensive genetic study of fatty acids helps explain the role of noncoding inflammatory bowel disease associated SNPs and fatty metabolism in disease pathogenesis.

Fatty acids and their derivatives play an important role in . Diet and genetics influence fatty profiles. Abnormalities of fatty profiles have been observed in inflammatory bowel diseases (IBD), a group of complex diseases defined by chronic gastrointestinal . IBD associated fatty profile abnormalities were observed independently of nutritional status or disease activity, suggesting a common genetic background. However, no study so far has attempted to look for overlap between IBD loci and fatty associated loci or investigate the genetics of fatty profiles in IBD. To this end, we conducted a comprehensive genetic study of fatty profiles in IBD using iCHIP, a custom microarray platform designed for deep sequencing of immune-mediated disease associated loci. This study identifies 10 loci associated with fatty profiles in IBD. The most significant associations were a locus near CBS (p\u202f=\u202f7.62\u202f×\u202f10) and a locus in LRRK2 (p\u202f=\u202f1.4\u202f×\u202f10). Of note, this study replicates the FADS gene cluster locus, previously associated with both fatty profiles and IBD pathogenesis. Furthermore, we identify 18 carbon chain trans-fatty acids (p\u202f=\u202f1.12\u202f×\u202f10), total trans-fatty acids (p\u202f=\u202f4.49\u202f×\u202f10), (p\u202f=\u202f5.85\u202f×\u202f10) and arachidonic (p\u202f=\u202f8.58\u202f×\u202f10) as significantly associated with IBD pathogenesis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammation

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive oxygen species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect and lipid content in the liver.The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and induced by HF diet , leading to improved metabolic disorder and associated liver injury in a mouse model of non-alcoholic fatty liver disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: inflammation

Oxidative stress induced by modulates K2.3 channels in vascular endothelium.

Elevated plasma free fatty acids level has been implicated in the development of insulin resistance, , and endothelial dysfunction in diabetic and nondiabetic individuals. However, the underlying mechanisms still remain to be defined. Herein, we investigated the effect of (PA), the most abundant saturated fatty in the human body, on small-conductance Ca-activated potassium channels (K2.3)-mediated relaxation in rodent resistance arteries and the underlying molecular mechanism. The effect of PA on K2.3 in endothelium was evaluated using real-time PCR, Western blotting, whole-cell patch voltage-clamp, wire and pressure myograph system, and reactive oxygen species (ROS) were measured by using dihydroethidium and 2\', 7\'-dichlorofluorescein diacetate. K2.3-mediated vasodilatation responses to acetylcholine and NS309 (agonist of K2.3 and K3.1) were impaired by incubation of normal mesenteric arteries with 100\u202fμM\u202fPA for 24\u202fh. In cultured human umbilical vein endothelial cells (HUVECs), PA decreased K2.3 current and expression at mRNA and protein levels. Incubation with the NADPH oxidase (Nox) inhibitor dibenziodolium (DPI) partly inhibited the PA-induced ROS production and restored K2.3 expression. Inhibition of either p38-MAPK or NF-κB using specific inhibitors (SB203580, SB202190 or Bay11-7082, pyrrolidinedithiocarbamate) attenuated PA-induced downregulation of K2.3 and inhibition of p38-MAPK also attenuated PA-induced phosphorylation of NF-κB p65. Furthermore, DPI reversed the increment of phospho-p38-MAPK by PA. These results demonstrated that PA downregulated K2.3 expressions via Nox/ROS/p38-MAPK/NF-κB signaling leading to endothelial vasodilatory dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: inflammation

High-fat diet promotes experimental colitis by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental colitis in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental colitis model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe colitis compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes colitis by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes colitis in a 2,4,6-trinitrobenzenesulfonic -induced experimental colitis model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: inflammation

Palmitoleic has Stronger Anti-Inflammatory Potential in Human Endothelial Cells Compared to Oleic and Acids.

Fatty acids (FAs) may affect endothelial cell (EC) function, influencing atherogenesis and inflammatory processes. Palmitoleic (POA) has been described as an anti-inflammatory FA. However, its effects on ECs are underexplored. This study compares the effects of POA with those of (PA) and oleic (OA) on EC inflammatory responses.EAHy926 cells (EC lineage) are exposed to PA, OA, or POA, and stimulated with tumor necrosis factor (TNF)-α. Associated with the FA\'s own incorporation, PA induces a twofold increase in arachidonic , while POA increases the amount of cis-vaccenic . PA, but not OA, enhances the production of IL-6 and IL-8 in response to TNF-α. In contrast, POA decreases production of monocyte chemotactic protein (MCP)-1, IL-6, and IL-8 compared to PA. TNF-α increases surface intercellular adhesion molecule-1 expression previously decreased by POA. TNF-α stimulation increases the expression of NFκB, cyclooxygenase (COX)-2, MCP-1, and IL-6 genes and reduces the expression of peroxisome proliferator-activated receptor (PPAR)-α gene. PA enhances the expression of MCP-1, IL-6, and COX-2 genes, while POA downregulates these genes, decreases expression of NFκB, and upregulates PPAR-α gene expression.POA has anti-inflammatory effects on ECs stimulated with TNF-α and may counter endothelial dysfunction.© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: inflammation

Oleic ameliorates -induced ER stress and markers in naive and cerulein-treated exocrine pancreas cells.

Dietary fat overload (typical to obesity) increases the risk of pancreatic pathologies through mechanisms yet to be defined. We previously showed that saturated dietary fat induces pancreatic acinar lipotoxicity and cellular stress. The endoplasmic reticulum (ER) of exocrine pancreas cells is highly developed and thus predisposed to stress. We studied the combination of saturated and unsaturated FAs in metabolic and pancreatitis like cerulein (CER)-induced stress states on cellular ER stress.Exocrine pancreas AR42J and rat primary exocrine acinar cells underwent acute (24 h) challenge with different FAs (saturated, monounsaturated) at different concentrations (250 and 500 µM) and in combination with acute CER-induced stress, and were analyzed for fat accumulation, ER stress unfolded protein response (UPR) and immune and enzyme markers. Acute exposure of AR42J and pancreatic acinar cells to different FAs and their combinations increased triglyceride accumulation. significantly dose-dependently enhanced the UPR, immune factors and pancreatic lipase (PL) levels, as demonstrated by XBP1 splicing and elevation in UPR transcripts and protein levels ( and ). Exposure to high levels in a CER-induced stress state synergistically increased ER stress and marker levels. Exposure to oleic did not induce ER stress and PL levels and significantly decreased immune factors in an acute CER-induced stress state. Combination of oleic and acids significantly reduced the -induced ER stress, but did not affect the immune factor response. We show that combination of monounsaturated and saturated FAs protects from exocrine pancreatic cellular ER stress in both metabolic and CER-induced stress.© 2019 The Author(s).

Keyword: inflammation

Impact of miR-140 Deficiency on Non-Alcoholic Fatty Liver Disease.

We have previously shown that loss of miR-140 has a pro-fibrotic effect in the mammary gland. This study aims to investigate whether miR-140 loss and obesity act synergistically to promote non-alcoholic fatty liver disease (NAFLD), and to identify the underlying mechanisms.Liver tissues were isolated from lean-fat-diet and high-fat-diet fed wild-type and miR-140 knockout mice. Using molecular staining and immunohistochemistry techniques, increased development of NAFLD and fibrotic indicators in miR-140 knockout mice were identified. Utilizing an in vitro model system, miR-140 was demonstrated to target TLR-4, and miR-140 overexpression was shown to be sufficient to inhibit signaling through the TLR-4/NFκB pathway.These findings demonstrate that loss of miR-140 results in increased expression of TLR-4, sensitizing cells to signaling and in increased inflammatory activity through the TLR4/NFκB pathway. This signaling axis promotes NAFLD development in a high-fat diet context and indicates the potential utility of miR-140 rescue as a therapeutic strategy in NAFLD.© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: inflammation

Time-Dependent alteration to the structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal microbiota composition, increased intestinal permeability, and onset of type 2 diabetes mellitus (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the microbiota, after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: inflammation

Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4-MD2 complex.

Reactive oxygen species (ROS) contribute to the development of non-alcoholic fatty liver disease. ROS generation by infiltrating macrophages involves multiple mechanisms, including Toll-like receptor 4 (TLR4)-mediated NADPH oxidase (NOX) activation. Here, we show that palmitate-stimulated CD11bF4/80 hepatic infiltrating macrophages, but not CD11bF4/80 Kupffer cells, generate ROS via dynamin-mediated endocytosis of TLR4 and NOX2, independently from MyD88 and TRIF. We demonstrate that differently from LPS-mediated dimerization of the TLR4-MD2 complex, palmitate binds a monomeric TLR4-MD2 complex that triggers endocytosis, ROS generation and increases pro-interleukin-1β expression in macrophages. Palmitate-induced ROS generation in human CD68CD14 macrophages is strongly suppressed by inhibition of dynamin. Furthermore, Nox2-deficient mice are protected against high-fat diet-induced hepatic steatosis and insulin resistance. Therefore, endocytosis of TLR4 and NOX2 into macrophages might be a novel therapeutic target for non-alcoholic fatty liver disease.

Keyword: inflammation

Hepatocyte nuclear receptor SHP suppresses and fibrosis in a mouse model of nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is a burgeoning health problem worldwide, ranging from nonalcoholic fatty liver (NAFL, steatosis without hepatocellular injury) to the more aggressive nonalcoholic steatohepatitis (NASH, steatosis with ballooning, , or fibrosis). Although many studies have greatly contributed to the elucidation of NAFLD pathogenesis, the disease progression from NAFL to NASH remains incompletely understood. Nuclear receptor small heterodimer partner (Nr0b2, ) is a transcriptional regulator critical for the regulation of bile , glucose, and lipid metabolism. Here, we show that SHP levels are decreased in the livers of patients with NASH and in diet-induced mouse NASH. Exposing primary mouse hepatocytes to and lipopolysaccharide , we demonstrated that the suppression of expression in hepatocytes is due to c-Jun N-terminal kinase (JNK) activation, which stimulates c-Jun-mediated transcriptional repression of Interestingly, induction of hepatocyte-specific SHP in steatotic mouse liver ameliorated NASH progression by attenuating liver and fibrosis, but not steatosis. Moreover, a key mechanism linking the anti-inflammatory role of hepatocyte-specific SHP expression to involved SHP-induced suppression of NF-κB p65-mediated induction of chemokine (C-C motif) ligand 2 (CCL2), which activates macrophage proinflammatory polarization and migration. In summary, our results indicate that a JNK/SHP/NF-κB/CCL2 regulatory network controls communications between hepatocytes and macrophages and contributes to the disease progression from NAFL to NASH. Our findings may benefit the development of new management or prevention strategies for NASH.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: inflammation

Epigallocatechin gallate improves insulin resistance in HepG2 cells through alleviating and lipotoxicity.

High levels of circulating free fatty acids (FFAs), and oxidative stress are important causes for insulin resistance (IR) and type 2 diabetes mellitus. The aim of this study was to investigate the mechanisms of EGCG in alleviating IR in HepG2 cells.HepG2 cells were treated with 25\u202fmM glucose, 0.25\u202fmM (PA), or 50\u202fμM EGCG for 24\u202fh.EGCG increased glucose uptake and decreased glucose content. EGCG markedly decreased the levels of inflammatory and oxidative stress factors including nuclear factor κB (NF-κB), tumor necrosis factor-α, interleukin-6, reactive oxygen species, malondialdehyde and p53 protein, and markedly increased superoxide dismutases (SOD), glutathione peroxidase and SOD2 protein. EGCG significantly downregulated the levels of FFAs, triacylglycerol and cholesterol in HepG2 cells. The glucose transporter 2 (GLUT2) protein and its downstream proteins peroxisome proliferator-activated receptor γ coactivator (PGC)-1β were significantly increased, and sterol regulatory element-binding-1c (SREBP-1c) protein, and fatty synthase (FAS) were significantly decreased by EGCG in HepG2. Moreover, the foregoing effects were reversed by siRNA-mediated knockdown of GLUT2.Our data demonstrated that EGCG improved IR, possibly through ameliorating glucose (25\u202fmM) and PA (0.25\u202fmM)-induced , oxidative stress, and FFAs via the GLUT2/PGC-1β/SREBP-1c/FAS pathway in HepG2 cells.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: inflammation

Palmitate induces glycosylation of cyclooxygenase-2 in primary human vascular smooth muscle cells.

Vascular basal cyclooxygenase-2 (COX-2) expression and activity can be induced by endotoxin, hypoxia, or ischemia. During vascular pathologies such as atherosclerosis, increases in COX-2 activity result in prostanoid production, a contributor to the development and progression of vascular leading to unstable atherosclerotic plaques and increased risk for thrombotic events. Recent studies demonstrate that select free fatty acids, such as palmitate, can act as proinflammatory mediators. However, the effect of palmitate on COX-2 expression and activity, and its impact on the development and progression of vascular , are not well elucidated. We investigated the effect of palmitate on COX-2 expression and function in human vascular smooth muscle cells. Cells were treated with palmitate, COX-2 protein levels were assessed using Western analysis, and activity was assessed via ELISA. We observed that palmitate dose-dependently increased COX-2 levels and specifically enhanced band intensity of the COX-2 74 kDa band (slowest migrating band). This response was attenuated by N-linked glycosylation inhibition, suggesting that palmitate impacts expression of the fully activated glycoform of COX-2. Palmitate-induced increases in COX-2 levels correlated with an increase in prostaglandin E production that was also attenuated by a glycosylation inhibitor. Additionally, palmitate altered cell morphology and increased cell density which were reversed by selective COX-2 inhibition. Thus, we conclude that palmitate acts on COX-2 by two separate mechanisms of action in human vascular smooth muscle. It elicits dose-dependent increases in COX-2 protein expression and modulates regulation of COX-2 activity via modification of posttranslational glycosylation.

Keyword: inflammation

The effects of pro- and anti-atherosclerotic factors on intracellular nucleotide concentration in murine endothelial cells.

Endothelial cell activation and dysfunction could lead to endothelial injury that is an important factor in the development of vascular diseases. Vascular injury is strongly associated with disturbed endothelial cell energetics and pyridine nucleotide pool. This study aimed to evaluate the effects of inflammatory stimuli (IL-6, LPS), uric , hyperglycemia, fatty acids, flavonoids, statins and nonsteroidal anti-inflammatory drugs on cellular concentration of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and nicotinamide adenine dinucleotide (NAD) in cultured endothelial cells. Murine-immortalized heart endothelial cells (H5V cells) were treated with different concentrations of pro- and anti-atherosclerotic factors and intracellular concentration of nucleotides were measured using high performance liquid chromatography. Intracellular ATP concentration in H5V cells was not changed by inflammatory stimuli (IL-6 and LPS), uric , glucose, atorvastatin, acetylsalicylic , monounsaturated and polyunsaturated fatty acids. Only high concentration of (1\u2009mM) and kaempferol (>0.1\u2009mM) decreased intracellular ATP concentration. The concentration of intracellular ADP has not been altered by any of tested compounds. In turn, intracellular NAD pool was modified only by polyunsaturated fatty acids and atorvastatin. Linoleic , docosahexaenoic and atorvastatin increased cellular NAD concentration. Tested compounds have a small influence on murine endothelial cell energetics, but polyunsaturated fatty acids and atorvastatin increased intracellular NAD concentration that could be an important protective mechanism against endothelial cell injury.

Keyword: inflammation

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid metabolism and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

Keyword: inflammation

Enhancement of lipid content and inflammatory cytokine secretion in SZ95 sebocytes by suggests a potential link between free fatty acids and acne aggravation.

A relationship between acne and free fatty acids (FFAs) has been suggested recently. However, the effects of FFAs on sebaceous glands are still largely unknown. At the same time, the role of FFAs during chronic is well established. Considering that FFAs are also a major component of sebum, it is likely that changes in FFA affect both the synthesis of sebum and the inflammatory response in sebaceous glands. In this study, we examined a hypothesis that FFAs increase the production of sebum and induce in the sebaceous glands. We found that treatment of SZ95 sebocytes with exogenously applied (PA), a major saturated FFA, induced a significant increase in intracellular lipid levels. Moreover, PA treatment also increased the expression and secretion of the proinflammatory cytokines in SZ95 sebocytes. We also found that Toll-like receptors were required for the inflammatory response triggered by PA. The results of our study strengthen the notion about the link between acne and FFAs and suggest the mechanism underlying this relationship. Our results serve as a foundation for future work that will explore the association between FFA and acne and pave way to the development of novel treatment options for acne.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: inflammation

Saturated Fatty Acids Undergo Intracellular Crystallization and Activate the NLRP3 Inflammasome in Macrophages.

provoked by the imbalance of fatty composition, such as excess saturated fatty acids (SFAs), is implicated in the development of metabolic diseases. Recent investigations suggest the possible role of the NLRP3 (nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3) inflammasome, which regulates IL-1β (interleukin 1β) release and leads to , in this process. Therefore, we investigated the underlying mechanism by which SFAs trigger NLRP3 inflammasome activation.The treatment with SFAs, such as and stearic , promoted IL-1β release in murine primary macrophages while treatment with oleic inhibited SFA-induced IL-1β release in a dose-dependent manner. Analyses using polarized light microscopy revealed that intracellular crystallization was provoked in SFA-treated macrophages. As well as IL-1β release, the intracellular crystallization and lysosomal dysfunction were inhibited in the presence of oleic . These results suggest that SFAs activate NLRP3 inflammasome through intracellular crystallization. Indeed, SFA-derived crystals activated NLRP3 inflammasome and subsequent IL-1β release via lysosomal dysfunction. Excess SFAs also induced crystallization and IL-1β release in vivo. Furthermore, SFA-derived crystals provoked acute , which was impaired in IL-1β-deficient mice.These findings demonstrate that excess SFAs cause intracellular crystallization and subsequent lysosomal dysfunction, leading to the activation of the NLRP3 inflammasome, and provide novel insights into the pathogenesis of metabolic diseases.© 2018 American Heart Association, Inc.

Keyword: inflammation

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only energy metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating obesity-induced .

Keyword: inflammation

Dietary Fatty Acids Amplify Inflammatory Responses to Infection through p38 MAPK Signaling.

Obesity is an important risk factor for severe asthma exacerbations, which are mainly caused by respiratory infections. Dietary fatty acids, which are increased systemically in obese patients and are further increased after high-fat meals, affect the innate immune system and may contribute to dysfunctional immune responses to respiratory infection. In this study we investigated the effects of dietary fatty acids on immune responses to respiratory infection in pulmonary fibroblasts and a bronchial epithelial cell line (BEAS-2B). Cells were challenged with BSA-conjugated fatty acids (ω-6 polyunsaturated fatty acids [PUFAs], ω-3 PUFAs, or saturated fatty acids [SFAs]) +/- the viral mimic polyinosinic:polycytidylic (poly[I:C]) or bacterial compound lipoteichoic (LTA), and release of proinflammatory cytokines was measured. In both cell types, challenge with arachidonic (AA) (ω-6 PUFA) and poly(I:C) or LTA led to substantially greater IL-6 and CXCL8 release than either challenge alone, demonstrating synergy. In epithelial cells, (SFA) combined with poly(I:C) also led to greater IL-6 release. The underlying signaling pathways of AA and poly(I:C)- or LTA-induced cytokine release were examined using specific signaling inhibitors and IB. Cytokine production in pulmonary fibroblasts was prostaglandin dependent, and synergistic upregulation occurred via p38 mitogen-activated protein kinase signaling, whereas cytokine production in bronchial epithelial cell lines was mainly mediated through JNK and p38 mitogen-activated protein kinase signaling. We confirmed these findings using rhinovirus infection, demonstrating that AA enhances rhinovirus-induced cytokine release. This study suggests that during respiratory infection, increased levels of dietary ω-6 PUFAs and SFAs may lead to more severe airway and may contribute to and/or increase the severity of asthma exacerbations.

Keyword: inflammation

Docosahexaenoic and Eicosapentaenoic Acids Prevent Altered-Muc2 Secretion Induced by by Alleviating Endoplasmic Reticulum Stress in LS174T Goblet Cells.

Diets high in saturated fatty acids (FA) represent a risk factor for the development of obesity and associated metabolic disorders, partly through their impact on the epithelial cell barrier integrity. We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus barrier. To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or -9, -6, or -3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic nor -9 and -6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 diabetes and obesity or . These in vitro data remain to be explored in vivo in a context of dietary obesity.

Keyword: inflammatory bowel disease

Novel formulation of solid lipid microparticles of curcumin for anti-angiogenic and anti-inflammatory activity for optimization of therapy of inflammatory bowel disease.

This project was undertaken with a view to optimize the treatment of inflammatory bowel disease through a novel drug delivery approach for localized treatment in the colon. Curcumin has poor aqueous solubility, poor stability in the gastrointestinal tract and poor bioavailability. The purpose of the study was to prepare and evaluate the anti-inflammatory activity of solid lipid microparticles (SLMs) of curcumin for the treatment of inflammatory bowel disease in a -induced rat model by a colon-specific delivery approach.We have developed a novel formulation approach for treating experimental in the rat model. SLMs of curcumin were prepared with various lipids, such as , stearic and soya lecithin, with an optimized percentage of poloxamer 188. The SLMs of curcumin were characterized for particle size, drug content, drug entrapment, in-vitro release, surface morphology and infrared, differential scanning calorimetry and X-ray studies. The colonic delivery system of SLM formulations of curcumin were further investigated for their anti-angiogenic and anti-inflammatory activity using chick embryo and rat models.Particle size, drug content, drug entrapment and in-vitro release studies showed that formulation F4 containing one part stearic and 0.5% surfactant had the smallest diameter of 108 microm, 79.24% entrapment and exhibited excellent in-vitro release characteristics when compared with other formulations and pure curcumin. SLMs of curcumin (F4) proved to be a potent angio-inhibitory compound, as demonstrated by inhibition of angiogenesis in the chorioallantoic membrane assay. Rats treated with curcumin and its SLM complex showed a faster weight gain compared with dextran sulfate solution (DSS) control rats. The increase in whole colon length appeared to be significantly greater in SLM-treated rats when compared with pure curcumin and DSS control rats. An additional finding in the DSS-treated rats was chronic cell infiltration with predominance of eosinophils. Decreased mast cell numbers in the mucosa of the colon of SLMs of curcumin and pure curcumin-treated rats was observed.The degree of caused by administration of DSS was significantly attenuated by colonic delivery of SLMs of curcumin. Being a nontoxic natural dietary product, curcumin could be useful in the therapeutic strategy for inflammatory bowel disease patients.

Keyword: inflammatory bowel disease

HADHA, the alpha subunit of the mitochondrial trifunctional protein, is involved in long-chain fatty -induced autophagy in intestinal epithelial cells.

Genome-wide association studies have identified autophagy-related susceptibility genes for inflammatory bowel disease (IBD); however, whether autophagy regulators can be utilized as therapeutic targets remains unclear. To identify novel microtubule-associated protein 1 light chain 3 (LC3)-interacting proteins in intestinal epithelial cells (IECs), we isolated primary IECs from green fluorescent protein (GFP)-LC3 mice. We performed immunoprecipitation with a GFP antibody and then analyzed co-immunoprecipitates by mass spectrometry. HADHA was identified as an LC3-interacting protein from primary IECs. The HADHA gene encodes the alpha subunit of the mitochondrial trifunctional protein. Given that HADHA catalyzes the last three steps of mitochondrial beta-oxidation of long-chain fatty acids, we investigated whether long-chain fatty acids induce autophagy in IECs. We found that induced autophagy in DLD-1, HT29, and HCT116\xa0cells. HADHA was expressed in not only the mitochondria but also the cytosol. LC3 puncta co-localized with HADHA, which were enhanced by stimulation. However, LC3 puncta did not co-localize with Tom20, suggesting that HADHA was induced to associate with LC3 puncta at sites other than the mitochondria. Thus, HADHA may have extra-mitochondrial functions. Furthermore, we found that induced cell death in IECs, which was accelerated by bafilomycin A and chloroquine. These findings suggested that -induced autophagy supports the survival of IECs. Taken together, these results suggested that HADHA is involved in long-chain fatty -induced autophagy in IECs, thus providing new insights into the pathology of IBD and revealing novel therapeutic targets of IBD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: inflammatory bowel disease

Palmitoylethanolamide normalizes intestinal motility in a model of post-inflammatory accelerated transit: involvement of CB₁ receptors and TRPV1 channels.

Palmitoylethanolamide (PEA), a naturally occurring acylethanolamide chemically related to the endocannabinoid anandamide, interacts with targets that have been identified in peripheral nerves controlling gastrointestinal motility, such as cannabinoid CB1 and CB2 receptors, TRPV1 channels and PPARα. Here, we investigated the effect of PEA in a mouse model of functional accelerated transit which persists after the resolution of colonic inflammation (post-inflammatory irritable bowel syndrome).Intestinal inflammation was induced by intracolonic administration of oil of mustard (OM). Mice were tested for motility and biochemical and molecular biology changes 4 weeks later. PEA, oleoylethanolamide and endocannabinoid levels were measured by liquid chromatography-mass spectrometry and receptor and enzyme mRNA expression by qRT-PCR.OM induced transient and a functional post-inflammatory increase in upper gastrointestinal transit, associated with increased intestinal anandamide (but not 2-arachidonoylglycerol, PEA or oleoylethanolamide) levels and down-regulation of mRNA for TRPV1 channels. Exogenous PEA inhibited the OM-induced increase in transit and tended to increase anandamide levels. had a weaker effect on transit. Inhibition of transit by PEA was blocked by rimonabant (CB1 receptor antagonist), further increased by 5'-iodoresiniferatoxin (TRPV1 antagonist) and not significantly modified by the PPARα antagonist GW6471.Intestinal endocannabinoids and TRPV1 channel were dysregulated in a functional model of accelerated transit exhibiting aspects of post-inflammatory irritable bowel syndrome. PEA counteracted the accelerated transit, the effect being mediated by CB1 receptors (possibly via increased anandamide levels) and modulated by TRPV1 channels.© 2014 The British Pharmacological Society.

Keyword: inflammatory bowel disease

Lactobacillus johnsonii glycolipids, their structure and immunoreactivity with sera from inflammatory bowel disease patients.

Structural studies of the major glycolipids produced by two Lactobacillus johnsonii (LJ) strains, LJ 151 isolated from intestinal tract of healthy mice and LJ 142 isolated from mice with experimentally induced inflammatory bowel disease (IBD), were performed. Two major glycolipids, GL1 and GL2, were present in lipid extracts from L.\xa0johnsonii 142 and 151 strains. Glycolipid GL1 has been identified as β-D-Glcp-(1→6)-α-D-Galp-(1→2)-α-D-Glcp-diglyceride and GL2 as α-D-Galp-(1→2)-α-D-Glcp-diglyceride. The main fatty residues identified by gas-liquid chromatography-mass spectrometry were , stearic and lactobacillic acids. Besides structural elucidation of the major glycolipids, the aim of this study was to determine the immunochemical properties of these glycolipids and to compare their immunoreactivity to that of polysaccharides obtained from the same strains. Sera from rabbits immunized with bacterial cells possessed much higher serological reactivity with polysaccharides than with glycolipids. Inversely, reactivity of the glycolipids with human sera from patients with IBD was much higher than that determined for the polysaccharides, while reactivity of glycolipids with human sera from healthy individuals was much lower than one measured for the polysaccharides. Results indicate that glycoconjugates from Lactobacillus cell wall act as antigens and may represent new IBD diagnostic biomarkers.© 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Keyword: inflammatory bowel disease

Palmatine attenuated dextran sulfate sodium (DSS)-induced via promoting mitophagy-mediated NLRP3 inflammasome inactivation.

Activation of NLRP3 inflammasomes is crucial in the pathological process of Ulcerative (UC), which could be negatively regulated by PINK1/Parkin-driven mitophagy. Palmatine is a herb derived isoquinoline alkaloid with potent anti-inflammatory and anti-bacteria activities. In present study, we evaluated the effect of palmatine on dextran sulfate sodium (DSS)-induced mice and examined whether its effect is exerted by promoting mitophagy-mediated NLRP3 inflammasome inactivation. The result showed that palmatine (40, 100\u2009mg/kg) significantly prevented bodyweight loss and colonic shortening in DSS mice, and reduced the disease activity index and histopathologic score. The levels of MPO, IL-1β, TNF-α and the number of F4/80+ cells in colon of DSS mice were remarkably decreased by palmatine. Moreover, palmatine suppressed NLRP3 inflammasomes activation, but enhanced the expression of the mitophagy-related proteins involving LC3, PINK1 and Parkin in colonic tissue of DSS mice. These effects was consistent with the in vitro data revealing that palmatine inhibited the activation of NLRP3 inflammasomes, while promoted the expression and mitochondrial recruitment of PINK1 and Parkin in THP-1 cell differentiated macrophages. Furthermore, the effect of palmatine on THP-1 cells was neutralized by a mitophagy inhibitor Cyclosporin A (CsA) and PINK1-siRNA. In parallel, CsA significantly attenuated the therapeutic effect of palmatine in DSS mice, illustrating that the anti- effect of palmatine is closely related to mitophagy. Taken together, the current results demonstrated that palmatine protected mice against DSS-induced by facilitating PINK1/Parkin-driven mitophagy and thus inactivating NLRP3 inflammasomes in macrophage.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammatory bowel disease

Fatty Acids as Useful Serological Markers for Crohn's Disease.

We have previously reported that patients with Crohn's disease (CD) have a very specific erythrocyte membrane phospholipid fatty profile. The findings of this study suggest that the activities of enzymes involved in the metabolism of linoleic (LA), that is, delta-6 desaturase, are higher in CD patients than in healthy individuals.We evaluated the utilities of various fatty compositions of the plasma (p-) as new serological markers for CD compared to those of erythrocyte membranes (e-).Fifty CD patients and 50 healthy individuals were enrolled. In both plasma and erythrocyte membranes, the weight percentages of (PA) were significantly higher, while those of LA were significantly lower in CD patients than in controls. Fatty acids with high sensitivity and specificity were p-PA (0.86 and 0.74) and e-PA (0.80 and 0.74). With PA and LA as a CD fatty index (CDFAi), that is, CDFAi = (PA/LA), the sensitivity and specificity of plasma CDFAi (p-CDFAi) and e-CDFAi were 0.80 and 0.80; and 0.82 and 0.88 respectively.In CD patients, various fatty acids were specifically altered in both plasma and erythrocytes, and p-PA and p-CDFAi are potentially useful as new serological markers for CD.© 2017 S. Karger AG, Basel.

Keyword: inflammatory bowel disease

Increased arachidonic levels in phospholipids of human colonic mucosa in .

1. Colonic mucosa from 19 patients with ulcerative colitis, eight with Crohn's and 14 controls were analysed for arachidonic (C20:4), linoleic (C18:2), oleic (C18:1), stearic (C18:0) and (C16:0). 2. Gas-liquid chromatography of lipid extracts showed that arachidonic was significantly higher in ulcerative colitis (19 +/- 4) and Crohn's (20 +/- 3) than in controls (13 +/- 5 micrograms/mg of protein) (means +/- SD). Neither the degree of inflammation nor treatment with sulphasalazine or prednisolone appeared to influence the fatty concentrations. 3. Seventy-five to ninety-five per cent of the arachidonic was found in the phospholipid fraction after separation by thin-layer chromatography. There were no significant changes in the concentrations of the other fatty acids measured, although oleic was lower in . The ratios of oleic to stearic and to were lower in . 4. The alteration in the fatty profile may partly explain the increased synthesis of eicosanoids in colonic mucosa in .

Keyword: inflammatory bowel disease

Adelmidrol, a Palmitoylethanolamide Analogue, as a New Pharmacological Treatment for the Management of Inflammatory Bowel Disease.

Leukocyte infiltration, improved levels of intercellular adhesion molecule 1 (ICAM-1), and oxidative stress in the colon are the principal factors in inflammatory bowel disease. The goal of the current study was to explore the effects of adelmidrol, an analog of the anti-inflammatory fatty amide signaling molecule palmitoylethanolamide, in mice subjected to experimental . Additionally, to clarify whether the protective action of adelmidrol is dependent on the activation of peroxisome proliferator-activated receptors (PPARs), we investigated the effects of a PPARγ antagonist, GW9662, on adelmidrol action. Adelmidrol (10 mg/kg daily, o.s.) was tested in a murine experimental model of induced by intracolonic administration of dinitrobenzene sulfonic . Nuclear factor-κB translocation, cyclooxygenase-2, and phosphoextracellular signal-regulated kinase, as well as tumor necrosis factor-α and interleukin-1β, were significantly increased in colon tissues after dinitrobenzene sulfonic administration. Immunohistochemical staining for ICAM-1, P-selectin, nitrotyrosine, and poly(ADP)ribose showed a positive staining in the inflamed colon. Treatment with adelmidrol decreased diarrhea, body weight loss, and myeloperoxidase activity. Adelmidrol treatment, moreover, reduced nuclear factor-κB translocation, cyclooxygenase-2, and phosphoextracellular signal-regulated kinase expression; proinflammatory cytokine release; and the incidence of nitrotyrosine and poly(ADP)ribose in the colon. It also decreased the upregulation of ICAM-1 and P-selectin. Adelmidrol treatment produced a reduction of Bax and an intensification of Bcl-2 expression. This study clearly demonstrates that adelmidrol exerts important anti-inflammatory effects that are partly dependent on PPARγ, suggesting that this molecule may represent a new pharmacologic approach for inflammatory bowel disease treatment.Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: inflammatory bowel disease

Role of acylethanolamides in the gastrointestinal tract with special reference to food intake and energy balance.

Acylethanolamides (AEs) are a group of lipids occurring in both plants and animals. The best-studied AEs are the endocannabinoid anandamide (AEA), the anti-inflammatory compound palmitoylethanolamide (PEA), and the potent anorexigenic molecule oleoylethanolamide (OEA). AEs are biosynthesized in the gastrointestinal tract, and their levels may change in response to noxious stimuli, food deprivation or diet-induced obesity. The biological actions of AEs within the gut are not limited to the modulation of food intake and energy balance. For example, AEs exert potential beneficial effects in the regulation of intestinal motility, secretion, inflammation and cellular proliferation. Molecular targets of AEs, which have been identified in the gastrointestinal tract, include cannabinoid CB(1) and CB(2) receptors (activated by AEA), transient receptor potential vanilloid type 1 (TRPV1, activated by AEA and OEA), the nuclear receptor peroxisome proliferators-activated receptor-alpha (PPAR-alpha, activated by OEA and, to a less extent, by PEA), and the orphan G-coupled receptors GPR119 (activated by OEA) and GPR55 (activated by PEA and, with lower potency, by AEA and OEA). Modulation of AE levels in the gut may provide new pharmacological strategies not only for the treatment of feeding disorders but also for the prevention or cure of widespread intestinal diseases such as inflammatory bowel disease and colon cancer.

Keyword: inflammatory bowel disease

Modulation in delta 9, delta 6, and delta 5 fatty desaturase activity in the human intestinal CaCo-2 cell line.

We report the influence of media lipids, growth in lipid-poor medium, and cell differentiation on delta 9, delta 6, and delta 5 desaturase activity in the human CaCo-2 enterocyte cell line. We also describe the level of incorporation of (16:0), linoleic (18:2n-6), and eicosapentaenoic (EPA) acids (20:5n-3) and their higher homologues into cytosolic and membrane lipids during long-term (10 days) medium supplementation in fully differentiated 16- to 18-day-old cultures. CaCo-2 monolayers reached confluency by day 6 with subsequent development of microvilli and maximal expression of microvillus membrane sucrose, alkaline phosphatase, and gamma-glutamyltransaminase occurring between days 16 and 23 after plating. There was evidence of the presence and modulation of delta 9, delta 6, and delta 5 desaturase activity (delta 9 > delta 6 > delta 5). delta 6 Desaturase activity decreased approximately 2-fold between days 6 and 24 of culture and when the fetal bovine serum concentration was increased from 0.5% to 25%; in contrast, when cells were starved for 72 h, activity increased 5.4-fold. When the media was supplemented with either linoleic and/or EPA, both delta 6 and delta 5 desaturase activities were inhibited, the greatest reduction of delta 5 desaturase activity occurring with EPA. Incorporation of media fatty acids plus their desaturase and elongase products was highly dependent on medium composition with the homologues of delta 9 > delta 6 > delta 5. Supplementation of cellular media with 100 microM EPA for 10 days decreased membrane phosphatidylethanolamine arachidonic level from 13.2 to 8.9%. From these results we conclude that enterocyte membrane fatty composition and desaturase enzyme activity are regulated by both dietary fat intake and cell maturation. The clinical relevance of these observations on lipid dietary modification for the management of chronic inflammatory bowel disease is still uncertain but these observations suggest that the beneficial effects of EPA supplements on human ulcerative may be due to a reduction in enterocyte arachidonic content by down-regulation of delta 6 and delta 5 desaturase activity.

Keyword: inflammatory bowel disease

Short report: zinc sulphate supplementation corrects abnormal erythrocyte membrane long-chain fatty composition in patients with Crohn's .

Patients with Crohn's may become zinc-deficient and, in such patients, an altered metabolism of radiolabelled long-chain fatty acids has been reported. We have investigated the possible reversal by zinc supplementation of altered long-chain fatty profiles of red cells in Crohn's . Twenty patients with long-standing Crohn's in clinical remission received 200 mg of zinc sulphate daily for 6 weeks. Phospholipid fatty profiles of washed red cells were analysed before and after zinc treatment and compared to those of 20 unsupplemented healthy controls. Plasma zinc levels in Crohn's were 72 +/- 8 micrograms/dL before zinc treatment and increased to 114 +/- 10 micrograms/dl after the therapy. Prior to zinc supplementation, the percentage of , stearic and oleic acids was significantly higher in Crohn's , while linoleic, arachidonic and n-3 fatty acids were reduced in Crohn's compared to healthy controls. Zinc supplementation abolished these pre-treatment differences in red-cell long-chain fatty profiles but did not affect plasma fatty values. Further studies are needed to clarify whether these fatty changes can be related to the clinical course of the .

Keyword: inflammatory bowel disease

Docosahexaenoyl serotonin emerges as most potent inhibitor of IL-17 and CCL-20 released by blood mononuclear cells from a series of N-acyl serotonins identified in human intestinal tissue.

Fatty amides (FAAs), conjugates of fatty acids with ethanolamine, mono-amine neurotransmitters or amino acids are a class of molecules that display diverse functional roles in different cells and tissues. Recently we reported that one of the serotonin-fatty conjugates, docosahexaenoyl serotonin (DHA-5-HT), previously found in gut tissue of mouse and pig, attenuates the IL-23-IL-17 signaling axis in LPS-stimulated mice macrophages. However, its presence and effects in humans remained to be elucidated. Here, we report for the first time its identification in human intestinal (colon) tissue, along with a series of related N-acyl serotonins. Furthermore, we tested these fatty conjugates for their ability to inhibit the release of IL-17 and CCL-20 by stimulated human peripheral blood mononuclear cells (PBMCs). Serotonin conjugates with (PA-5-HT), stearic (SA-5-HT) and oleic (OA-5-HT) were detected in higher levels than arachidonoyl serotonin (AA-5-HT) and DHA-5-HT, while eicosapentaenoyl serotonin (EPA-5-HT) could not be quantified. Among these, DHA-5-HT was the most potent in inhibiting IL-17 and CCL-20, typical Th17 pro-inflammatory mediators, by Concanavalin A (ConA)-stimulated human PBMCs. These results underline the idea that DHA-5-HT is a gut-specific endogenously produced mediator with the capacity to modulate the IL-17/Th17 signaling response. Our findings may be of relevance in relation to intestinal inflammatory diseases like Crohn's disease and Ulcerative .Copyright © 2017. Published by Elsevier B.V.

Keyword: inflammatory bowel disease

[Chemically prepared fats and Crohn . A pilot study of the occurrence of trans-fatty acids in the subcutaneous tissue of Crohn patients in comparison with healthy controls as a parameter of long-term fat intake].

In a pilot study the fatty pattern of subcutaneous adipose tissue from 22 patients with Crohn's and 22 subjects of a healthy control group was analyzed using glass capillary gas-liquid chromatography. Among all fatty acids amounting to at least 1% peak area of the chromatograms, only trans-octadecenoate differed significantly (p less than 0.05) between both study groups, the mean value being 2.39 +/- 0.83% in patients with Crohn's and 1.96 +/- 0.46% in healthy controls. Also the mean value of trans-hexadecenoate was significantly (p less than 0.05) higher in the Crohn group (0.25 +/- 0.07%) than in the control group (0.21 +/- 0.06%). There was a strongly positive linear correlation (p less than 0.001) between the trans-hexadecenoate and trans-octadecenoate values for the Crohn patients but not for the controls. Our results demonstrate that patients with Crohn's as a group consume more trans-monoene fatty acids than healthy controls, thus providing evidence for a higher intake of chemically processed fats like margarine, shortenings, frying and cooking fats. In further studies which are necessary to examine Guthy's hypothesis the fatty composition of adipose tissue should be followed up as an ideal marker of long-term dietary compliance.

Keyword: inflammatory bowel disease

Comprehensive genetic study of fatty acids helps explain the role of noncoding inflammatory bowel disease associated SNPs and fatty metabolism in disease pathogenesis.

Fatty acids and their derivatives play an important role in inflammation. Diet and genetics influence fatty profiles. Abnormalities of fatty profiles have been observed in inflammatory bowel diseases (IBD), a group of complex diseases defined by chronic gastrointestinal inflammation. IBD associated fatty profile abnormalities were observed independently of nutritional status or disease activity, suggesting a common genetic background. However, no study so far has attempted to look for overlap between IBD loci and fatty associated loci or investigate the genetics of fatty profiles in IBD. To this end, we conducted a comprehensive genetic study of fatty profiles in IBD using iCHIP, a custom microarray platform designed for deep sequencing of immune-mediated disease associated loci. This study identifies 10 loci associated with fatty profiles in IBD. The most significant associations were a locus near CBS (p\u202f=\u202f7.62\u202f×\u202f10) and a locus in LRRK2 (p\u202f=\u202f1.4\u202f×\u202f10). Of note, this study replicates the FADS gene cluster locus, previously associated with both fatty profiles and IBD pathogenesis. Furthermore, we identify 18 carbon chain trans-fatty acids (p\u202f=\u202f1.12\u202f×\u202f10), total trans-fatty acids (p\u202f=\u202f4.49\u202f×\u202f10), (p\u202f=\u202f5.85\u202f×\u202f10) and arachidonic (p\u202f=\u202f8.58\u202f×\u202f10) as significantly associated with IBD pathogenesis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: inflammatory bowel disease

N-Acylethanolamine-hydrolyzing amidase inhibition increases colon N-palmitoylethanolamine levels and counteracts murine .

N-Palmitoylethanolamine or palmitoylethanolamide (PEA) is an anti-inflammatory compound that was recently shown to exert peroxisome proliferator-activated receptor-α-dependent beneficial effects on colon inflammation. The actions of PEA are terminated following hydrolysis by 2 enzymes: fatty amide hydrolase (FAAH), and the less-studied N-acylethanolamine-hydrolyzing amidase (NAAA). This study aims to investigate the effects of inhibiting the enzymes responsible for PEA hydrolysis in colon inflammation in order to propose a potential therapeutic target for inflammatory bowel diseases (IBDs). Two murine models of IBD were used to assess the effects of NAAA inhibition, FAAH inhibition, and PEA on macroscopic signs of colon inflammation, macrophage/neutrophil infiltration, and the expression of proinflammatory mediators in the colon, as well as on the -related systemic inflammation. NAAA inhibition increases PEA levels in the colon and reduces colon inflammation and systemic inflammation, similarly to PEA. FAAH inhibition, however, does not increase PEA levels in the colon and does not affect the macroscopic signs of colon inflammation or immune cell infiltration. This is the first report of an anti-inflammatory effect of a systemically administered NAAA inhibitor. Because NAAA is the enzyme responsible for the control of PEA levels in the colon, we put forth this enzyme as a potential therapeutic target in chronic inflammation in general and IBD in particular.© FASEB.

Keyword: inflammatory bowel disease

High-fat diet promotes experimental by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) tight junction regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes in a 2,4,6-trinitrobenzenesulfonic -induced experimental model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: inflammatory bowel disease

Preventive and curative effect of Pistacia lentiscus oil in experimental .

to investigate the anti-inflammatory effect of the Pistacia lentiscus oil in experimental model. was induced in male rats by instillation of 2,4,6-trinitrobenzenesulfonic (TNBS) in all groups. The experimental groups consisted of: 5 rats received Lentisc oil 2months before induction (preventive group), 5 rats received the oil on the day of induction (curative group) and 5 control rats. Lentisc oil was extracted from the ripe fruit of the plant by the cold press method and was analyzed by spectro-chromatography. Lentisc oil has been inserted with a standard diet at the dose of 30mg oil/100g of food/rat.The lentisc oil sample is composed mainly by Oleic (47.96%), (27.94%) and Linoleic (20.22%).There was a significant difference between control rats and treated rats with lentisc oil concerned body mass (p=0.009), bleeding index (p=0.005 and p=0.018) and diarrhea (p=0.012). Histological examination revealed a clear difference between the control and preventive groups with disappearance of erosion, decreased of cryptitis, irregular crypts and crypt loss in the preventive group. Curative group showed a significant decrease of ulceration, hyperplasia, cryptitis, irregular crypts and crypt loss compared to the control group. There was an attenuation of inflammation in the preventive group compared to the curative group without statistically significant.Lentisc oil administration could provide a protective effect on intestinal inflammation in rats induced by TNBS mainly when it is administered at a young age in preventive mode. This beneficial effect would involve a modification of arachidonic metabolism.Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Keyword: inflammatory bowel disease

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Trigonelline prevents high cholesterol and high fat diet induced hepatic lipid accumulation and lipo-toxicity in C57BL/6J mice, via restoration of hepatic autophagy.

Non-alcoholic fatty liver disease (NAFLD) is often linked with impaired hepatic autophagy. Here, we studied the alterations in hepatocellular autophagy by high cholesterol and high-fat diet (HC-HF) diet in C57BL/6J mice, and by (PA), in AML-12 and HepG2 cells. Further, we analysed role of Trigonelline (TG), a plant alkaloid, in preventing NAFLD, by modulating autophagy. For this, C57BL/6J mice were fed with Standard Chow (SC) or HC-HF diet, with and without TG for 16 weeks. In-vitro; AML-12\u202fcells and HepG2 cells, were exposed to PA with and without TG, for 24\u202fh. Cellular events related to autophagy, lipogenesis, and lipo-toxicity were studied. The HC-HF diet fed mice showed hepatic autophagy blockade, increased triglycerides and steatosis. PA exposure to AML-12\u202fcells and HepG2 cells induced impaired autophagy, ER stress, resulting in lipotoxicity. TG treatment in HC-HF fed mice, restored hepatic autophagy, and prevented steatosis. TG treated AML-12, and HepG2 cells exposed to PA showed autophagy restoration, and reduced lipotoxicity, however, these effects were diminished in Atg7-/- HepG2 cells, and in the presence of chloroquine. This study shows that HC-HF diet-induced impaired autophagy, and steatosis is prevented by TG, which attributes to its novel mechanism in treating NAFLD.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

BH3 mimetics derived from Bim-BH3 domain core region show PTP1B inhibitory activity.

A series of our previously described BH3 peptide mimetics derived from Bim-BH3 domain core region were found to exhibit weak to potent PTP1B binding affinity and inhibitory activities via target-based drug screening. Among these compounds, a 12-aa Bim-BH3 core sequence peptide conjugated to (SM-6) displayed good PTP1B binding affinity (K\u202f=\u202f8.38\u202fnmol/L), inhibitory activity (IC\u202f=\u202f1.20\u202fμmol/L) and selectivity against other PTPs (TCPTP, LAR, SHP-1 and SHP-2). Furthermore, SM-6 promoted HepG2 cell glucose uptake and inhibited the expression of PTP1B, indicating that SM-6 could improve the effect in the -resistant HepG2 cell model. These results may indicate a new direction for the application of BH3 peptide mimetics and promising PTP1B peptide inhibitors could be designed and developed based on SM-6.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to lipid metabolism and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and tolerance test (ITT) showed that compound C alleviated . Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: insulin resistance

Absence of Carbohydrate Response Element Binding Protein in Adipocytes Causes Systemic and Impairs Glucose Transport.

Lower adipose-ChREBP and de novo lipogenesis (DNL) are associated with in humans. Here, we generated adipose-specific ChREBP knockout (AdChREBP KO) mice with negligible sucrose-induced DNL in adipose tissue (AT). Chow-fed AdChREBP KO mice are resistant with impaired action in the liver, muscle, and\xa0AT and increased AT inflammation. HFD-fed AdChREBP KO mice are also more resistant\xa0than controls. Surprisingly, adipocytes lacking ChREBP display a cell-autonomous reduction in -stimulated glucose transport that is mediated by impaired Glut4 translocation and exocytosis, not lower Glut4 levels. AdChREBP KO mice have lower levels of esters of hydroxy stearic acids (PAHSAs) in serum, and AT. 9-PAHSA supplementation completely rescues their and AT inflammation. 9-PAHSA also normalizes impaired glucose transport and Glut4 exocytosis in ChREBP KO adipocytes. Thus, loss\xa0of adipose-ChREBP is sufficient to cause , potentially by regulating AT glucose transport and flux through specific lipogenic pathways.Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Keyword: insulin resistance

causes insulin resistance in granulosa cells via activation of JNK.

Obesity is a worldwide health problem with rising incidence and results in reproductive difficulties. Elevated saturated free fatty acids (FFAs) in obesity can cause insulin resistance (IR) in peripheral tissues. The high intra-follicular saturated FFAs may also account for IR in ovarian granulosa cells (GCs). In the present study, we investigated the relationship between saturated FFAs and IR in GCs by the use of (PA). We demonstrated that the glucose uptake in cultured GCs and lactate accumulation in the culture medium were stimulated by insulin, but the effects of insulin were attenuated by PA treatment. Besides, insulin-induced phosphorylation of Akt was reduced by PA in a dose and time-dependent manner. Furthermore, PA increased phosphorylation of JNK and JNK blockage rescued the phosphorylation of Akt which was down-regulated by PA. These findings highlighted the negative effect of PA on GCs metabolism and may partially account for the obesity-related reproductive disorders.

Keyword: insulin resistance

Scopoletin intervention in pancreatic endoplasmic reticulum stress induced by lipotoxicity.

Endoplasmic reticulum (ER), a dynamic organelle, plays an essential role in organizing the signaling pathways involved in cellular adaptation, resilience, and survival. Impairment in the functions of ER occurs in a variety of nutritive disorders including obesity and type 2 diabetes. Here, we hypothesize that (scopoletin) SPL, a coumarin, has the potential to alleviate ER stress induced in vitro and in vivo models by lipotoxicity. To test this hypothesis, the ability of SPL to restore the levels of proteins of ER stress was analyzed. Rat insulinoma 5f (RIN5f) cells and Sprague Dawley rats were the models used for this study. Groups of control and high-fat, high-fructose diet (HFFD)-fed rats were treated with either SPL or 4-phenylbutyric . Status of ER stress was enumerated by quantitative RT-PCR, Western blot, electron microscopic, and immunohistochemical studies. Proximal proteins of ER stress inositol requiring enzyme 1 (IRE1), protein kinase like endoplasmic reticulum kinase (PERK), and activating transcription factor 6 (ATF6) were reduced in the β-cells by SPL. The subsequent signaling proteins X-box binding protein 1, eukaryotic initiation factor2α, activating transcription factor 4, and C/EBP homologous protein were also suppressed in their expression levels when treated with SPL. IRE1, PERK signaling leads to c-Jun-N-terminal kinases phosphorylation, a kinase that interrupts signaling, which was also reverted upon scopoletin treatment. Finally, we confirm that SPL has the ability to suppress the stress proteins and limit pancreatic ER stress which might help in delaying the progression of .

Keyword: insulin resistance

Glucagon-like peptide-1 analog prevents obesity-related glomerulopathy by inhibiting excessive autophagy in podocytes.

To investigate the role of glucagon-like peptide-1 analog (GLP-1) in high-fat diet-induced obesity-related glomerulopathy (ORG). Male C57BL/6 mice fed a high-fat diet for 12 wk were treated with GLP-1 (200 μg/kg) or 0.9% saline for 4 wk. Fasting blood glucose and and the expression of podocin, nephrin, phosphoinositide 3-kinase (PI3K), glucose transporter type (Glut4), and microtubule-associated protein 1A/1B-light chain 3 (LC3) were assayed. Glomerular morphology and podocyte foot structure were evaluated by periodic -Schiff staining and electron microscopy. Podocytes were treated with 150 nM GLP-1 and incubated with 400 μM (PA) for 12 h. The effect on autophagy was assessed by podocyte-specific Glut4 siRNA. and autophagy were assayed by immunofluorescence and Western blotting. The high-fat diet resulted in weight gain, ectopic glomerular lipid accumulation, increased , and fusion of podophyte foot processes. The decreased translocation of Glut4 to the plasma membrane and excess autophagy seen in mice fed a high-fat diet and in PA-treated cultured podocytes were attenuated by GLP-1. Podocyte-specific Glut4 siRNA promoted autophagy, and rapamycin-enhanced autophagy worsened the podocyte injury caused by PA. Excess autophagy in podocytes was induced by inhibition of Glut4 translocation to the plasma membrane and was involved in the pathology of ORG. GLP-1 restored sensitivity and ameliorated renal injury by decreasing the level of autophagy.

Keyword: insulin resistance

Hydrogen Sulphide Treatment Increases Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells.

Studies have reported attenuation of (IR) by improving phosphorylation of the signalling pathway. However, the upstream molecular signalling pathway is still elusive. In this study, Western blot was used to evaluate the phosphorylation level of the signalling pathway and the AMPK pathway. 2-NBDG was used to evaluate glucose uptake. Ca imaging was used to assess change of intracellular Ca concentration. We found that NaHS enhanced the intracellular Ca concentration and glucose uptake and activated the signalling cascade in a (PA)-induced IR model in C2C12 cells. Furthermore, activation of the IRS1/PI3K/AKT pathway and glucose uptake were decreased when AMPK or CaMKKβ was inhibited. Our study also showed that the mitochondrial electron transport chain, ATP production, and intramitochondrial cAMP declined in the IR model but that this effect was reversed by NaHS, an effect that may be mediated by the Ca/CaMKK2/AMPK and PI3K/AKT pathways. Our data indicate that HS improves activation of the signalling cascade and glucose uptake via activation of the Ca/CaMKK2/AMPK pathway and mitochondrial metabolism in C2C12 cells. Furthermore, NaHS protects mitochondrial function and maintains normal ATP production by activating the cAMP system and the Ca/CaMKK2/AMPK and PI3K/ATK pathways.

Keyword: insulin resistance

GCN2 deficiency protects against high fat diet induced hepatic steatosis and in mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic fatty homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced liver dysfunction, hepatic steatosis and . In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and lipogenesis, and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), fatty synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting lipogenesis and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the liver may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Docosahexaenoic Reduces -Induced Endoplasmic Reticulum Stress in Pancreatic Β Cells.

Endoplasmic reticulum (ER) stress leads to peripheral and the progression of pancreatic beta cell failure in type 2 diabetes. Although ER stress plays an important role in the pathogenesis of diabetes, it is indispensable for cellular activity. Therefore, when assessing the pathological significance of ER stress, it is important to monitor and quantify ER stress levels. Here, we have established a novel system to monitor ER stress levels quickly and sensitively, and using this method, we have clarified the effect of differences in glucose concentration and various fatty acids on the ER of pancreatic β cells. First, we developed a cell system that secretes Gaussia luciferase in culture medium depending on the activation of the GRP78 promoter. This system could sensitively monitor ER stress levels that could not be detected with real-time RT-PCR and immunoblotting. This system revealed that hyperglycemia does not induce unfolded protein response (UPR) in a short period of time in MIN6 cells, a mouse pancreatic β cell line. Physiological concentrations of , a saturated fatty , induced ER stress quickly, while physiological concentrations of oleic , an unsaturated fatty , did not. Docosahexaenoic , an n-3 unsaturated fatty , inhibited -induced ER stress. In this study, we have established a system that can sensitively detect ER stress levels of living cells in a short period of time. This system can be used to monitor the state of the ER in living cells and lead to the investigation of the significance of physiological or pathological ER stress levels.

Keyword: insulin resistance

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of . In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: , atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: insulin resistance

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 diabetes.

Metabolic stress-induced low-grade chronic inflammation plays an important role in the development of and type 2 diabetes (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the obesity-induced metabolic stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: insulin resistance

Changes of the Fatty Profile in Erythrocyte Membranes of Patients following 6-Month Dietary Intervention Aimed at the Regression of Nonalcoholic Fatty Liver Disease (NAFLD).

Nonalcoholic fatty liver disease (NAFLD) is closely related to the metabolism disorders of fatty acids. The pathogenesis of the disease includes an increased concentration of FFA in blood, an increase in the biosynthesis of fatty acids, and disorders in the process of -oxidation.The aim of the study was to analyze the fatty acids in erythrocyte membranes among 55 patients with NAFLD who were subjected to a 6-month dietary intervention in order to reduce fatty liver.Basic anthropometric and biochemical measurements were performed. The profile of fatty acids was measured in the membranes of erythrocytes and analyzed by gas chromatography. The dietary compliance was evaluated using 72-diary questionnaires, anthropometric measurements.With the reduction of fatty liver (p<0.01), the patients\' biochemical and anthropometric parameters were significantly improved. A significant decrease in the concentration of alanine aminotransferase (p<0.01) and asparagine aminotransferase (p<0.01) was observed, along with a decrease in the amount of (p<0.05) and (p<0.05). Significant changes in terms of the fatty profile were observed among patients who followed the dietary intervention. There was a noticeable tendency in terms of the reduction (p<0.055) and a significant reduction of stearic (p<0.05). Significant changes in the profile of fatty acids were also associated with the reductionof palmitoleic (p<0.05) and oleic acids (p<0.05). Another statistically significant change observed was the increase in polyunsaturated fatty acids. In particular (p<0.01) the rise of eicosapentaenoic (p<0.055) and docosahexaenoic acids (p<0.55) was noted.The profile of fatty acids turned out to be a potential biomarker of the liver changes during NAFLD regression. Further research is needed to fully elucidate the usefulness and applicability of our findings in the management of NAFLD.

Keyword: insulin resistance

Mangiferin Improved Palmitate-Induced- by Promoting Free Fatty Metabolism in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved .

Elevated free fatty (FFA) is a key risk factor for (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high-fat diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA metabolism in HepG2 and C2C12 cells. The model was used to quantify PA-induced lipid accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased -stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA metabolism were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: insulin resistance

Taurine improves low-level inorganic arsenic-induced by activating PPARγ-mTORC2 signalling and inhibiting hepatic autophagy.

Inorganic arsenic (iAs) is reportedly associated with the increased incidence of type 2 diabetes in the population. Here, we found that iAs exposure significantly decreased the expression of glycolytic genes and glycogen content and increased gluconeogenesis gene levels in C57/BL6J mice. The expression of peroxisome proliferator-activated receptor γ (PPARγ), and mechanistic target of rapamycin complex 2 (mTORC2) were decreased in the livers of iAs-treated mice. Furthermore, in iAs-treated HepG2 cells, we found that PPARγ agonist rosiglitazone (RGS) increased the expression of mTORC2, inhibited autophagy, and improved glucose metabolism. mTORC2 agonist inhibited autophagy and improved glucose metabolism as well as the autophagosome formation inhibitor 3-methyladenine. Taurine, a natural compound, reversed impaired glucose metabolism and decreased expression of PPARγ and mTORC2 induced by iAs in mice liver and HepG2 cells. These data indicated that taurine administration could ameliorate iAs-induced through activating PPARγ-mTORC2 signalling and subsequently inhibiting hepatic autophagy.© 2018 Wiley Periodicals, Inc.

Keyword: insulin resistance

Good Fats versus Bad Fats: A Comparison of Fatty Acids in the Promotion of , Inflammation, and Obesity.

Recently, debate has erupted in both the scientific community and throughout the lay public around whether a low-fat or low-carbohydrate diet is better for weight loss. In other words, is it better to cut fat or cut carbohydrate for weight loss. However, going beyond this debate (fat versus carbohydrate), are questions around whether certain fatty acids are worse for promoting , inflammation, and obesity. The overall evidence in the literature suggests that medium-chain saturated fats (such as lauric , found in coconut oil) and monounsaturated fat (oleic , found in olive oil) are less likely to promote , inflammation, and fat storage compared to long-chain saturated fatty acids (such as stearic found in large quantities in butter, but particularly found in palm oil) especially when consumed on top of a diet moderate in refined carbohydrates. Compared to long-chain saturated fats, lauric and oleic have an increased fatty oxidation rate, are more likely to be burned for energy and less likely to be stored in adipose tissue, and thus promote increased energy expenditure. Omega-6 polyunsaturated fatty acids (PUFAs), such as linoleic , as found in vegetable oils may contribute to obesity, whereas omega-3 PUFA may be protective. Importantly, both olive oil as part of a Mediterranean diet, and omega-3 from fish and fish oil have been proven to reduce risk of cardiovascular (CV) events.

Keyword: insulin resistance

Free Fatty Acids: Circulating Contributors of Metabolic Syndrome.

Metabolic syndrome induces an increased cardiovascular morbidity and mortality. Most importantly, the prevalence of metabolic syndrome in adult population is expanding. Both clinical and preclinical studies indicate that increased Free Fatty Acids (FFAs) are involved in the pathogenesis of and subsequent development of metabolic syndrome. The relevance of FFAs in protecting and restoring tissue function is quite vast. The search to correlate the functional deterioration of the tissues within the cardiovascular system and increased plasma concentrations of FFAs has been reported. The importance of reduction in the consumption of dietary fatty acids along with the identification of dysregulated genes responsible for persistent increased FFAs uptake and mitochondrial β-oxidation has been increasingly recognized. This review discusses the current empirical understanding of the different types of fatty acids and their metabolism and functions both in physiological and pathophysiological conditions. We also discuss in detail about the molecular and pathophysiological basis of increased FFAs, which augments Cardiovascular Disease (CVD).Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Keyword: insulin resistance

Upregulation of miR-181a impairs lipid metabolism by targeting PPARα expression in nonalcoholic fatty liver disease.

Recent studies have reported elevated expression of miR-181a in patients with non-alcoholic fatty liver disease (NAFLD), suggesting that it may play an important role in liver lipid metabolism and . We aimed to investigate the effect of miR-181a in lipid metabolism and find new treatments for NAFLD. The expression level of miR-181a in NAFLD patient serum and a (PA)-induced NAFLD cell model was examined by Q-PCR. Oil red O staining and triglyceride assays were used to assess lipid accumulation in hepatocytes. Western blotting was used to detect the protein expression levels of peroxisome proliferator-activated receptor-α (PPARα) and the fatty β-oxidation-related genes. Direct interactions were validated by dual-luciferase reporter gene assays. MiR-181a expression was significantly upregulated in the serum of NAFLD patients and PA-induced hepatocytes. Inhibition of miR-181a expression resulted in the increased expression of PPARα and its downstream genes, and PA-induced lipid accumulation in hepatocytes was also inhibited. Upregulation of miR-181a resulted in the downregulation of its direct target PPARα and downstream gene expression of PPARα as well as aggravated lipid accumulation in hepatocytes. At the same time, the increased expression of PPARα can offset lipid accumulation in hepatocytes induced by miR-181a mimics. This study demonstrates that reducing the expression of miR-181a may improve lipid metabolism in NAFLD. The downregulation of miR-181a expression can be a therapeutic strategy for NAFLD by modulating its target PPARα.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Analysis of Fatty Esters of Hydroxyl Fatty in Selected Plant Food.

Metabolic syndrome, characterized by obesity, low-grade inflammation, , hyperglycemia, dyslipidemia and hypertension, is a major risk factor for cardiovascular mortality. Preclinical studies on recently discovered classes of lipids - fatty esters of hydroxy fatty acids (FAHFA) have revealed their anti-inflammatory and -sensitizing potential. The FAHFA levels are significantly decreased in -resistant individuals, their application exhibited anti-inflammatory effects and restoring the glucose- homeostasis. The aim of our research was to analyze the overall FAHFA composition in a common diet, as only a partial FAHFA composition has been revealed so far (only the PAHSA subclass was analyzed in a few foods). A new approach to the FAHFAs analysis includes nano-LC and post-column modifier followed by negative ion mass spectrometry, in order to obtain maximum sensitivity. Analysis of different foods - oat (whole grain, coarse flakes and fine flakes), apple, clementine, lemon, strawberry, blueberry, mango, kiwi, avocado, pineapple, banana, onion, garlic, cherry tomato, carrot, parsley root, pepper and radish - exhibited wide inter-food variation in the FAHFA profiles. Sixteen analyzed FAHFAs (, oleic, palmitoleic and stearic hydroxy-esters) showed microgram to low nanogram levels (0.165\xa0ng/g - 32\xa0μg/g FW), with the highest abundancy in oat, clementine, garlic and pineapple. Stearic hydroxy stearic (SAHSA) was the most abundant FAHFA, especially in the food with antioxidative, anti-inflammatory and beneficial metabolic effects. In contrary, the PAHSA - previously proven to have the strongest antihyperglycemic and -sensitizing effects, was not present in some foods (radish, avocado, mango, lemon, cherry tomato, kiwi). Our study proves the importance of overall FAHFA analysis in food (especially in a functional food), because of their potential metabolic benefits and possible future incorporation in special diets.

Keyword: insulin resistance

Fasting rapidly increases fatty oxidation in white adipose tissue of young broiler chickens.

Upregulating the fatty oxidation capacity of white adipose tissue in mice protects against diet-induced obesity, inflammation and . Part of this capacity results from induction of brown-like adipocytes within classical white depots, making it difficult to determine the oxidative contribution of the more abundant white adipocytes. Avian genomes lack a gene for uncoupling protein 1 and are devoid of brown adipose cells, making them a useful model in which to study white adipocyte metabolism in vivo. We recently reported that a brief (5\xa0hour) period of fasting significantly upregulated many genes involved in mitochondrial and peroxisomal fatty oxidation pathways in white adipose tissue of young broiler chickens. The objective of this study was to determine if the effects on gene expression manifested in increased rates of fatty oxidation. Abdominal adipose tissue was collected from 21\xa0day-old broiler chicks that were fasted for 3, 5 or 7\xa0hours or fed ad libitum (controls). Fatty oxidation was determined by measuring and summing CO production and C-labeled -soluble metabolites from the oxidation of [1-C] . Fasting induced a progressive increase in complete fatty oxidation and citrate synthase activity relative to controls. These results confirm that fatty oxidation in white adipose tissue is dynamically controlled by nutritional status. Identifying the underlying mechanism may provide new therapeutic targets through which to increase fatty oxidation in situ and protect against the detrimental effects of excess free fatty acids on adipocyte sensitivity.

Keyword: insulin resistance

Suppression of Rho-kinase 1 is responsible for regulation of the AMPK/SREBP-1c pathway in skeletal muscle cells exposed to palmitate.

Clinical and experimental data suggest that early therapy could reduce lipotoxicity in subjects and animal models with type 2 diabetes mellitus. However, the underlying mechanisms need to be clarified. Sterol regulatory element-binding protein 1c (SREBP-1c), which is negatively regulated by AMP-activated protein kinase (AMPK), plays a critical role in lipotoxicity and in skeletal muscle cells. Here, we investigated the effect and molecular mechanism of intervention on the AMPK/SREBP-1c pathway in skeletal muscle cells with chronic exposure to (PA).Male C57BL/6 mice were fed with a high-fat diet for 12\xa0weeks and were then treated with , AMPK inhibitor, or metformin. L6 myotubes incubated with (PA) were treated with or metformin. Dominant-negative AMPKα2 (DN-AMPKα2) lentivirus, AMPKα2 siRNA, or Rho-kinase 1 (ROCK1) siRNA were transfected into PA-treated L6 myotubes.We found that the ability of PA to stimulate SREBP-1c and inhibit AMPK was reversed by in L6 cells. Moreover, DN-AMPKα2 lentivirus and AMPKα2 siRNA were transfected into PA-treated L6 myotubes, and the decrease in SREBP-1c expression caused by was blocked by AMPK inhibition independent of the phosphatidylinositol-4,5-biphosphate-3-kinase (PI3K)/AKT pathway. The serine/threonine kinase Rho-kinase (ROCK) 1, a downstream effector of the small G protein RhoA, was activated by PA. Interestingly, knockdown of ROCK1 by siRNA blocked the downregulation of AMPK phosphorylation under PA-treated L6 myotubes, which indicated that ROCK1 mediated the effect of action on AMPK.Our study indicated that reduced lipotoxicity via ROCK1 and then improved AMPK/SREBP-1c signaling in skeletal muscle under PA-induced .

Keyword: insulin resistance

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of . The models were used to test the effect of 5-PAHSA on signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: insulin resistance

Erythropoietin ameliorates PA-induced through the IRS/AKT/FOXO1 and GSK-3β signaling pathway, and inhibits the inflammatory response in HepG2 cells.

Erythropoietin\xa0(EPO) contributes to in fat and muscle. In the present study, the role and mechanism of EPO in hepatic were investigated in HepG2 cells. Hepatic was induced by \xa0(PA) in the HepG2 cells, which were then treated with EPO\xa0(5 or 10\xa0U/ml) or specific phosphoinositide 3‑kinase\xa0(PI3K) inhibitors, wortmannin or LY294002. EPO treatment significantly increased glycogen levels and reduced the protein expression of phosphoenolpyruvate carboxykinase in the PA‑induced HepG2 cells. EPO also inhibited the serine phosphorylation of receptor substrate (IRS)‑1 (Ser307) and IRS‑2 (Ser473), and increased the protein expression levels of PI3K, phosphorylated (p)‑protein kinase B (AKT), p‑forkhead box O1 (FOXO1) and p‑glycogen synthase kinase 3\xa0(GSK‑3) β. In agreement with these result, the expression of p‑FOXO1\xa0(Ser256) and p‑GSK‑3β\xa0(Ser9), downstream molecules of AKT, were enhanced by EPO treatment (P<0.05). The specific PI3K inhibitors, LY294002 and wortmannin, markedly inhibited the EPO‑mediated increases in p‑AKT (Ser473), p‑FOXO1 (Ser256) and p‑GSK‑3β (Ser9) in the PA‑induced HepG2 cells (P<0.05). The gene expression levels of tumor necrosis factor‑α, interleukin‑1β and monocyte chemoattractant protein‑1, and the p‑c‑Jun N‑terminal kinase (JNK)/total‑JNK ratio were markedly suppressed by EPO treatment. These findings suggested that EPO treatment improved hepatic glucose metabolism, potentially through the IRS/AKT/FOXO1 and GSK‑3β signaling pathway, which may be associated with its inhibitory effect on the inflammation-associated response.

Keyword: insulin resistance

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac .

Patients with type 2 diabetes (T2D) and/or (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased -stimulated glucose and FA uptake, as well as loss of endosomal acidification and signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to obesity, diabetes and , edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic is unknown. The effects of GRE on lipid-induced hepatic using palmitate-exposed C3A liver cells and obese resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as receptor () and receptor substrate 1 and 2 ( and ), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic by improving sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: insulin resistance

[The nicotinic as an inhibitor of lipolysis in the phylogenetically early visceral cells; also blocks hydrolysis of triglycerides in the phylogenetically late adipocytes.]

It is valid to consider effect of nicotinic as an -mimetic one. The uniformity of biologic effect of exogenous nicotinic and endogenous permits to become aware that a) the hypo-lipidemic activity of , inhibition of lipolysis in phylogenetically late -dependent adipocytes and decreasing of content of unesterified fatty acids in blood plasma are considered as a basis of hypoglycemic effect of ; b) nicotinic similar to blocks lipolysis too but in hormoneindependent visceral fatty cells. The counter-insular effect is manifested in vivo by exogenous and endogenous saturated fatty by force of physical chemical characteristics. The biological role of consists in regulation of metabolism of fatty acids mainly unesterified fatty acids and in absorption of glucose by all -dependent cells. It is supposed that contractile cells (myocytes and cardiomyocytes) cumulate glycogen for implementing glucose as a substrate in synthesis in situ de novo of ῲ-9 oleic mono-saturated fatty . The initiates synthesis because mitochondria process this mono-saturated fatty to β-oxidation with the highest constant of reaction velocity. This is conditioned by physical chemical characteristics of oleic mono-saturated fatty , positioning of double bond in the chain of fatty ; this ensures maximal efficiency of ATP gaining. In phylogenesis, the low efficient alternative of metabolism of fatty acids is formed the earliest. At the later stages of phylogenesis the cells, under becoming of motion function, worked out more efficient alternative of ATP synthesis from exogenous ῲ-9 oleic monosaturated fatty . At the late stages of phylogenesis, under becoming of biological function of locomotion (motion at the expense of contraction of cross-striated skeletal myocytes) began to activate absorption of glucose by cells with subsequent synthesis of endogenous ῲ-9 oleic mono-saturated fatty out of it. The aphysiological effect of environment factors in the form of derangement of biological function of trophology (feeding), surplus of unesterified fatty acids in food serve as a main cause of such a high rate of functional derangement - syndrome in populations. Hence, there is no reason to call it diabetes mellitus type II.

Keyword: insulin resistance

Sumoylation of PPARγ contributes to vascular endothelium through stabilizing the PPARγ-NcoR complex.

Sumoylation of peroxisome proliferator-activated receptor\xa0γ (PPARγ) affects its stabilization, sublocalization, and transcriptional activity. However, it remains largely unknown whether PPARγ sumoylation inhibits the transactivation effect, leading to endothelium (IR). To test this possibility, human umbilical vascular endothelial cells (HUVECs) with a 90% confluence were randomly allocated to two batches. One batch was first pretreated with or without vitamin E for 24\u2009hr and the other infected with adenoviruses containing either PIAS1-shRNA (protein inhibitor of activated STAT1-short hairpin RNA) or scramble shRNA. Cells were suffered from high glucose and (PA) exposure for further 48\u2009hr. The levels of PPARγ, p-IKK, IKK, and NcoR (nuclear corepressors) were measured by western blot analysis. The interaction of IKK and PIAS1, as well as the PPARγ sumoylation, were examined by coimmunoprecipitation. The results showed that the exposure of high glucose and PA\xa0induced reactive oxygen species (ROS) production and IKK activation in HUVECs, promoting the interaction of IKK and PIAS1 and the sumoylation of PPARγ. However, vitamin E and PIAS1-shRNA partially decreased ROS production and IKK activation induced by high glucose and PA exposure. These data indicate that ROS-IKK-PIAS1 pathway mediates PPARγ sumoylation, leading to endothelium IR via stabilizing PPARγ-NcoR complex. These findings benefit understanding of regulatory networks of signaling, which might provide a potential target to prevent and cure IR-related diseases.© 2019 Wiley Periodicals, Inc.

Keyword: insulin resistance

Telmisartan protects against high glucose/high lipid-induced apoptosis and secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and . Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: insulin resistance

[Biological role of : transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of . 1. Phylogenetically, in vivo is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late blocks lipolysis only in phylogenetically late -dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: insulin resistance

Chronic Olanzapine Treatment Induces Disorders of Plasma Fatty Profile in Balb/c Mice: A Potential Mechanism for Olanzapine-Induced .

Atypical antipsychotics such as olanzapine cause metabolic side effects leading to obesity and . The underlying mechanisms remain elusive. In this study we investigated the effects of chronic treatment of olanzapine on the fatty composition of plasma in mice.Twenty 8-week female Balb/c mice were randomly assigned to two groups: the OLA group and the control group. After treatment with olanzapine (10 mg/kg/day) or vehicle intraperitoneally for 8 weeks, fasting glucose, levels and oral glucose tolerance test were determined. Effects on plasma fatty profile and plasma indices of D5 desaturase, D6 desaturase and SCD1 activity were also investigated.Chronic administration of olanzapine significantly elevated fasting glucose and levels, impaired glucose tolerance, but did not increase body weight. Total saturated fatty acids and n-6 polyunsaturated fatty acids were significantly increased and total monounsaturated fatty acids were significantly decreased, while total n-3 polyunsaturated fatty acids showed no prominent changes. Chronic olanzapine treatment significantly up-regulated D6 desaturase activity while down-regulating D5 desaturase activity. (C16:0), dihomo-γ-linolenic (C20:3n-6) and D6 desaturase were associated with an increase probability of , whereas nervonic (C24:1) and SCD1 were significantly associated with a lower probability.All results indicated that such drug-induced effects on fatty profile in plasma were relevant for the metabolic adverse effects associated with olanzapine and possibly other antipsychotics. Further studies are needed to investigate geneticand other mechanisms to explain how plasma fatty acids regulate glucose metabolism and affect the risk of .

Keyword: insulin resistance

-like growth factor binding protein 7 accelerates hepatic steatosis and in non-alcoholic fatty liver disease.

An association between increased -like growth factor binding protein-7 (IGFBP7) expression and in metabolic diseases has been reported. However, the role and molecular mechanism of IGFBP-7 in non-alcoholic fatty liver disease (NAFLD) remains largely unknown. Therefore, the potential function of IGFBP7 in the pathological progression of NAFLD was explored in this investigation. For in vivo experiments, an animal model of NAFLD was established in C57BL/6 mice by feeding a high-fat diet (HFD), and IGFBP7 was knocked down by injecting adeno-associated adenovirus (AAV)-mediated short-hairpin (sh)-IGFBP7 into the liver. We found that AAV-sh-IGFBP7 treatment significantly alleviated hepatocyte injury and inhibited hepatic lipid accumulation by reducing lipogenesis-associated gene expression. Furthermore, downregulation of IGFBP7 markedly ameliorated IR and restored impaired signalling by elevating the phosphorylation levels of IRS-1, Akt and GSK3β in HFD-treated mice. Similar results were also confirmed by an in vitro study in a (PA)-stimulated HepG2 cell model. In conclusion, our study demonstrates that IGFBP7 contributes to hepatic steatosis and in NAFLD development, which might serve as a novel therapeutic agent for the treatment of NAFLD.© 2019 John Wiley & Sons Australia, Ltd.

Keyword: insulin resistance

stimulates energy metabolism and inhibits /PI3K/AKT signaling in differentiated human neuroblastoma cells: The role of mTOR activation and mitochondrial ROS production.

The high consumption of saturated lipids has been largely associated with the increasing prevalence of metabolic diseases. In particular, saturated fatty acids such as (PA) have been implicated in the development of in peripheral tissues. However, how neurons develop in response to lipid overload is not fully understood. Here, we used cultured rat cortical neurons and differentiated human neuroblastoma cells to demonstrate that PA blocks -induced metabolic activation, inhibits the activation of the /PI3K/Akt pathway and activates mTOR kinase downstream of Akt. Despite the fact that fatty acids are not normally used as a significant source of fuel by neural cells, we also found that short-term neuronal exposure to PA reduces the NAD/NADH ratio, indicating that PA modifies the neuronal energy balance. Finally, inhibiting mitochondrial ROS production with mitoTEMPO prevented the deleterious effect of PA on signaling. This work provides novel evidence of the mechanisms behind saturated fatty -induced and its metabolic consequences on neuronal cells.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

Fatty Composition of Plasma Phosphatidylcholine Determines Body Fat Parameters in Subjects with Metabolic Syndrome-Related Traits.

This study examines the associations of fatty acids (FAs) in plasma phosphatidylcholine (PC) with the anthropometrical and biochemical characteristic of patients with metabolic syndrome (MetS)-related traits.We analyzed the FA profiles of PC in 300 persons with MetS-related traits (152\u2009M/148F, mean age 46.9\u2009±\u20099.0 years) and in 70 healthy controls of the same age using a balanced men/women ratio and gas-liquid chromatography. Multivariate linear regression analysis was performed to determine the coefficients of determination (R) using FA proportions of the mentioned proband characteristics.The FA composition of PC in patients with MetS traits was only associated with waist circumference (R\u2009=\u20090.27), waist-to-hip ratio (WHR; R\u2009=\u20090.41), body fat percentage (R\u2009=\u20090.62), and fat mass (R\u2009=\u20090.29). Positive associations were found for dihomo-γ-linolenic (DGLA), , stearic (SA), α-linolenic (ALA), and eicosapentaenoic acids, whereas negative associations were found for linoleic (LA), oleic, and docosapentaenoic acids. Palmitoleic (POA) was positively associated with waist circumference but negatively with fat percentage. In controls, significant associations were found for waist circumference (R\u2009=\u20090.51), WHR (R\u2009=\u20090.53), body fat percentage (R\u2009=\u20090.60), and fat mass (R\u2009=\u20090.34). DGLA and saturated FA (SFA) were positively associated, whereas docosahexaenoic, adrenic, and cis-vaccenic acids were negatively associated. The study group differed from controls as follows: lower concentrations of LA and total n-6 FA, higher indices of delta-9-desaturase and delta-6 desaturase activity and higher proportions of POA, SA, ALA, DGLA, and SFA.We found significant associations (R >0.25) of FA in plasma PC with adiposity in middle-aged persons with MetS-related traits, but not with metabolic indices.

Keyword: insulin resistance

Cooperative stimulation of atherogenesis by lipopolysaccharide and\xa0-rich high fat diet in low-density lipoprotein receptor-deficient mice.

Either lipopolysaccharide (LPS) or high-fat diet (HFD) enriched with saturated fatty (SFA) promotes atherosclerosis. In this study, we investigated the effect of LPS in combination with SFA-rich HFD on atherosclerosis and how LPS and SFA interact to stimulate inflammatory response in vascular endothelial cells.Low-density lipoprotein receptor-deficient (LDLR) mice were fed a low-fat diet (LFD), HFD with low (PA) (LP-HFD), or HFD with high PA (HP-HFD) for 20 weeks. During the last 12 weeks, half mice received LPS and half received PBS. After treatment, metabolic parameters and aortic atherosclerosis were analyzed. To understand the underlying mechanisms, human aortic endothelial cells (HAECs) were treated with LPS and/or PA and proinflammatory molecule expression was quantified.The metabolic study showed that LPS had no significant effect on cholesterol, triglycerides, free fatty acids, but increased and . Both LP-HFD and HP-HFD increased body weight and cholesterol while LP-HFD increased glucose and HP-HFD increased triglycerides, , and . Analysis of aortic atherosclerosis showed that HP-HFD was more effective than LP-HFD in inducing atherosclerosis and LPS in combination with HP-HFD increased atherosclerosis in the thoracic aorta, a less common site for atherosclerosis, as compared with LPS or HP-HFD. To understand the mechanisms, results showed that LPS and PA synergistically upregulated adhesion molecules and proinflammatory cytokines in HAECs.LPS and PA-rich HFD cooperatively increased atherogenesis in the thoracic aorta. The synergy between LPS and PA on proinflammatory molecules in HAECs may play an important role in atherogenesis.Published by Elsevier B.V.

Keyword: insulin resistance

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and lipid metabolism.

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet lipid metabolism in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high-fat diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. and β-cell function were then assessed using homeostasis model assessment of (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced lipid accumulation, reversed cell apoptosis, and enhanced glucose-stimulated secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced lipid deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell lipid accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: insulin resistance

Role of the saturated fatty palmitate in the interconnected hypothalamic control of energy homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body energy and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to , obesity, and type 2 diabetes mellitus. Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and energy homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: insulin resistance

Hot topic: Ceramide inhibits sensitivity in primary bovine adipocytes.

In nonruminants, the sphingolipid ceramide inhibits sensitivity by inactivating protein kinase B (AKT) within the -signaling pathway. We have established that ceramide accrual develops with impaired systemic action in ruminants during the transition from gestation to lactation, dietary supplementation, or controlled nutrient restriction. We hypothesized that ceramide promotes AKT inactivation and antagonizes sensitivity in primary bovine adipocytes. Stromal-vascular cells were grown from bovine adipose tissue explants and cultured in differentiation media. To modify ceramide supply, we treated differentiated adipocytes with (1) myriocin, an inhibitor of de novo ceramide synthesis, or (2) cell-permeable C2:0-ceramide. -stimulated AKT activation (i.e., phosphorylation) and 2-deoxy-D-[H]-glucose (2DOG) uptake were measured. Treatment of adipocytes with myriocin consistently decreased concentrations of ceramide, monohexosylceramide, and lactosylceramide. The -stimulated ratio of phosphorylated AKT to total AKT was increased with myriocin but decreased with C2:0-ceramide. Moreover, adipocyte -stimulated 2DOG uptake was decreased with C2:0-ceramide and increased with myriocin. We conclude that ceramide inhibits -stimulated glucose uptake by downregulating AKT activation in primary bovine adipocytes.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Anthocyanins ameliorate palmitate-induced inflammation and in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade inflammation leading to . Adipocyte differentiation inhibitors are expected to be effective in preventing obesity and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from metabolic stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, inflammation, and in 3T3-L1 adipocytes. ACN extract pretreatment reduces lipid accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces inflammation with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce pathway with levels higher than control cells, supporting an sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against obesity comorbidities, due to their protective effects against inflammation/ in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and conditions related to obesity.© 2019 John Wiley & Sons, Ltd.

Keyword: insulin resistance

Metabolic flexibility to lipid availability during exercise is enhanced in individuals with high sensitivity.

Metabolic flexibility to lipid (MetFlex-lip) is the capacity to adapt lipid oxidation to lipid availability. Hypothetically, impaired MetFlex-lip in skeletal muscle induces accumulation of lipid metabolites that interfere with signaling. Our aim was to compare MetFlex-lip during exercise in subjects with low (Low_IS) vs. high (High_IS) sensitivity. Twenty healthy men were designated as Low_IS or High_IS on the basis of the median of the homeostatic model assessment of index. Groups had similar age, body mass index, and maximum oxygen uptake (V̇o). Subjects cycled at 50% V̇o until expending 650 kcal. Adaptation in lipid oxidation was calculated as the drop in respiratory quotient (RQ) at the end of exercise vs. the maximum RQ (ΔRQ). Lipid availability was calculated as the increase in circulating nonesterified fatty acids (NEFA) at the end of exercise vs. the minimum NEFA (ΔNEFA). ΔRQ as a function of ΔNEFA was used to determine MetFlex-lip. On average, RQ and circulating NEFA changed similarly in both groups. However, ΔRQ correlated with ΔNEFA in High_IS ( r\u2009=\u2009-0.83, P < 0.01) but not in Low_IS ( r\u2009=\u2009-0.25, P = 0.48) subjects. Thus the slope of the ΔRQ vs. ΔNEFA relationship was steeper in High_IS vs. Low_IS subjects (-0.139 ± 0.03 vs. -0.025 ± 0.03 RQ·mmol·l, respectively; P < 0.05), with similar intercepts. We conclude that in subjects with High_IS lipid-to-carbohydrate oxidation ratio adapts to the increased circulating NEFA availability during exercise. Such MetFlex-lip appears impaired in subjects with Low_IS. Whether a cause-effect relationship exists between impaired MetFlex-lip and low sensitivity remains to be determined.

Keyword: insulin resistance

Inhibition of hepatocyte nuclear factor 1b induces hepatic steatosis through DPP4/NOX1-mediated regulation of superoxide.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disorder that is closely associated with and type 2 diabetes. Previous studies have suggested that hepatocyte nuclear factor 1b (HNF1b) ameliorates . However, the role of HNF1b in the regulation of lipid metabolism and hepatic steatosis remains poorly understood. We found that HNF1b expression was decreased in steatotic livers. We injected mice with lentivirus (LV) expressing HNF1b shRNA to generate mice with hepatic knockdown of HNF1b. We also injected high fat (HF) diet-induced obese and db/db diabetic mice with LV expressing HNF1b to overexpress HNF1b. Knockdown of HNF1b increased hepatic lipid contents and induced in mice and in hepatocytes. Knockdown of HNF1b worsened HF diet-induced increases in hepatic lipid contents, liver injury and in mice and PA-induced lipid accumulation and impaired signaling in hepatocytes. Moreover, overexpression of HNF1b alleviated HF diet-induced increases in hepatic lipid content and in mice. Knockdown of HNF1b increased expression of genes associated with lipogenensis and endoplasmic reticulum (ER) stress. DPP4 and NOX1 expression was increased by knockdown of HNF1b and HNF1b directly bound with the promoters of DPP4 and NOX1. Overexpression of DPP4 or NOX1 was associated with an increase in lipid droplets in hepatocytes and decreased expression of DPP4 or NOX1 suppressed the effects of knockdown of HNF1b knockdown on triglyceride (TG) formation and signaling. Knockdown of HNF1b increased superoxide level and decreased glutathione content, which was inhibited by downregulation of DPP4 and NOX1. N-acetylcysteine (NAC) suppressed HNF1b knockdown-induced ER stress, TG formation and . (PA) decreased HNF1b expression which was inhibited by NAC. Taken together, these studies demonstrate that HNF1b plays an essential role in controlling hepatic TG homeostasis and sensitivity by regulating DPP4/NOX1mediated generation of superoxide.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for obesity and diabetes.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: insulin resistance

Celastrol reverses (PA)-caused TLR4-MD2 activation-dependent via disrupting MD2-related cellular binding to PA.

Elevated plasma statured fatty acids (FFAs) cause TLR4/MD2 activation-dependent inflammation and tolerance, which account for the occurrence and development of obesity. It has been confirmed that statured (PA) (the most abundant FFA) could bind MD2 to cause cellular inflammation. The natural compound celastrol could improve obesity, which is suggested via inhibiting inflammation, yet the detailed mechanism for celastrol is still unclear. As celastrol is reported to directly target MD2, we thought disrupting the binding between FFAs and MD2 might be one of the ways for celastrol to inhibit FFAs-caused inflammation and . In this study, we found evidence to support our hypothesis: celastrol could reverse PA-caused TLR4/MD2 activation-dependent , as determined by glucose-lowering ability, cellular glucose uptake, action-related proteins and TLR4/MD2/NF-κB activation. Bioinformatics and cellular experiments showed that both celastrol and PA could bind MD2, and that celastrol could expel PA from cells. Finally, celastrol could reverse high fat diet caused hyperglycemia and obesity, and liver NF-kB activations. Taking together, we proved that celastrol could reverses PA-caused TLR4-MD2 activation-dependent via disrupting PA binding to MD2.© 2018 Wiley Periodicals, Inc.

Keyword: insulin resistance

Effects of selenium on apoptosis and abnormal amino metabolism induced by excess fatty in isolated rat hepatocytes.

Increased serum free fatty (FFA) occurs in subjects with non-alcoholic fatty liver disease (NAFLD) and also triggers oxidative stress, apoptosis, and . Selenium (Se) is an antioxidant agent. However, the effect of Se on NAFLD or diabetes is still unclear. We investigated the effect of Se on apoptosis and abnormal amino metabolism initiated by excess FFA in isolated rat hepatocytes.Primary hepatocytes from rats were isolated and exposed to excessive FFA (0.5 mM oleate/ 2:1) and 0.1 μM Se. Se protected primary hepatocytes against oxidative stress and apoptosis induced by excess FFA, but did not play a role on abnormal amino metabolism and initiated by FFA in isolated rat hepatocytes.Although Se had the capability of preventing the apoptosis initiated by ROS, failed to be reversed in hepatocytes exposed to FFA. This failure may be attributed to the limitation of Se in regulating branched chain amino acids abundance. This indicates that apoptosis and might be involved in different pathways when isolated hepatocytes were exposed to FFA and Se.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: insulin resistance

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of -stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty metabolism, function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Inhibition of by PGE1 via autophagy-dependent FGF21 pathway in diabetic nephropathy.

is a critical process in the initiation and progression of diabetic nephropathy (DN). Alprostadil (Prostaglandin E1, PGE1) had protective effects on renal function. However, it is unknown whether PGE1 inhibited in renal tubule epithelial cells via autophagy, which plays a protective role in DN against . was induced by (PA) in human HK-2 cells, shown as the decrease of -stimulated AKT phosphorylation, glucose transporter-4 (GLUT4), glucose uptake and enhanced phosphorylation of receptor substrate 1(IRS-1) at site serine 307 (pIRS-1ser307) and downregulated expression of IRS-1. Along with less abundance of p62, autophagy markers LC3B and Beclin-1 significantly increased in HK-2 cells exposed to PA. Such abnormal changes were significantly reversed by PGE1, which mimicked the role of autophagy gene 7 small interfering RNA (ATG7 siRNA). Furthermore, PGE1 promoted the protein expression of autophagy-related fibroblast growth factor-21 (FGF21), which alleviated . Results from western blotting and immunohistochemistry indicated that PGE1 remarkably restored autophagy, and the FGF21 expression in rat kidney of type 2 diabetes mellitus (T2DM). Collectively, we demonstrated the potential protection of PGE1 on in renal tubules via autophagy-dependent FGF21 pathway in preventing the progression of DN.

Keyword: insulin resistance

Comparison of Fatty Profiles in a Group of Female Patients with Chronic Kidney Diseases (CKD) and Metabolic Syndrome (MetS)⁻Similar Trends of Changes, Different Pathophysiology.

Fatty (FA) profiles in the plasma of patients with metabolic syndrome and chronic kidney disease (CKD) seem to be identical despite their different etiology (dietary mistakes vs. cachexia). The aim of this study was to compare both profiles and to highlight the differences that could influence the improvement of the treatment of patients in both groups. The study involved 73 women, including 24 patients with chronic kidney disease treated with haemodialysis, 19 patients with metabolic syndrome (MetS), and 30 healthy women in the control group. A total of 35 fatty acids and derivatives were identified and quantified by gas chromatography. Intensified elongation processes from C10:0 to C16:0 were noted in both groups (more intense in MetS), as well as an increased synthesis of arachidonic (C20:4n6), which was more intense in CKD. Significant correlations of oleic (C18:1n9), gamma linoleic (C18:3n6), and docosatetraenoate (C22:4n6) with parameters of CKD patients were observed. In the MetS group, auxiliary metabolic pathways of oleic were activated, which simultaneously inhibited the synthesis of eicosapentanoic (EPA) and docosahexaenoic (DHA) from alpha lipoic (ALA). On the other hand, in the group of female patients with CKD, the synthesis of EPA and DHA was intensified. Activation of the synthesis of oleic (C18: 1n9 ct) and trans-vaccinic (C18:1) is a protective mechanism in kidney diseases and especially in MetS due to the increased concentration of saturated fatty (SFA) in plasma. The cause of the increased amount of all FAs in plasma in the CKD group, especially in the case of (C16:0) and derivatives stearic (C18:0) acids, may be the decomposition of adipose tissue and the progressing devastation of the organism, whereas, in the MetS group, dietary intake seems to be the main reason for the increase in SFA. Moreover, in MetS, auxiliary metabolic pathways are activated for oleic , which cause the simultaneous inhibition of EPA and DHA synthesis from ALA, whereas, in the CKD group, we observe an increased synthesis of EPA and DHA. The higher increase of nervonic (C24:1) in CKD suggests a higher degree of demyelination and loss of axons.

Keyword: insulin resistance

The IRE1α-XBP1s pathway promotes -stimulated glucose uptake in adipocytes by increasing PPARγ activity.

The peroxisome proliferator-activated receptor-γ (PPARγ) improves whole-body sensitivity by regulating the adipogenic and metabolic functions of mature adipocytes. We have previously demonstrated that an active splice variant of X-box binding protein 1 (XBP1s) enhances PPARγ expression during adipogenesis. In this study, we investigated the role of XBP1s, particularly with respect to PPARγ, in the mechanisms underlying sensitivity in mature adipocytes. was able to stimulate XBP1s generation by activating inositol-requiring enzyme 1 (IRE1) α and was also able to increase its transcriptional activity by inducing nuclear translocation. XBP1s also upregulated the levels of phosphorylated IRS1 and AKT, demonstrating a positive feedback regulatory mechanism linking and XBP1s. XBP1s enhanced the expression of fibroblast growth factor 21 and, in turn, increased PPARγ activity, translocation of GLUT4 to the cell surface, and glucose uptake rate in adipocytes. In addition, XBP1s abolished palmitate-induced in adipocytes by increasing adiponectin secretion, repressing the secretion of pro-inflammatory adipokines such as leptin, monocyte chemoattractant protein 1, and tumor necrosis factor α, and decreasing fatty release. These findings provide a novel mechanism by which XBP1s stimulate sensitivity in adipocytes through fibroblast growth factor 21 induction and PPARγ activation.

Keyword: insulin resistance

A combination of plasma phospholipid fatty acids and its association with incidence of type 2 diabetes: The EPIC-InterAct case-cohort study.

Combinations of multiple fatty acids may influence cardiometabolic risk more than single fatty acids. The association of a combination of fatty acids with incident type 2 diabetes (T2D) has not been evaluated.We measured plasma phospholipid fatty acids by gas chromatography in 27,296 adults, including 12,132 incident cases of T2D, over the follow-up period between baseline (1991-1998) and 31 December 2007 in 8 European countries in EPIC-InterAct, a nested case-cohort study. The first principal component derived by principal component analysis of 27 individual fatty acids (mole percentage) was the main exposure (subsequently called the fatty pattern score [FA-pattern score]). The FA-pattern score was partly characterised by high concentrations of linoleic , stearic , odd-chain fatty acids, and very-long-chain saturated fatty acids and low concentrations of γ-linolenic , , and long-chain monounsaturated fatty acids, and it explained 16.1% of the overall variability of the 27 fatty acids. Based on country-specific Prentice-weighted Cox regression and random-effects meta-analysis, the FA-pattern score was associated with lower incident T2D. Comparing the top to the bottom fifth of the score, the hazard ratio of incident T2D was 0.23 (95% CI 0.19-0.29) adjusted for potential confounders and 0.37 (95% CI 0.27-0.50) further adjusted for metabolic risk factors. The association changed little after adjustment for individual fatty acids or fatty subclasses. In cross-sectional analyses relating the FA-pattern score to metabolic, genetic, and dietary factors, the FA-pattern score was inversely associated with adiposity, triglycerides, liver enzymes, C-reactive protein, a genetic score representing , and dietary intakes of soft drinks and alcohol and was positively associated with high-density-lipoprotein cholesterol and intakes of polyunsaturated fat, dietary fibre, and coffee (p < 0.05 each). Limitations include potential measurement error in the fatty acids and other model covariates and possible residual confounding.A combination of individual fatty acids, characterised by high concentrations of linoleic , odd-chain fatty acids, and very long-chain fatty acids, was associated with lower incidence of T2D. The specific fatty pattern may be influenced by metabolic, genetic, and dietary factors.

Keyword: insulin resistance

Pterostilbene reverses mediated in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

(IR) is known to precede onset of type 2 diabetes and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: insulin resistance

and Oleic : The Yin and Yang of Fatty Acids in Type 2 Diabetes Mellitus.

Increased plasma non-esterified fatty acids (NEFAs) link obesity with and type 2 diabetes mellitus (T2DM). However, in contrast to the saturated FA (SFA) , the monounsaturated FA (MUFA) oleic elicits beneficial effects on sensitivity, and the dietary :oleic ratio impacts diabetes risk in humans. Here we review recent mechanistic insights into the beneficial effects of oleic compared with on and T2DM, including its anti-inflammatory actions, and its capacity to inhibit endoplasmic reticulum (ER) stress, prevent attenuation of the signaling pathway, and improve β cell survival. Understanding the molecular mechanisms of the antidiabetic effects of oleic may contribute to understanding the benefits of this FA in the prevention or delay of T2DM.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

Intestinal epithelial cell caveolin 1 regulates fatty and lipoprotein cholesterol plasma levels.

Caveolae and their structural protein caveolin 1 (CAV1) have roles in cellular lipid processing and systemic lipid metabolism. Global deletion of CAV1 in mice results in and increases in atherogenic plasma lipids and cholesterol, but protects from diet-induced obesity and atherosclerosis. Despite the fundamental role of the intestinal epithelia in the regulation of dietary lipid processing and metabolism, the contributions of CAV1 to lipid metabolism in this tissue have never been directly investigated. In this study the cellular dynamics of intestinal Cav1 were visualized in zebrafish and the metabolic contributions of CAV1 were determined with mice lacking CAV1 in intestinal epithelial cells (CAV1). Live imaging of Cav1-GFP and fluorescently labeled caveolae cargos shows localization to the basolateral and lateral enterocyte plasma membrane (PM), suggesting Cav1 mediates transport between enterocytes and the submucosa. CAV1 mice are protected from the elevation in circulating fasted low-density lipoprotein (LDL) cholesterol associated with a high-fat diet (HFD), but have increased postprandial LDL cholesterol, total free fatty acids (FFAs), palmitoleic , and . The increase in circulating FAs in HFD CAV1 mice is mirrored by decreased hepatic FAs, suggesting a non-cell-autonomous role for intestinal epithelial cell CAV1 in promoting hepatic FA storage. In conclusion, CAV1 regulates circulating LDL cholesterol and several FA species via the basolateral PM of enterocytes. These results point to intestinal epithelial cell CAV1 as a potential therapeutic target to lower circulating FFAs and LDL cholesterol, as high levels are associated with development of type II diabetes and cardiovascular disease.© 2017. Published by The Company of Biologists Ltd.

Keyword: insulin resistance

Cardiomyocyte-specific knockout of endothelin receptor a attenuates obesity cardiomyopathy.

Endothelin (ET)-1 is implicated in the pathophysiology of cardiovascular diseases although its role in obesity anomalies has not been fully elucidated. This study was designed to examine the impact of ET-1 receptor A (ET) ablation on obesity-induced changes in cardiac geometry and contractile function, as well as the mechanisms involved with a focus on autophagy. Cardiomyocyte-specific ET receptor knockout (ETAKO) and WT mice were fed either low-fat (10% calorie from fat) or high-fat (45% calorie from fat) diet for 24\u202fweeks. Glucose tolerance test was examined to confirm . High-fat diet intake compromised myocardial geometry (enlarged left ventricular diameters in systole and diastole), morphology (cardiac hypertrophy, increased wall thickness and interstitial fibrosis), contractile function (reduced fractional shortening, ejection fraction and cardiomyocyte shortening) and intracellular Ca handling, the effect of which was significantly attenuated by ETAKO. TUNEL staining revealed overt apoptosis in high-fat-fed group, the effect was reverted by ETAKO. Western blot analysis noted that high-fat intake downregulated leptin receptor and PPARγ, signaling (elevated basal/dampened -stimulated phosphorylation of Akt and IRS1), phosphorylation of AMPK, ACC, upregulated GATA-4, ANP, NFATc3, PPARα, m-TOR/p70s6k signaling, which were attenuated by ETAKO with the exception of AMPK/ACC. Furthermore, high-fat intake suppressed cardiac autophagy, which was abrogated by ETAKO. In cultured murine cardiomyocytes, challenged mimicked high-fat diet-induced hypertrophic and autophagic responses, the effect of which were abolished by the ET receptor antagonist BQ123 or mTOR inhibitor rapamycin. These results suggest that inhibition of ET rescues high-fat intake-induced cardiac anomalies possibly through autophagy regulation.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Acne vulgaris: The metabolic syndrome of the pilosebaceous follicle.

Acne vulgaris is an epidemic inflammatory disease of the human sebaceous follicle and represents the most common skin disease affecting about 85% of adolescents in Westernized populations. Acne vulgaris is primarily a disease of wealthy countries and exhibits higher prevalence rates in developed compared with developing countries. No acne has been found in non-Westernized populations still living under Paleolithic dietary conditions constraining hyperglycemic carbohydrates, milk, and dairy products. The high prevalence rates of adolescent acne cannot be explained by the predominance of genetic factors but by the influence of a Western diet that overstimulates the key conductor of metabolism, the nutrient- and growth factor-sensitive kinase mTORC1. Increased mTORC1 activity has been detected in lesional skin and sebaceous glands of acne patients compared with acne-free controls. Increased mTORC1 signaling is a characteristic feature of , obesity, type 2 diabetes mellitus, cancer, and neurodegenerative diseases. Acne vulgaris is a family member of mTORC1-driven diseases of civilization and represents the MetS of the sebaceous follicle.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Individual fatty acids in erythrocyte membranes are associated with several features of the metabolic syndrome in obese children.

Obesity leads to the clustering of cardiovascular (CV) risk factors and the metabolic syndrome (MetS) also in children and is often accompanied by non-alcoholic fatty liver disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 fatty acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting (r\u2009=\u20090.287). Fatty Liver Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

Keyword: insulin resistance

Human umbilical cord-derived mesenchymal stem cells ameliorate by suppressing NLRP3 inflammasome-mediated inflammation in type 2 diabetes rats.

is one of the most common and important pathological features of type 2 diabetes (T2D). Recently, is increasingly considered to be associated with systemic chronic inflammation. Elevated levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in blood are predictive indicators of the development of T2D. Mesenchymal stem cell (MSC)-based therapies have been proven to have potential immunomodulation and anti-inflammatory properties through their paracrine effects; however, the mechanism for the anti-inflammatory effect of MSCs in enhancing sensitivity is still uncertain.In the present experiment, we used HepG2 cells, a human hepatoma cell line, and a MSC-HepG2 transwell culturing system to investigate the anti-inflammatory mechanism of human umbilical cord-derived MSCs (UC-MSCs) under (PA) and lipopolysaccharide (LPS)-induced in vitro. was confirmed by glycogen assay kit and glucose assay kit. Inflammatory factor release was detected by ELISA, gene expression was tested by quantitative real-time PCR, and signaling activation was determined by western blotting analysis. The changes of inflammatory factors and signaling protein were also tested in T2D rats injected with UC-MSCs.Treating HepG2 cells with PA-LPS caused NLRP3 inflammation activation, including overexpression of NLRP3 and caspase-1, and overproduction of IL-1β and IL-18 as well as TNF-α from HepG2 cells. The elevated levels of these inflammatory cytokines impaired receptor action and thereby prevented downstream signaling pathways, exacerbating in HepG2 cells. Importantly, UC-MSCs cocultured with HepG2 could effectively alleviate PA and LPS-induced by blocking the NLRP3 inflammasome activation and inflammatory agents. Furthermore, knockdown of NLRP3 or IL-1β partially improved PA and LPS-induced signaling impairments in the presence of UC-MSCs. Similarly, UC-MSC infusion significantly ameliorated hyperglycemia in T2D rats and decreased inflammatory activity, which resulted in improved sensitivity in target tissues.Our results indicated that UC-MSCs could attenuate and this regulation was correlated with their anti-inflammatory activity. Thus, MSCs might become a novel therapeutic strategy for and T2D in the near future.

Keyword: insulin resistance

Argirein alleviates vascular endothelial through suppressing the activation of Nox4-dependent O production in diabetic rats.

in endothelial cells contributes to the development of cardiovascular disease in type 2 diabetes mellitus (T2DM). Therefore, there are great potential clinical implications in developing pharmacological interventions targeting endothelial . Our previous studies indicated that argirein which was developed by combining rhein with L-arginine by a hydrogen bond, could substantially relieved stress related exacerbation of cardiac failure and alleviated cardiac dysfunction in T2DM, which was associated with suppressing NADPH oxidase activity. However, it is unclear whether argirein treatment attenuates the vascular lesion and dysfunction in T2DM and its underlying mechanisms.The rat aortic endothelial cells (RAECs) were used to treat with (PA), a most common saturated free fatty , which could induce . It was showed that argirein increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of IRS-1-ser307 and AKT-ser473, consequently resulting in the increase of the production of eNOS and NO in PA-induced RAECs. We further found that argirein blocked the Nox4-dependent superoxide (O) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vitro, argirein increased the release of endothelial NO to relieve the vasodilatory response to acetylcholine and , and restored the expression of Nox4 and pIRS-1-ser307 in the aorta endothelium of high-fat diet (HFD)-fed rats following an injection of streptozocin (STZ).These results suggested that argirein could improve endothelial which was attributed to inhibiting Nox4-dependent redox signaling in RAECs. These studies thus revealed the novel effect of argirein to prevent the vascular complication in T2DM.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Human Milk and Donkey Milk, Compared to Cow Milk, Reduce Inflammatory Mediators and Modulate Glucose and Lipid Metabolism, Acting on Mitochondrial Function and Oleylethanolamide Levels in Rat Skeletal Muscle.

Milk from various species differs in nutrient composition. In particular, human milk (HM) and donkey milk (DM) are characterized by a relative high level of triacylglycerol enriched in in sn-2 position. These dietary fats seem to exert beneficial nutritional properties through N-acylethanolamine tissue modulation. The aim of this study is to compare the effects of cow milk (CM), DM, and HM on inflammation and glucose and lipid metabolism, focusing on mitochondrial function, efficiency, and dynamics in skeletal muscle, which is the major determinant of resting metabolic rate. Moreover, we also evaluated the levels of endocannabinoids and N-acylethanolamines in liver and skeletal muscle, since tissue fatty profiles can be modulated by nutrient intervention. To this aim, rats were fed with CM, DM, or HM for 4 weeks. Then, glucose tolerance and were analyzed. Pro-inflammatory and anti-inflammatory cytokines were evaluated in serum and skeletal muscle. Skeletal muscle was also processed to estimate mitochondrial function, efficiency, and dynamics, oxidative stress, and antioxidant/detoxifying enzyme activities. Fatty profiles, endocannabinoids, and N-acylethanolamine congeners were determined in liver and skeletal muscle tissue. We demonstrated that DM or HM administration reducing inflammation status, improves glucose disposal and and reduces lipid accumulation in skeletal muscle. Moreover, HM or DM administration increases redox status, and mitochondrial uncoupling, affecting mitochondrial dynamics in the skeletal muscle. Interestingly, HM and DM supplementation increase liver and muscle levels of the N-oleoylethanolamine (OEA), a key regulator of lipid metabolism and inflammation. HM and DM have a healthy nutritional effect, acting on inflammatory factors and glucose and lipid metabolism. This beneficial effect is associated to a modulation of mitochondrial function, efficiency, and dynamics and to an increase of OEA levels in skeletal muscle.

Keyword: insulin resistance

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

(IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Cyanidin-3-O-glucoside ameliorates palmitate-induced by modulating IRS-1 phosphorylation and release of endothelial derived vasoactive factors.

Increased plasma levels of free fatty acids, including (PA), cause in endothelium characterized by a decreased synthesis of -mediated vasodilator nitric oxide (NO), and by an increased production of the vasoconstrictor protein, endothelin-1. Several in vitro and in vivo studies suggest that anthocyanins, natural phenols commonly present in food and vegetables from Mediterranean Diet, exert significant cardiovascular health-promoting activities. These effects are possibly mediated by a positive regulation of the transcription factor Nrf2 and activation of cellular antioxidant and cytoprotective genes. The present study examined, at a molecular level, the effects of cyanidin-3-O-glucoside (C3G), a widely distributed anthocyanin, on PA-induced endothelial dysfunction and in human umbilical vein endothelial cells (HUVECs). Our results indicate that C3G pretreatment effectively reverses the effects of PA on PI3K/Akt axis, and restores eNOS expression and NO release, altered by PA. We observed that these effects were exerted by changes on the phosphorylation of IRS-1 on specific serine and tyrosine residues modulated by PA through the modulation of JNK and IKK activity. Furthermore, silencing Nrf2 transcripts demonstrated that the protective effects of C3G are directly related to the activation of Nrf2.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

triggers inflammatory responses in N42 cultured hypothalamic cells partially via ceramide synthesis but not via TLR4.

A high-fat diet induces hypothalamic inflammation in rodents which, in turn, contributes to the development of obesity by eliciting both and leptin . However, the mechanism by which long-chain saturated fatty acids trigger inflammation is still contentious. To elucidate this mechanism, the effect of fatty acids on the expression of the pro-inflammatory cytokines IL-6 and TNFα was investigated in the mHypoE-N42 hypothalamic cell line (N42). N42 cells were treated with lauric (LA) and (PA). PA challenge was carried out in the presence of either a TLR4 inhibitor, a ceramide synthesis inhibitor (L-cycloserine), oleic (OA) or eicosapentaenoic (EPA). Intracellular ceramide accumulation was quantified using LC-ESI-MS/MS. PA but not LA upregulated IL-6 and TNFα. L-cycloserine, OA and EPA all counteracted PA-induced intracellular ceramide accumulation leading to a downregulation of IL-6 and TNFα. However, a TLR4 inhibitor failed to inhibit PA-induced upregulation of pro-inflammatory cytokines. In conclusion, PA induced the expression of IL-6 and TNFα in N42 neuronal cells independently of TLR4 but, partially, via ceramide synthesis with OA and EPA being anti-inflammatory by decreasing PA-induced intracellular ceramide build-up. Thus, ceramide accumulation represents one on the mechanisms by which PA induces inflammation in neurons.

Keyword: insulin resistance

[ is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. is a pathology associated primarily with FA and secondarily with glucose.

Keyword: insulin resistance

The Haematococcus pluvialis extract enriched by bioaccumulation process with Mg(II) ions improves in equine adipose-derived stromal cells (EqASCs).

(IR) is one of the characteristic features of equine metabolic syndrome (EMS). Presently, the only therapies of choice are caloric restrictions combined with mineral supplementation, which might improve sensitivity. In this study we investigated the effect of Haematococcus pluvialis algae water extract enriched in bioaccumulation process in magnesium ions (Hp_Mg(II)) on equine adipose derived mesenchymal stromal stem cells, in which was induced by (IR-EqASCs). For this purpose, chemical characterization of H. pluvialis was performed with special emphasis on the analysis of minerals composition, total phenolic and carotenoids contents, as well as scavenging activity. To examine the influence of H. pluvialis extract on IR-EqASCs, various methods of molecular biology and microscopic observations (i.e., immunofluorescence staining, SEM, gene expression by RT-qPCR, proliferative and metabolic cells activity analysis) were applied to investigate in vitro viability, oxidative stress markers and apoptosis-related factor accumulation, along with -related genes expression. Obtained results show, that Hp_Mg(II) significantly improves proliferative and metabolic activity of IR-EqASCs, shortens their population doubling time, improves their clonogenic potential and reduces expression of apoptosis related genes. Moreover, anti-oxidative effect of extract was presented.Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: insulin resistance

Azoramide improves mitochondrial dysfunction in palmitate-induced resistant H9c2 cells.

Azoramide is identified as a new compound with the dual properties for the improvement of ER-folding capacity in various cells as well as for the treatment of T2DM. Although the effect of azoramide in glucose-homeostasis in mammalians is not known very well, a limited number of experimental studies showed that it could improve the sensitivity in genetically obese mice. Therefore, here, we aimed to investigate the direct effect of azoramide on signaling in -resistant (IR) cardiomyocytes using IR-modelled ventricular cardiomyocytes. This model was established in H9c2 cells using incubation (50-μM for 24-h). The development of IR in cells was verified by monitoring the cellular 2-DG6P uptake assays in these treated cells. The 2-DG6P uptake was 50% less in the IR-cells compared to the control cells, while azoramide treatment (20-μM for 48-h) could prevent fully that decrease. In addition, azoramide treatment markedly preserved the IR-induced less ATP production and high-ROS production in these IR-cells. Furthermore, this treatment prevented the functional changes in mitochondria characterized by depolarized mitochondrial membrane potential and mitochondrial fusion or fusion-related protein levels as well as cellular ATP level. Moreover, this treatment provided marked protection against IR-associated changes in the signaling pathway in cells, including recovery in the phosphorylation of IRS1 and Akt as well as the protein level of GLUT4 and Akt. Our present results, for the first time, demonstrated that azoramide plays an important protective role in IR-cardiomyocytes, at most, protective action on mitochondria. Therefore, one can suggest that azoramide, as a novel regulator, can provide direct cardioprotection in the IR-heart, at most, via affecting mitochondria and can be a good candidate as a new drug for the treatment of IR-associated cardiovascular disorders in mammalians with systemic IR.

Keyword: insulin resistance

Sphingolipid changes do not underlie fatty -evoked GLUT4 nor inflammation signals in muscle cells.

Ceramides contribute to obesity-linked and inflammation in vivo, but whether this is a cell-autonomous phenomenon is debated, particularly in muscle, which dictates whole-body glucose uptake. We comprehensively analyzed lipid species produced in response to fatty acids and examined the consequence to and pro-inflammatory pathways. L6 myotubes were incubated with BSA-adsorbed palmitate or palmitoleate in the presence of myriocin, fenretinide, or fumonisin B1. Lipid species were determined by lipidomic analysis. sensitivity was scored by Akt phosphorylation and glucose transporter 4 (GLUT4) translocation, while pro-inflammatory indices were estimated by IκBα degradation and cytokine expression. Palmitate, but not palmitoleate, had mild effects on Akt phosphorylation but significantly inhibited -stimulated GLUT4 translocation and increased expression of pro-inflammatory cytokines and Ceramides, hexosylceramides, and sphingosine-1-phosphate significantly heightened by palmitate correlated negatively with sensitivity and positively with pro-inflammatory indices. Inhibition of sphingolipid pathways led to marked changes in cellular lipids, but did not prevent palmitate-induced impairment of -stimulated GLUT4 translocation, suggesting that palmitate-induced accumulation of deleterious lipids and are correlated but independent events in myotubes. We propose that muscle cell-endogenous ceramide production does not evoke and that deleterious effects of ceramides in vivo may arise through ancillary cell communication.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: insulin resistance

Rare ginsenosides ameliorate lipid overload-induced myocardial via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 diabetes. However, their pharmacological studies regarding myocardial fuel metabolism and signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel metabolism and signaling.We used (PA) to treat H9c2 cells in vitro and high fat diet (HFD) to mice to induce in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated -mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial sensitivity under conditions of lipid overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: insulin resistance

Inhibition of central de novo ceramide synthesis restores signaling in hypothalamus and enhances β-cell function of obese Zucker rats.

Hypothalamic lipotoxicity has been shown to induce central and dysregulation of glucose homeostasis; nevertheless, elucidation of the regulatory mechanisms remains incomplete. Here, we aimed to determine the role of de novo ceramide synthesis in hypothalamus on the onset of central and the dysregulation of glucose homeostasis induced by obesity.Hypothalamic GT1-7 neuronal cells were treated with palmitate. De novo ceramide synthesis was inhibited either by pharmacological (myriocin) or molecular (si-Serine Palmitoyl Transferase 2, siSPT2) approaches. Obese Zucker rats (OZR) were intracerebroventricularly infused with myriocin to inhibit de novo ceramide synthesis. was determined by quantification of Akt phosphorylation. Ceramide levels were quantified either by a radioactive kinase assay or by mass spectrometry analysis. Glucose homeostasis were evaluated in myriocin-treated OZR. Basal and glucose-stimulated parasympathetic tonus was recorded in OZR. secretion from islets and β-cell mass was also determined.We show that palmitate impaired signaling and increased ceramide levels in hypothalamic neuronal GT1-7 cells. In addition, the use of deuterated demonstrated that palmitate activated several enzymes of the de novo ceramide synthesis pathway in hypothalamic cells. Importantly, myriocin and siSPT2 treatment restored signaling in palmitate-treated GT1-7 cells. Protein kinase C (PKC) inhibitor or a dominant-negative PKCζ also counteracted palmitate-induced . Interestingly, attenuating the increase in levels of hypothalamic ceramides with intracerebroventricular infusion of myriocin in OZR improved their hypothalamic -sensitivity. Importantly, central myriocin treatment partially restored glucose tolerance in OZR. This latter effect is related to the restoration of glucose-stimulated secretion and an increase in β-cell mass of OZR. Electrophysiological recordings also showed an improvement of glucose-stimulated parasympathetic nerve activity in OZR centrally treated with myriocin.Our results highlight a key role of hypothalamic de novo ceramide synthesis in central installation and glucose homeostasis dysregulation associated with obesity.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: insulin resistance

[The person in philogenesis is not (omnivores), but the herbivores with the carnivores past and the fuzzy future. Biological function of trophology (nutrition) in ontogenesis.]

According to the phylogenetic theory of general pathology, seven biological functions have been formed over billions of years. 1. biological function of trophology, nutrition; 2. homeostasis function; 3. biological function of endoecology; 4. function of adaptation; 5. function of the continuation of the species; 6. function of locomotion and 7. cognitive biological function, including intelligence. Millions of years in life consistently in the waters of several oceans, all the ancestors of man were carnivorous (Carnivores), fish-eating mammals. When the ocean retreated and the carnivorous (fish-eating) were on land, each individual privatized a "piece" of the ocean. Animals transformed it ito a pool of intercellular medium in vivo. The biological role of the late in the phylogeny of is the formation of new biological functions in vivo. The action of has transformed the carnivorous (fish-eating) ocean into herbivorous (Herbivores) species on land. There was it by synthesis in vivo from exogenous glucose of fatty acids (FA). Regulatory action of was the directed conversion of exogenous glucose into ω-6 C18: 1 cis-oleic FA. late in phylogeny expressed the synthesis of new, conjugated enzymes: it is palmitoyl-CoA-elongase and stearyl-CoAdesaturase. Two enzymes synthesized FAs along the way: synthesized in situ de novo, from exogenous glucose, C16: 0 → C18: 0 stearic → ω-6 C18: 1 cis-oleic without accumulation of stearic FA. is not converted into an oleic FA exogenous from carnivorous food. On land, the action of transformed the species Homo sapiens, into a herbivore, but with carnivorous, fish-eating, past. The idea of a person as omnivorous (Omnivor) - nonsense; such forms of nature did not form. Violation of the function of nutrition, the biological reaction of exotrophy (external nutrition), is the etiological and pathogenetic basis of the seven metabolic pandemics, the diseases of civilization. 1. Atherosclerosis and atheromatosis; 2. metabolic arterial hypertension; 3. metabolic syndrome; 4. obesity; 5. syndrome of ; 6. non-alcoholic fatty liver disease and 7. endogenous hyperuricemia. The primary prevention of metabolic pandemics in the biological function of nutrition, in the biological reactions of exo-and endotrophy, will allow us to understand the theoretical bases and implementation of preventive actions that will determine the characteristics of nutrition in the future.

Keyword: insulin resistance

Chitosan Oligosaccharides Improve Glucolipid Metabolism Disorder in Liver by Suppression of Obesity-Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid metabolism disorder using (PA)-induced HepG2 cells and high-fat diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated lipid accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid metabolism-related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid metabolism disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid metabolism-related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid metabolism disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid metabolism-related diseases.

Keyword: insulin resistance

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and .

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic . The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and .By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: insulin resistance

A prospective and longitudinal study of plasma phospholipid saturated fatty profile in relation to cardiometabolic biomarkers and the risk of gestational diabetes.

Data on saturated fatty acids (SFAs) in relation to metabolic function and glucose homeostasis remain controversial. Such data are lacking among pregnant women.We prospectively investigated objectively measured individual and subclasses of plasma phospholipid SFAs throughout pregnancy in relation to cardiometabolic markers and gestational diabetes mellitus (GDM) risk.Within the National Institute of Child Health and Human Development Fetal Growth Studies-Singleton Cohort of 2802 singleton pregnancies, 107 GDM cases were ascertained via medical record review and matched to 214 non-GDM controls on age, race/ethnicity, and gestational week (GW) at blood collection. Individual plasma phospholipid SFA concentrations were repeatedly measured throughout pregnancy at GWs 10-14, 15-26, 23-31, and 33-39 and also grouped into subclasses of even- or odd-chain SFAs.From GW 10, even-chain SFA concentrations were significantly higher among women who later developed GDM, whereas odd-chain SFAs were significantly lower among GDM cases compared with controls. At GWs 10-14, the SFA (16:0) was positively associated with impaired and cardiometabolic markers and the risk of GDM [adjusted OR comparing the highest with the lowest quartile (aORQ4-Q1): 4.76; 95% CI: 1.72, 13.10; P-trend\xa0=\xa00.001]. In contrast, odd-chain SFAs were inversely related to the previously mentioned markers and GDM risk [aORQ4-Q1 for pentadecanoic (15:0): 0.32; 95% CI: 0.11, 0.92; P-trend\xa0=\xa00.025; for heptadecanoic (17:0): 0.20; 95% CI: 0.07, 0.58; P-trend\xa0=\xa00.003]. Women with high (median or greater) even-chain SFA concentrations and low (less than median) odd-chain SFAs had a 9.43-fold (95%: CI 3.26-, 27.30-fold) increased risk compared with women with low even-chain and high odd-chain SFA concentrations. Similar results were observed at GWs 15-26.The study provided one of the first lines of evidence suggesting that circulating concentrations of SFAs varying by SFA chain length, as early as GWs 10-14, were significantly and differentially associated with subsequent risk of GDM. Our findings highlight the importance of assessing objectively measured, individual, and subclasses of SFAs to investigate their distinct biological and pathophysiologic roles in glucose homeostasis and cardiometabolic outcomes. This study was registered at www.clinicaltrials.gov as .

Keyword: insulin resistance

Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with metabolic syndrome.

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an energy-restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with metabolic syndrome.The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of metabolic syndrome.After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk energy-restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Oleuropein improves in skeletal muscle by promoting the translocation of GLUT4.

As the beneficial effects of the Mediterranean diet on human health are well established, the phenolic compounds in olive oil have been gaining interest. Oleuropein, a major phenolic compound in olives, is known to reduce the blood glucose levels in alloxan-induced diabetic rats and rabbits, however, its effect on type 2 diabetes caused by obesity is not clear. The purpose of this study is clarifying the effect of oleuropein on the glucose tolerance in skeletal muscle under the condition of lipotoxicity caused by type 2 diabetes. Oleuropein enhanced glucose uptake in C2C12 cells without . Translocation of glucose transporter 4 (GLUT4) into the cell membrane was promoted by activation of adenosine monophosphate-activated protein kinase (AMPK) but not protein kinase B (Akt). Physiological concentration of oleuropein (10\xa0µM) was sufficient to express beneficial effects on C2C12 cells. Oleuropein prevented -induced myocellular . Furthermore, in gastrocnemius muscles of mice fed a high fat diet, oleuropein also induced the GLUT4 localization into cell membrane. These results suggest the possibility of oleuropein to be effective for type 2 diabetes by reducing in skeletal muscles.

Keyword: insulin resistance

Postprandial Saturated Fatty Acids Increase the Risk of Type 2 Diabetes: A Cohort Study in a Chinese Population.

Experimental evidence suggests saturated fatty acids (SFAs) are associated with , but results from epidemiological studies on fasting SFAs-diabetes risk are inconsistent.We investigated SFA (fasting and 2-hour postprandial) profiles and diabetes risk.A total of 8940 participants were recruited for the Harbin People\'s Health Study in 2008. Serum SFAs (fasting and 2-hour postprandial) at baseline in Chinese men and women without diabetes were profiled, and type 2 diabetes was ascertained using World Health Organization criteria after 4 to 7 years of follow-up.Associations between 2-hour postprandial SFA (2h-SFA) and diabetes.At baseline, incident cases of diabetes were older with a higher body mass index and waist circumference. After a mean follow-up of 6.7 years, 658 incident cases of diabetes occurred. After propensity score computation and inverse probability of treatment weighting (IPTW) estimation, fasting SFAs were unrelated to diabetes risk but IPTW-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the highest tertile of 2-hour postprandial stearic (2h-SA), 2-hour postprandial (2h-PA), and 2h-SFA for diabetes risk were 2.50 (2.08 to 3.16), 1.56 (1.23 to 2.02), and 1.70 (1.34 to 2.17), respectively (P-trend < 0.0001). Similarly, 2h-SA/fasting SA, 2h-PA/fasting PA, and 2h-SFA/fasting SFA ratios [IPTW-adjusted OR (95% CI): 2.94 (2.39 to 3.58), 2.31 (1.80 to 2.93), and 2.42 (1.91 to 3.11), respectively; P-trend < 0.0001] predicted the diabetes risk.Higher serum 2h-SFA (but not fasting SFA) independently predicted diabetes risk.

Keyword: insulin resistance

Hepatic fatty biosynthesis in KK-A mice is modulated by administration of persimmon peel extract: A DNA microarray study.

Previously, we showed that the intake of a persimmon peel (PP) extract altered hepatic gene expression associated with the signaling pathway and enhanced tyrosine phosphorylation of receptors in nonobese type 2 diabetic Goto-Kakizaki rats. Our objective was to evaluate the effect of fat-soluble PP extract on obese type 2 diabetic KK-A mice with .KK-A mice were fed a diet mixed with 0.1% of the extract for 8\xa0weeks. The total ketone body levels in the plasma of PP extract-fed mice were significantly lower than those in the normal diet-fed mice. Hepatic nonesterified content was higher in the PP extract-fed mice than in normal diet-fed mice. The hepatic gene expression profiles of the treated mice indicated upregulation of fatty synthesis and downregulation of inflammation-associated genes, predicting SREBP-1c and PPARγ activation.These results suggest that the PP extract enhances hepatic fatty synthesis via SREBP-1c and PPARγ, as well as anti-inflammatory activity in KK-A mice.

Keyword: insulin resistance

feeding increases ceramide supply in association with increased milk yield, circulating nonesterified fatty acids, and adipose tissue responsiveness to a glucose challenge.

Reduced action is a key adaptation that facilitates glucose partitioning to the mammary gland for milk synthesis and enhances adipose tissue lipolysis during early lactation. The progressive recovery of sensitivity as cows advance toward late lactation is accompanied by reductions in circulating nonesterified fatty acids (NEFA) and milk yield. Because can promote in monogastrics through sphingolipid ceramide-dependent mechanisms, (C16:0) feeding may enhance milk production by restoring homeorhetic responses. We hypothesized that feeding C16:0 to mid-lactation cows would enhance ceramide supply and ceramide would be positively associated with milk yield. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows were randomly assigned to a sorghum silage-based diet containing no supplemental fat (control; n=10; 138±45 d in milk) or C16:0 at 4% of ration dry matter (PALM; 98% C16:0; n=10; 136±44 d in milk). Blood and milk were collected at routine intervals. Liver and skeletal muscle tissue were biopsied at d 47 of treatment. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. The plasma and tissue concentrations of ceramide and glycosylated ceramide were determined using liquid chromatography coupled with tandem mass spectrometry. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield, energy-corrected milk, and milk fat yield. The most abundant plasma and tissue sphingolipids detected were C24:0-ceramide, C24:0-monohexosylceramide (GlcCer), and C16:0-lactosylceramide. Plasma concentrations of total ceramide and GlcCer decreased as lactation advanced, and ceramide and GlcCer were elevated in cows fed PALM. feeding increased hepatic ceramide levels, a response not observed in skeletal muscle tissue. Plasma ceramides (e.g., C24:0-ceramide) were positively correlated with plasma NEFA and milk yield, and positively correlated with NEFA levels following a glucose challenge. Our data demonstrate a remodeled plasma and hepatic sphingolipidome in mid-lactation dairy cows fed PALM. The potential involvement in ceramide in homeorhetic nutrient partitioning to support lactation requires further consideration.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Excessive Autophagy Activation and Increased Apoptosis Are Associated with -Induced Cardiomyocyte .

Diabetic cardiomyopathy (DCM) remains the major cause of death associated with diabetes. Researchers have demonstrated the importance of impaired cardiac signaling in this process. (IR) is an important predictor of DCM. Previous studies examining the dynamic changes in autophagy during IR have yielded inconsistent results. This study aimed to investigate the dynamic changes in autophagy and apoptosis in the rat H9c2 cardiomyocyte IR model. H9c2 cells were treated with 500\u2009M (PA) for 24 hours, resulting in the induction of IR. To examine autophagy, monodansylcadaverine staining, GFP-LC3 puncta confocal observation, and Western blot analysis of LC3I-to-LC3II conversion were used. Results of these studies showed that autophagic vesicles increased in numbers during the first 24 hours and then decreased by 36 hours after PA treatment. Western blot analysis showed that treatment of H9c2 cells with 500\u2009M PA for 24 hours decreased the expression of Atg12-Atg5, Atg16L1, Atg3, and PI3Kp85. Annexin V/PI flow cytometry revealed that PA exposure for 24 hours increased the rate of apoptosis. Together, this study demonstrates that PA induces IR in H9c2 cells and that this process is accompanied by excessive activation of autophagy and increases in apoptosis.

Keyword: insulin resistance

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver-body weight ratio, blood biochemistry, and in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: insulin resistance

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Metabolism Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, , and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic metabolism. DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: insulin resistance

Ameliorative effect of panaxynol on the reduction in high-molecular-weight adiponectin secretion from 3T3-L1 adipocytes treated with acids.

Reduced plasma levels of the high-molecular weight (HMW) form of adiponectin, rather than total adiponectin levels, have been shown to be closely associated with various metabolic diseases including , type 2 diabetes, and cardiovascular disease. Therefore, we sought to explore active, naturally occurring compounds that promote the recovery of HMW adiponectin secretion suppressed by in our model. A total of 90 crude drug extracts were screened for the ability to augment HMW adiponectin secretion from 3T3-L1 adipocytes treated with . Panaxynol was isolated from Saposhnikovia divaricata as an active compound with HMW adiponectin promoting properties. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists are reported to increase the secretion of HMW adiponectin, although the effects of panaxynol were found to be independent of PPARγ activation. When the underlying mechanisms were further examined, panaxynol was found to inhibit the -induced downregulation of forkhead box O1 (FoxO1) protein, and the anti-lipotoxic effects were abolished by a FoxO1 inhibitor. Furthermore, CCAAT/enhancer-binding protein-α (C/EBPα) mRNA levels were also increased by panaxynol. Reactive oxygen species have critical roles in the reduction in HMW adiponection secretion by ; however, panaxynol reduced this increase in reactive oxygen species generation, followed by reductions in markers of endoplasmic reticulum stress and inflammation. Taken together, these findings suggest that panaxynol ameliorates the impaired HMW adiponection secretion in adipocytes treated with by restoring FoxO1 expression, owing to inhibition of reactive oxygen species generation, in a PPARγ-independent manner.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Ganoderma Lucidum Polysaccharide Peptide Alleviates Hepatoteatosis via Modulating Bile Metabolism Dependent on FXR-SHP/FGF.

Non-alcoholic fatty liver disease (NAFLD) encompasses a series of pathologic changes ranging from steatosis to steatohepatitis, which may progress to cirrhosis and hepatocellular carcinoma. The purpose of this study was to determine whether ganoderma lucidum polysaccharide peptide (GLPP) has therapeutic effect on NAFLD.Ob/ ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic pathways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blot. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD.GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, liver dysfunction and liver . Pathways of glycerophospholipid metabolism, fatty metabolism and primary bile biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1, CYP8B1, FXR, SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited fatty synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepatocytes induced by oleic and .GLPP significantly improves NAFLD via regulating bile synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits fatty synthesis, indicating that GLPP might be developed as a therapeutic drug for NAFLD.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: insulin resistance

Theaflavins Improve Sensitivity through Regulating Mitochondrial Biosynthesis in -Induced HepG2 Cells.

Theaflavins, the characteristic and bioactive polyphenols in black tea, possess the potential improving effects on -associated metabolic abnormalities, including obesity and type 2 diabetes mellitus. However, the related molecular mechanisms are still unclear. In this research, we investigated the protective effects of theaflavins against in HepG2 cells induced by . Theaflavins significantly increased glucose uptake of -resistant cells at noncytotoxic doses. This activity was mediated by upregulating the total and membrane bound glucose transporter 4 protein expressions, increasing the phosphor-Akt (Ser473) level, and decreasing the phosphorylation of IRS-1 at Ser307. Moreover, theaflavins were found to enhance the mitochondrial DNA copy number, down-regulate the PGC-1β mRNA level and increase the PRC mRNA expression. Mdivi-1, a selective mitochondrial division inhibitor, could attenuate TFs-induced promotion of glucose uptake in -resistant HepG2 cells. Taken together, these results suggested that theaflavins could improve hepatocellular induced by free fatty acids, at least partly through promoting mitochondrial biogenesis. Theaflavins are promising functional food ingredients and medicines for improving -related disorders.

Keyword: insulin resistance

Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and -sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether obesity impacts placental lipid transport and metabolism remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and -sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal obesity. Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in lipid classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to and low-grade inflammation in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by obesity.-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and -sensitizing properties.

Keyword: insulin resistance

[How surplus of fatty in food initiates hypertriglyceridemia, increases cholesterol of low density lipoproteins, triggers atherosclerosis and develops atheromatosis.]

In phylogenesis, the first transfer of all fatty acids to cells is implemented by high density lipoproteins. Later, unsaturated and polyene fatty acids are transferred to cell by low density lipoproteins. The -depended cells absorb saturated fatty , oleic mono-unsaturated fatty and of the same name triglycerides in very low density lipoproteins. The hepatocytes secrete , oleic and linoleic very low density lipoproteins separately. In blood, under hydrolysis of triglycerides, cells absorb ligand and oleic very low density lipoproteins by force of апоЕ/В-100 endocytosis; they are not transformed into low density lipoproteins. The saturated fatty acids in the form of polyether of cholesterol turn into linoleic very low density lipoproteins from high density lipoproteins at impact of protein transferring polyene ethers of cholesterol. They transform very low density lipoproteins into low density lipoproteins of the same name; the cells absorb them by force of апоЕ/В-100 endocytosis. In physiological sense, amount of oleic very low density lipoproteins are always more than of very low density lipoproteins. Under syndrome of there is no transformation of saturated fatty synthesized from glucose in vivo into oleic mono-saturated fatty . The hepatocytes secrete into blood mainly very low density lipoproteins which amount exceeds oleic very low density lipoproteins. Under slow hydrolysis in blood, main mass of very low density lipoproteins becomes low density lipoproteins. These very lipoproteins initiate hyperlipidemia, increase content of cholesterol of cholesterol-low density lipoproteins, lower cholesterol-high density lipoproteins, decrease bio-availability of polyene fatty acids for cells, trigger development of atherosclerosis and formation of atheromatosis in intima of arteries. The aphysiologic effect of surplus of saturated fatty in vivo and triglycerides of the same name can\'t be eliminated under increasing of content of ω-3 polyene fatty acids in food and effect of statines. All this is to be rationally applied in prevention of hypertriglyceridemia, atherosclerosis, atheromatosis of coronary arteries, ischemic heart disease and myocardium infarction.

Keyword: insulin resistance

Individual free fatty acids have unique associations with inflammatory biomarkers, and secretion in healthy and gestational diabetic pregnant women.

We investigated the relationships of maternal circulating individual free fatty acids (FFA) with , secretion and inflammatory biomarkers during mid-pregnancy.The data were drawn from a prospective cohort of generally healthy pregnant women (n=1368, African-American 36%, Hispanic 48%, Caucasian 16%) in Camden, NJ. We quantitatively determined 11 FFAs, seven cytokine/adipokine, homeostatic model assessment of (HOMA-IR) and C-peptide levels from the fasting blood samples that were collected at 16 weeks of gestation. Multivariate analyses were performed along with separate analyses for each individual FFA.High HOMA-IR (p<0.001) and C-peptide (p<0.0001) levels were positively associated with a twofold to fourfold increased risk for developing gestational diabetes mellitus (GDM). Negative relationships were found with specific FFAs (molecular percentage, palmitoleic, oleic, linolenic, myristic acids) and HOMA-IR and C-peptide levels (p<0.01\u2009to p<0.0001). In contrast, , stearic, arachidonic, dihomo-γ-linolenic (DGLA) and docosahexaenoic acids were positively associated with HOMA-IR and C-peptide (p<0.01\u2009to p<0.0001). The individual FFAs also predicted cytokine/adipokine levels. For example, women who had elevated DGLA (highest quartile) were twice as (adjusted OR 2.06, 95%\u2009CI 1.42 to 2.98) likely to have higher interleukin (IL)-8 (p<0.0001) levels. Conversely, women with high palmitoleic, oleic, and linolenic levels had reduced odds (≥2-fold, p<0.01\u2009to p<0.001) for having higher IL-8, IL-6 or tumor necrosis factor-alpha levels.Our results suggest that maternal individual FFAs uniquely affect and secretion. The effects are either direct or indirect via modulation of the inflammatory response. Modifying the composition of FFAs may help in reducing the risk of GDM.

Keyword: insulin resistance

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid metabolism-related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and tolerance tests, and immunohistochemistry. The expression of lipid metabolism- and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on release in co-cultures with isolated rat islets.HFD increased body weight and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and , upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid metabolism-related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid metabolism, suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

A high-fat diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high-fat diet (HFD) commonly leads to obesity and . However, even 3\u2009days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus.Mice were fed either a low-fat diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from fat for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose metabolism and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: insulin resistance

MicroRNA‑29a is involved lipid metabolism dysfunction and in C2C12 myotubes by targeting PPARδ.

MicroRNA‑29a (miR‑29a) expression has been reported to be closely associated with skeletal muscle and type 2 diabetes. The present study investigated the effect of miR‑29a on (PA)‑induced lipid metabolism dysfunction and in C2C12 myotubes via overexpressing or silencing of miR‑29a expression. Mouse C2C12 myoblasts were cultured, differentiated and transfected with miR‑29a or miR‑29a inhibitor lentiviral with or without subsequent (PA) treatment. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were performed to assess the mRNA and protein levels of related genes, respectively. PA treatment increased the expression of miR‑29a in a time‑ and dose‑ dependent manner. miR‑29a silencing improved ‑induced glucose uptake and increased glucose transporter‑4 (GLUT4) transportation to the plasma membrane by upregulating its target peroxisome proliferator‑activated receptor δ (PPARδ). Furthermore, it was observed that miR‑29a regulated the expression of genes associated with lipid metabolism, including pyruvate dehydrogenase kinase isoform, mitochondrial uncoupling protein (UCP)2, UCP3, long chain specific acyl‑CoA dehydrogenase, mitochondrial and fatty transport protein 2. The results confirmed that silencing miR‑29a induced a decrease in glucose transport and affected lipid metabolism in PA‑treated C2C12 cells, and therefore may be involved in by targeting PPARδ in skeletal muscle. Therefore, the inhibition of miR‑29a may be a potential novel strategy for treating and type 2 diabetes.

Keyword: insulin resistance

[The physical chemical and biological features of triglycerides. The cell absorption of functionally different +oleic lipoproteins of very low and density and linoleic+linolenic lipoproteins of low density.]

The earlier -independent low-density lipoproteins and more late -dependent very low-density lipoproteins implement different functions at the stages of phylogenesis. The disorder of biological function of trophology, alteration of fatty acids in triglycerides, prevalence of very low-density lipoproteins over oleic very low-density lipoproteins supply mitochondria of cells with non-optimal substrate - saturated fatty for gaining energy, ATP synthesis. Physiologically, cells implement oleic alternative of fatty acids metabolism, oxidizing mainly ω-9 endogenous oleic mono-unsaturated fatty . The pathology of low density lipoproteins is primary deficiency of poly-unsaturated fatty acids in cells, atherosclerosis and atheromotosis of intima of arteries of elastic type with development of dense plaques from poly-unsaturated fatty acids in the form of polyethers of cholesterol. The pathology of very low-density lipoproteins includes: a) syndrome of to ; b) pathology of phylogenetically earlier -independent visceral fatty tissue - metabolic syndrome; c) pathology of phylogenetically later -dependent subcutaneous adipocytes - obesity; d) secondary atherosclerosis, under cumulation of low-density lipoproteins in blood with development of atherothrombosis of intima of arteries, soft plaques rich with triglycerides. As for the prevention of disorders of transfer of fatty acids to very low-density lipoproteins and low-density lipoproteins is common in many ways - minimization of aphysiological effect of surplus amount of food, biological function of diet. The prevention at the level of population includes: a) maximal limitation of content of saturated fatty in food; b) moderate increasing of polysaturated fatty acids, ω-3 poly-saturated fatty acids predominantly; c) increasing of physical activity. The pharmaceuticals are not provided by biology in primary prevention of metabolic pandemics under aphysiological impact of environment factors.

Keyword: insulin resistance

Fish oil supplementation inhibits endoplasmic reticulum stress and improves : involvement of AMP-activated protein kinase.

The beneficial effects of fish oil consumption on glucose metabolism have been generally reported. However, the mechanism underlying the fish oil-induced protective effects against remains unclear. Endoplasmic reticulum (ER) stress is recognized as an important contributor to . The aim of this study is to evaluate whether fish oil supplementation reduces ER stress and ameliorates in diet-induced obese mice, and to investigate the molecular mechanism of fish oil-induced benefits on ER stress. C57BL/6J mice were fed one of the following diets for 12 weeks: the low-fat diet (LFD), the high-fat diet (HFD) or the fish oil-supplemented high-fat diet (FOD). Fish oil supplementation led to lower blood glucose, better glucose tolerance and improved sensitivity in high-fat diet-induced obese mice. Importantly, fish oil administration inhibited high-fat feeding-induced ER stress and reduced adipose tissue dysfunction. The fish oil-induced improvements were accompanied by the elevation of phosphorylated AMP-activated protein kinase (AMPK) expression in white adipose tissue. Correspondingly, the results of in vitro experiments showed that docosahexaenoic (DHA), the main n-3 polyunsaturated fatty (PUFA) in the fish oil used in the study, led to a dose-dependent increase in AMPK phosphorylation and suppressed (PA)-triggered ER stress in differentiated 3T3-L1 adipocytes. Furthermore, AMPK inhibitor (compound C) treatment largely blocked the effects of DHA to inhibit PA-induced ER stress. Our data indicate that n-3 PUFAs suppress ER stress in adipocytes through AMPK activation, and may thereby exert protective effects against high-fat feeding-induced adipose tissue dysfunction and .

Keyword: insulin resistance

[Laurine fatty acids, medium fatty acids and triglycerides, , resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: insulin resistance

Saturated fatty acids induce development of both metabolic syndrome and osteoarthritis in rats.

The predominant saturated fatty acids (SFA) in human diets are lauric (LA, C12:0), myristic (MA, C14:0), (PA, C16:0) and stearic (SA, C18:0). The aim of this study was to investigate whether diets containing individual SFA together with excess simple carbohydrates induce osteoarthritis (OA)-like changes in knee joints and signs of metabolic syndrome in rats. Rats were given either a corn starch diet or a diet composed of simple carbohydrates together with 20% LA, MA, PA, SA or beef tallow for 16 weeks. Rats fed beef tallow, SA, MA or PA diets developed signs of metabolic syndrome, and also exhibited cartilage degradation and subchondral bone changes similar to OA. In contrast, replacement of beef tallow with LA decreased signs of metabolic syndrome together with decreased cartilage degradation. Furthermore, PA and SA but not LA increased release of matrix sulphated proteoglycans in cultures of bovine cartilage explants or human chondrocytes. In conclusion, we have shown that longer-chain dietary SFA in rats induce both metabolic syndrome and OA-like knee changes. Thus, diets containing SFA are strongly relevant to the development or prevention of both OA and metabolic syndrome.

Keyword: insulin resistance

[The cognitive biological function, difference in etiology and generality of pathogenesis of metabolic pandemics. The metabolism regulation in carnivorous and herbivorous animals under effect of environment factors.]

In phylogenesis, carnivorous animals initially and later herbivorous animals formed separate transfer of exogenous fatty acids in blood, predominantly and later - endogenous oleic pool, synthesized in hepatocytes from glucose. In vivo, exogenous fatty acids transfer α-lipoproteins to all cells and chylomicrons to hepatocytes. The endogenous oleic fatty , synthesized in hepatocytes is transferred to all cells by β-lipoproteins and to -dependent lipoproteins of very low density. The limitation of cognitive biological function in herbivorous animals underlies the process of transferring of surplus amount of exogenous and alcohol cholesterol by β-lipoproteins becoming a non-physiological one. The bio-availability is affected concerning cells of exogenous polyene fatty acids that transfer in blood α-lipoproteins in carnivorous animals and at the first stage in herbivorous animals. The formation of non-ligand very low density lipoproteins-β-proteins is affected and their collection and partial utilization in intima generates lipoidosis. In summary, all this becomes a cause of formation of atherosclerosis and atheromatosis. The limitations in phylogenesis of cognitive biological function are considered as an etiological factor of metabolic pandemics. The etiological factors are specific for every one of metabolic pandemics and pathogenesis of all pandemics is developed following common algorithm. It is likely that metabolic pandemics (atherosclerosis and atheromatosis, metabolic essential arterial hypertension, metabolic syndrome, syndrome of , obesity and non-alcoholic fatty disease of liver)are quite not identical to nosologic forms of diseases if considered according etiological factors, pathogenesis and pathology rate in population. The exclusion of all inherent forms of disorder of metabolism of fatty acids and their transfer in β-lipoproteins demonstrates that problem of atherosclerosis, atheromatosis, coronarosclerosis and ischemic heart disease mainly turns out to be a case of eaten one\'s fill , a result of environment factors impact, disorder of biological function of trophology, biological reaction of exotrophy (external nutrition) and biological function of homeostasis. The prevention of atherosclerosis, atheromatosis and ischemic heart disease in population will be effective only under condition of obligatory individual support of actioning cognitive biological function by more complete biological function of intelligence. The active prevention of metabolic pandemics by force of personal development of cognitive biological function, intelligence function, provides no application of pharmaceuticals. The disorder of cognitive biological function is needed to be improved, incorrect positioning of Homo Sapiens species as omnivorous in animal world; Homo Sapiens is a herbivorous one.

Keyword: insulin resistance

Resveratrol reduces liver endoplasmic reticulum stress and improves sensitivity in vivo and in vitro.

: The aim of the study was to examine the effects of resveratrol upon hepatic endoplasmic reticulum stress (ERS) and sensitivity in vivo and in vitro. : C57BL/6J mice were fed a high-fat diet (HFD) for 8 weeks, and was evaluated by the intraperitoneal glucose tolerance test (IPGTT). Mice were then treated with resveratrol for 12 weeks and blood and liver samples collected. Blood biochemical indicators were determined by kits, liver protein expression was determined by western blot, and morphological changes were observed by histological staining. (PA)-induced -resistant HepG2 cells were established. Cells were exposed to 100, 50 or 20 μM resveratrol for 24 hrs, and proliferation/cytotoxicity was determined. Cells were divided into five groups: control, PA, PA + Rev (100 μM), PA + Rev (50 μM) and PA + Rev (20 μM) groups. After 24 hrs of treatment, cellular proteins were analyzed the same way as animal tissues. : The IPGTT confirmed that the model was established successfully. After resveratrol treatment, fasting blood glucose and cholesterol levels declined and the quantitative sensitivity check index increased. Western-blot results showed that resveratrol-treated HFD mice had reduced hepatic levels of p-PERK, ATF-4 and TRIB3, and increased the levels of ATF-6, p-AKT and p-GSK3β. In the cell model, resveratrol with 100 and 50 μM enhanced ERS and , whereas 20 μM had beneficial effects, similar to the animal model. : Resveratrol reduced hepatic ERS, thereby improving sensitivity and glucose levels. However, high doses of resveratrol had harmful effects on cells, elevating ERS and . The safe dose of resveratrol needs further investigation.

Keyword: insulin resistance

Inhibitory effect of 17β‑estradiol on triglyceride synthesis in skeletal muscle cells is dependent on ESR1 and not ESR2.

The present study aimed to investigate the inhibitory effects and the mechanisms underlying 17β‑estradiol (E2) effects on triglyceride synthesis and in skeletal muscle tissues and cells. Ovariectomy (OVX) was performed on 6‑month‑old female rats treated with or without E2. Subsequently, various serum biochemical markers were measured. Additionally, pathological alterations of the uterus, liver and skeletal muscle were analyzed, and the content of triglycerides (TG) in muscle was detected. Differentiated myotubes formed by C2C12 cells were treated with (PA) or pretreated with E2, estrogen receptor (ESR)\xa01 agonist propylpyrazoletriol (PPT) and ESR2 agonist diarylpropionitrile (DPN). Subsequently, the mRNA or protein expression levels of ESR1/2, peroxisome proliferator activated receptor α (PPARα), CD36 molecule (CD36), fatty synthase (FASN), perilipin 2 (PLIN2), phosphorylated acetyl‑CoA carboxylase α (p‑ACACA), p‑AKT serine/threonine kinase (p‑AKT) and p‑mitogen‑activated protein kinase\xa08 (p‑MAPK8) were analyzed in skeletal muscle or in C2C12 cells by reverse transcription‑semi‑quantitative polymerase chain reaction and western blotting. The present results suggested that treatment with E2 inhibited OVX‑induced body weight gain, TG accumulation and . The protein or mRNA expression levels of ESR1, CD36, PPARα, p‑ACACA and p‑AKT were decreased, whereas the protein or mRNA expression levels of ESR2, PLIN2, FASN and p‑MAPK8 were increased in the OVX group. Of note, treatment with E2 restored the expression levels of the aforementioned factors. In C2C12 cells, treatment with E2 or PPT reversed the alterations induced by treatment with PA. In contrast, pretreatment with DPN did not influence the effect of PA. Collectively, E2 was able to interact with ESR1, thus activating the CD36‑PPARα pathway, decreasing the level of TG in the muscles and improving in skeletal muscles and C2C12 cells.

Keyword: insulin resistance

Induction of Sphk1 activity in obese adipose tissue macrophages promotes survival.

During obesity, adipose tissue macrophages (ATM) are increased in concert with local inflammation and . Since the levels of sphingolipid (SLs) in adipose tissue (AT) are altered during obesity we investigated the potential impact of SLs on ATMs. For this, we first analyzed expression of SL metabolizing genes in ATMs isolated from obese mice. A marked induction of sphingosine kinase 1 (Sphk1) expression was observed in obese ATM when compared to lean ATM. This induction was observed in both MGL-ve (M1) and MGL1+ve (M2) macrophages from obese WAT. Next, RAW264.7 cells were exposed to excessive palmitate, resulting in a similar induction of Sphk1. This Sphk1 induction was also observed when cells were treated with chloroquine, a lysosomotropic amine impacting lysosome function. Simultaneous incubation of RAW cells with palmitate and the Sphk1 inhibitor SK1-I promoted cell death, suggesting a protective role of Sphk1 during lipotoxic conditions. Interestingly, a reduction of endoplasmic reticulum (ER) stress related genes was detected in obese ATM and was found to be associated with elevated Sphk1 expression. Altogether, our data suggest that lipid overload in ATM induces Sphk1, which promotes cell viability.

Keyword: insulin resistance

Reduces the Autophagic Flux and Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high fat diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related metabolic diseases such as type II diabetes mellitus (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: insulin resistance

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding . These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: insulin resistance

METRNL attenuates lipid-induced inflammation and via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, , and diabetes. Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and . Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: insulin resistance

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the liver of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in liver cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the liver steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and liver lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: insulin resistance

Fatty acids, epigenetic mechanisms and chronic diseases: a systematic review.

Chronic illnesses like obesity, type 2 diabetes (T2D) and cardiovascular diseases, are worldwide major causes of morbidity and mortality. These pathological conditions involve interactions between environmental, genetic, and epigenetic factors. Recent advances in nutriepigenomics are contributing to clarify the role of some nutritional factors, including dietary fatty acids in gene expression regulation. This systematic review assesses currently available information concerning the role of the different fatty acids on epigenetic mechanisms that affect the development of chronic diseases or induce protective effects on metabolic alterations.A targeted search was conducted in the PubMed/Medline databases using the keywords "fatty acids and epigenetic". The data were analyzed according to the PRISMA-P guidelines.Consumption fatty acids like n-3 PUFA: EPA and DHA, and MUFA: oleic and palmitoleic was associated with an improvement of metabolic alterations. On the other hand, fatty acids that have been associated with the presence or development of obesity, T2D, pro-inflammatory profile, atherosclerosis and IR were n-6 PUFA, saturated fatty acids (stearic and ), and trans fatty acids (elaidic), have been also linked with epigenetic changes.Fatty acids can regulate gene expression by modifying epigenetic mechanisms and consequently result in positive or negative impacts on metabolic outcomes.

Keyword: insulin resistance

Anxa2 gene silencing attenuates obesity-induced by suppressing the NF-κB signaling pathway.

(IR) continues to pose a major threat to public health due to its role in the pathogenesis of metabolic syndrome and its ever-increasing prevalence on a global scale. The aim of the current study was to investigate the efficacy of Anxa2 in obesity-induced IR through the mediation of the NF-κB signaling pathway. Microarray analysis was performed to screen differentially expressed genes associated with obesity. To verify whether Anxa2 was differentially expressed in IR triggered by obesity, IR mouse models were established in connection with a high-fat diet (HFD). In the mouse IR model, the role of differentially expressed Anxa2 in glycometabolism and IR was subsequently detected. To investigate the effect of Anxa2 on IR and its correlation with inflammation, a (PA)-induced IR cell model was established, with the relationship between Anxa2 and the NF-κB signaling pathway investigated accordingly. Anxa2 was determined to be highly expressed in IR. Silencing Anxa2 was shown to inhibit IR triggered by obesity. When Anxa2 was knocked down, elevated expression of phosphorylated receptor substrate 1 (IRS1), IRS1 and peroxisome proliferator-activated receptor coactivator-1a, and glucose tolerance and sensitivity along with 2-deoxy-d-glucose uptake was detected, whereas decreased expression of suppressor of cytokine signaling 3, IL-6, IL-1β, TNF-α, and p50 was observed. Taken together, the current study ultimately demonstrated that Anxa2 may be a novel drug strategy for IR disruption, indicating that Anxa2 gene silencing is capable of alleviating PA or HFD-induced IR and inflammation through its negative regulatory role in the process of p50 nuclear translocation of the NF-κB signaling pathway.

Keyword: insulin resistance

Adipocyte-Derived Exosomal MiR-27a Induces in Skeletal Muscle Through Repression of PPARγ.

The mechanism by which adipocyte-derived endocrine factors promote in skeletal muscle are not fully understood. MiR-27a is highly expressed in sera of obese individuals with prediabetes and T2DM, and mainly derived by adipose tissues. Thus, miR-27a secreted into circulation by adipose tissue may regulate in skeletal muscle. The association between miR-27a and in skeletal muscle was determined in obese children, high-fat diet-induced miR-27a knockdown obese mice, db/db mice and C2C12 cells overexpressing miR-27a. The crosstalk mediated by exosomal miR-27a between adipose tissue and skeletal muscle was determined in C2C12 cells incubated with conditioned medium prepared from palmitate-treated 3T3-L1 adipocytes. We showed that serum miR-27a level correlated positively with obesity and in obese children, and that elevated serum miR-27a levels correlated with in leptin receptor-deficient db/db mice, and with obesity and in high-fat diet-fed C57BL/6J mice. MiR-27a released from adipocytes of high-fat diet-fed C57BL/6J mice was associated with triglyceride accumulation. MiR-27a derived from these adipocytes induced in C2C12 skeletal muscle cells through miR-27a-mediated repression of PPARγ and its downstream genes involved in the development of obesity. These results identify a novel crosstalk signaling pathway between adipose tissue and skeletal muscle in the development of , and indicate that adipose tissue-derived miR-27a may play a key role in the development of obesity-triggered in skeletal muscle.

Keyword: insulin resistance

Upregulated absorption of dietary acids with changes in intestinal transporters in non-alcoholic steatohepatitis (NASH).

is an important risk factor for the pathogenesis of non-alcoholic steatohepatitis (NASH), but changes in intestinal absorption in NASH are unclear. The aim of this study was to clarify changes in intestinal absorption and their association with the pathogenesis of NASH.A total of 106 participants were recruited to the study, of whom 33 were control subjects (control group), 32 were patients with NASH Brunt stage 1-2 [early NASH (e-NASH)], and 41 were patients with NASH Brunt stage 3-4 [advanced NASH (a-NASH)]. C-labeled palmitate was administered directly into the duodenum of all participants by gastrointestinal endoscopy. Breath CO levels were measured to quantify absorption, and serum Apolipoprotein B-48 (ApoB-48) concentrations were measured after a test meal to quantify absorbed chylomicrons. Expressions of fatty (FA) transporters were also examined. The associations of breath CO levels with hepatic steatosis, fibrosis and was evaluated using laboratory data, elastography results and liver histology findings.Overall, CO excretion was significantly higher in e-NASH patients than in the control subjects and a-NASH patients (P\xa0<\xa00.01). e-NASH patients had higher serum ApoB-48 levels, indicating increased transport via chylomicrons in these patients. Jejunal mRNA and protein expressions of microsomal triglyceride transfer protein and cluster of differentiation 36 were also increased in both NASH patient groups. The CO excretion of e-NASH patients was significantly correlated with the degree of hepatic steatosis, fibrosis and (P\xa0=\xa00.005, P\xa0<\xa00.001, P\xa0=\xa00.019, respectively).Significantly upregulated absorption by activation of its transporters was evident in patients with NASH, and clinical progression of NASH was related to absorption. These dietary changes are associated with the onset and progression of NASH.

Keyword: insulin resistance

Celastrol Reverses -Induced in HepG2 Cells via Restoring the miR-223 and GLUT4 Pathway.

The natural triterpenoid compound celastrol ameliorates (IR) in animal models, but the underlying molecular mechanism is unclear. In this study, we investigated how celastrol regulates IR.The HepG2 cellular IR model was initially established with (PA). The expression and activity of glucose transporter 4 (GLUT4), receptor substrate-1 (IRS1) and 9 microRNAs (miRNAs) (miR-7, -34a, -96, -113, -126, -145, -150, -223 and -370) were detected before and after celastrol treatment using the PA-induced HepG2 IR model.The results showed that 250\u2009µM PA for ≥2 days was optimal for inducing IR in HepG2 cells; 600\u2009nM celastrol significantly attenuated the PA-induced IR in HepG2 cells. The PA-induced GLUT4 and IRS1 downregulation and Ser307 phosphorylation on IRS1 was reversed by subsequent treatment with 600\u2009nM celastrol for 6\u2009h. We next investigated which IR-related miRNAs were possible upstream regulators of celastrol-mediated reversal of PA-induced HepG2 IR. Two miRNAs, miR-150 and -223, were significantly downregulated by PA and were re-raised by subsequent celastrol treatment; and miR-223 was upstream of miR-150. Moreover, knocking down miR-223 abolished celastrol\'s anti-IR effects in the PA-induced model.Collectively, our results demonstrated that celastrol reverses PA-induced IR-related alterations, in part via miR-223 in HepG2 cells. Further investigation is warranted for establishing the clinical potential of celastrol in treating IR-related disorders.Copyright © 2019. Published by Elsevier Inc.

Keyword: insulin resistance

STING-IRF3 Triggers Endothelial Inflammation in Response to Free Fatty -Induced Mitochondrial Damage in Diet-Induced Obesity.

Metabolic stress in obesity induces endothelial inflammation and activation, which initiates adipose tissue inflammation, , and cardiovascular diseases. However, the mechanisms underlying endothelial inflammation induction are not completely understood. Stimulator of interferon genes (STING) is an important molecule in immunity and inflammation. In the present study, we sought to determine the role of STING in -induced endothelial activation/inflammation.In cultured endothelial cells, treatment activated STING, as indicated by its perinuclear translocation and binding to interferon regulatory factor 3 (IRF3), leading to IRF3 phosphorylation and nuclear translocation. The activated IRF3 bound to the promoter of ICAM-1 (intercellular adhesion molecule 1) and induced ICAM-1 expression and monocyte-endothelial cell adhesion. When analyzing the upstream signaling, we found that activated STING by inducing mitochondrial damage. treatment caused mitochondrial damage and leakage of mitochondrial DNA into the cytosol. Through the cytosolic DNA sensor cGAS (cyclic GMP-AMP synthase), the mitochondrial damage and leaked cytosolic mitochondrial DNA activated the STING-IRF3 pathway and increased ICAM-1 expression. In mice with diet-induced obesity, the STING-IRF3 pathway was activated in adipose tissue. However, STING deficiency ( ) partially prevented diet-induced adipose tissue inflammation, obesity, , and glucose intolerance.The mitochondrial damage-cGAS-STING-IRF3 pathway is critically involved in metabolic stress-induced endothelial inflammation. STING may be a potential therapeutic target for preventing cardiovascular diseases and in obese individuals.© 2017 American Heart Association, Inc.

Keyword: insulin resistance

TAp63 is correlated with chronic inflammation in patients with newly diagnosed type 2 diabetes mellitus.

To investigate TAp63 expression in patients with type 2 diabetes mellitus (T2DM) and the potential correlations between TAp63 and proinflammatory cytokines production and other clinical parameters.Peripheral blood mononuclear cells (PBMCs) and plasma were collected from 72 T2DM (cases) and 72 healthy subjects (controls). Fasting blood glucose (FBG), fasting (FIN) and a blood lipid profile were measured. The homeostasis model assessment (HOMA) was used to estimate (IR). Plasma tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were determined. PBMCs isolated from healthy subjects were cultured with or without 33.3 mmol/l glucose or 0.5 mmol/l (PA) for 6 h, 24 h, 48 h, and 72 h. The expression of TAp63 at mRNA and protein levels in PBMCs was analyzed using real-time qRT-PCR and western blots, respectively.TAp63 expression was significantly lower in T2DM patients compared with that of the controls. In addition, TAp63 expression showed a negative correlation with FBG, FIN, HbA1c, HOMA-IR, FFAs, TNF-α, and IL-6 levels. Treatment with 33.3 mmol/l glucose or 0.5 mmol/l PA increased TAp63 expression in the cultured PBMCs.TAp63 level may be correlated with chronic inflammatory state and perturbed glucose and lipid metabolism in T2DM.Copyright © 2018. Published by Elsevier Inc.

Keyword: insulin resistance

Hydroxystearic Acids Activate GPR40, Which Is Involved in Their Beneficial Effects on Glucose Homeostasis.

hydroxystearic acids (PAHSAs) are endogenous lipids with anti-diabetic and anti-inflammatory effects. PAHSA levels are reduced in serum and adipose tissue of -resistant people and high-fat diet (HFD)-fed mice. Here, we investigated whether chronic PAHSA treatment enhances sensitivity and which receptors mediate PAHSA effects. Chronic PAHSA administration in chow- and HFD-fed mice raises serum and tissue PAHSA levels ∼1.4- to 3-fold. This improves sensitivity and glucose tolerance without altering body weight. PAHSA administration in chow-fed, but not HFD-fed, mice augments and glucagon-like peptide (GLP-1) secretion. PAHSAs are selective agonists for GPR40, increasing Ca flux, but not intracellular cyclic AMP. Blocking GPR40 reverses improvements in glucose tolerance and sensitivity in PAHSA-treated chow- and HFD-fed mice and directly inhibits PAHSA augmentation of glucose-stimulated secretion in human islets. In contrast, GLP-1 receptor blockade in PAHSA-treated chow-fed mice reduces PAHSA effects on glucose tolerance, but not on sensitivity. Thus, PAHSAs activate GPR40, which is involved in their beneficial metabolic effects.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Resveratrol affects hepatic gluconeogenesis via histone deacetylase 4.

The aim of this study was to determine whether resveratrol (Rev) affects the expression, phosphorylation, and nuclear and cytoplasmic distribution of histone deacetylase 4 (HDAC4), which in turn affects gluconeogenesis in hepatocytes under an -resistant state.HepG2 cells were treated with 0.25 mmol/L (PA) to establish an model. The cells were divided into five groups: control, PA, PA + Rev 100 µM, PA + Rev 50 µM, and PA + Rev 20 µM. After treatment for 24 hours, mRNA and protein expression levels of gluconeogenesis pathway-related molecules and HDAC4 were examined. Next, HepG2 cells were transfected with siRNA-HDAC4. The cells were divided into control, PA, PA + Rev 20 µM, PA + Rev 20 µM +siRNA-HDAC4 negative control, and PA + Rev 20 µM +siRNA-HDAC4 knockdown groups to determine the expression of gluconeogenesis pathway proteins.Compared with the control group, the gluconeogenesis pathway-related molecules, glucose-6-phosphatase catalytic subunit (G6PC), phosphoenolpyruvate carboxykinase 1 (PCK1) and forkhead box protein O1 (FOXO1), were increased, and the phosphorylation of FOXO1 decreased after PA treatment. The p-HDAC4 level decreased with the increase in HDAC4 in the nucleus and the decrease in HDAC4 in the cytoplasm in the PA group. Treatment with Rev 20 µM suppressed gluconeogenesis and promoted HDAC4 shuttling into the cytoplasm from the nucleus. However, 100 and Rev 50 µM exerted the opposite effects. Finally, after HDAC4 knockdown, the expression levels of the key gluconeogenesis molecules, G6PC, PCK1, and FOXO1, were increased, and p-FOXO1 was decreased, indicating that gluconeogenesis was enhanced.A low concentration of Rev inhibited gluconeogenesis under conditions via translocation of HDAC4 from the nucleus to the cytoplasm.

Keyword: insulin resistance

Pioglitazone Enhances Cytosolic Lipolysis, β-oxidation and Autophagy to Ameliorate Hepatic Steatosis.

Non-alcoholic fatty liver disease closely contributes to the development of obesity and . Even though pioglitazone has been reported to effectively lessen hepatic steatosis in human studies, its molecular mechanism remains unclear. This study is designed to investigate the regulation of cytosolic lipolysis, β-oxidation and autophagy by pioglitazone in a mice model of high fat diet (HFD) and cell model incubated with . Our results revealed hepatic steatosis was apparently induced by HFD and it was significantly reversed by pioglitazone. The serum and hepatic triglyceride content was significantly decreased by co-administered pioglitazone with HFD. Hepatic expression of cytosolic-lipolysis related proteins (ATGL, HSL), β-oxidation (CPT-1A) and autophagy-related proteins (ATG7, LC3, LAL) was significantly enhanced by pioglitazone. Knockdown PPARα/PPARγ in AML12 cells significantly and proportionally reduced the expressions of ATGL, CPT-1A and LC3II, which was induced by pioglitazone. Furthermore, facilitation of the autophagic flux by pioglitazone was obviously blocked by lysosomal inhibitor, leupeptin, to demonstrate accumulation of the LC3II and intracellular lipid in AML12 cells. Our results demonstrated that pioglitazone attenuating the hepatic steatosis may be mediated by enhancing cytosolic lipolysis, β-oxidation and autophagy in a PPARα and PPARγ dependent manner.

Keyword: insulin resistance

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic and Type 2 diabetes (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo lipogenesis and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve adipogenesis through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of lipid species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with and risk of developing T2D.

Keyword: insulin resistance

Dietary fatty acids modulate adipocyte TNFa production via regulation of its DNA promoter methylation levels.

The factors regulating TNF alpha (TNFa) levels could be considered therapeutic targets against metabolic syndrome development. DNA methylation is a potent regulator of gene expression and may be associated with protein levels. In this study we investigate whether the effect of dietary fatty acids on TNFa released from adipocytes might be associated with modifications of the TNFa promoter DNA methylation status. A group of rats was assigned to three diets with a different composition of saturated, monounsaturated and polyunsaturated fatty acids. Samples of visceral adipose tissues were taken for adipocyte isolation, in which released TNFa levels were measured, and for methylation and expression studies. In addition, 3 T3-L1 cells were treated with , oleic and linoleic acids, with and without 5-Azacitydine (5-AZA). After treatments, cells and supernatants were included in the same analyses as rat samples. TNFa promoter methylation levels, gene expression and secretion were different according to the diets and fatty treatments associated with them. Cells treated with 5-AZA displayed higher TNFa levels than in the absence of 5-AZA, without differences between fatty acids. According to our results, dietary fatty regulation of adipocyte TNFa levels may be mediated by epigenetic modifications of the TNFa promoter DNA methylation levels.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Mitochondrial dysfunction and inhibition of myoblast differentiation in mice with high-fat-diet-induced pre-diabetes.

Pre-diabetes is characterized by impaired glucose tolerance (IGT) and/or impaired fasting glucose. Impairment of skeletal muscle function is closely associated with the progression of diabetes. However, the entire pathological characteristics and mechanisms of pre-diabetes in skeletal muscle remain fully unknown. Here, we established a mouse model of pre-diabetes, in which 6-week-old male C57BL6/J mice were fed either normal diet or high-fat diet (HFD) for 8 or 16 weeks. Both non-fasting and fasting glucose levels and the results of glucose and tolerance tests showed that mice fed an 8-week HFD developed pre-diabetes with IGT; whereas mice fed a 16-week HFD presented with impaired fasting glucose and impaired glucose tolerance (IFG-IGT). Mice at both stages of pre-diabetes displayed decreased numbers of mitochondria in skeletal muscle. Moreover, IFG-IGT mice exhibited decreased mitochondrial membrane potential and ATP production in skeletal muscle and muscle degeneration characterized by a shift in muscle fibers from predominantly oxidative type I to glycolytic type II. Western blotting and histological analysis confirmed that myoblast differentiation was only inhibited in IFG-IGT mice. For primary skeletal muscle satellite cells, inhibition of differentiation was observed in -induced model. Moreover, enhanced myoblast differentiation increased glucose uptake and sensitivity. These findings indicate that pre-diabetes result in mitochondrial dysfunction and inhibition of myoblast differentiation in skeletal muscle. Therefore, interventions that enhance myoblast differentiation may improve of diabetes at the earlier stage.© 2018 Wiley Periodicals, Inc.

Keyword: insulin resistance

Clinopodium chinense Attenuates -Induced Vascular Endothelial Inflammation and through TLR4-Mediated NF- B and MAPK Pathways.

Elevated (PA) levels are associated with the development of inflammation, (IR) and endothelial dysfunction. Clinopodium chinense (Benth.) O. Kuntze has been shown to lower blood glucose and attenuate high glucose-induced vascular endothelial cells injury. In the present study we investigated the effects of ethyl acetate extract of C. chinense (CCE) on PA-induced inflammation and IR in the vascular endothelium and its molecular mechanism. We found that CCE significantly inhibited PA-induced toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs). Consequently, this led to the inhibition of the following downstream adapted proteins myeloid differentiation primary response gene 88, Toll/interleukin-1 receptor domain-containing adaptor-inducing interferon- and TNF receptor-associated factor 6. Moreover, CCE inhibited the phosphorylation of Ikappa B kinase , nuclear factor kappa-B (NF- B), c-Jun N-terminal kinase, extracellular regulated protein kinases, p38-mitogen-activated protein kinase (MAPK) and subsequently suppressed the release of tumor necrosis factor- , interleukin-1 (IL-1 ) and IL-6. CCE also inhibited IRS-1 serine phosphorylation and ameliorated -mediated tyrosine phosphorylation of IRS-1. Moreover, CCE restored serine/threonine kinase and endothelial nitric oxide synthase (eNOS) activation and thus increased -mediated nitric oxide (NO) production in PA-treated HUVECs. This led to reverse mediated endothelium-dependent relaxation, eNOS phosphorylation and NO production in PA-treated rat thoracic aortas. These results suggest that CCE can significantly inhibit the inflammatory response and alleviate impaired signaling in the vascular endothelium by suppressing TLR4-mediated NF- B and MAPK pathways. Therefore, CCE can be considered as a potential therapeutic candidate for endothelial dysfunction associated with IR and diabetes.

Keyword: insulin resistance

Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.

Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves liver injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 diabetes mellitus (T2DM) and to determine the underlying mechanisms of these effects.A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with . Major signalling profiles involved in fatty oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry.Nuciferine restored impaired glucose tolerance and in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed -induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation.Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.© 2018 The British Pharmacological Society.

Keyword: insulin resistance

Losartan Improves Palmitate-Induced in 3T3-L1 Adipocytes Through Upregulation of Src Phosphorylation.

Angiotensin II type 1 receptor blocker losartan has shown strongly anti- properties and ; however, the underlying mechanisms are poorly understood. In this study, we demonstrate that losartan administration increased phosphorylation of Akt and its downstream Akt substrate of 160\u2009kDa (AS160), enhanced plasma membrane translocation of glucose transporter type 4 (GLUT4), and increased glucose uptake, along with increased Src phosphorylation as well as reduced expression of docking protein 1(DOK1) in palmitate-treated 3T3-L1 adipocytes. The beneficial impacts of losartan on signaling were diminished in Src-deficient 3T3-L1 adipocytes. In addition, suppressed expression of DOK1 by losartan was abolished by Src knockdown. Our results suggest that anti- ability of losartan is mediated by Src/DOK1/Akt pathway.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: insulin resistance

The effect of enterolactone on sphingolipid pathway and hepatic development in HepG2 cells.

Obesity and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of -stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: insulin resistance

Sphingomyelinase Down-regulation Alleviates Vascular Endothelial in Diabetic Rats.

in endothelial cells contributes to the development of cardiovascular disease in patients with type 2 diabetes. sphingomyelinase (ASM) is a soluble glycoprotein which plays a vital role in the development and progression of various diseases such as cardiovascular and metabolic diseases. However, it remains unknown if ASM regulates in vascular endothelial cells in type 2 diabetes. ASM down-regulation with gene silencing and selective inhibitor amitriptyline was used in the rat aortic endothelial cells (RAECs) treated with (PA), a common saturated free fatty , which is thought to be the major cause of . It was shown that ASM down-regulation increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of pIRS-1-ser307 and AKT-ser473 via ceramide, consequently resulting in the decrease of the production of endothelial nitric oxide synthase (eNOS) and nitric oxide in PA-induced RAECs. We further found that ASM down-regulation blocked the Nox2- and Nox4-dependent superoxide (O ) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vivo, amitriptyline relieved the vasodilatory response to acetylcholine and restored the level of ceramide, Nox2 and Nox4 in the aorta endothelium of high-fat diet-fed rats following an injection of streptozotocin. Taken together, these results suggest that ASM down-regulation can improve endothelial which is attributed to inhibiting redox signalling in RAECs. Thus, these data support the idea that ASM is a promising clinical biomarker and potential therapeutic target for diabetic vascular complication.© 2018 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Keyword: insulin resistance

Dimethylarginine Dimethylaminohydrolase 1 Protects Against High-Fat Diet-Induced Hepatic Steatosis and in Mice.

High plasma concentrations of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, are associated with hepatic dysfunction in patients with nonalcoholic fatty liver disease (NAFLD). However, it is unknown whether ADMA is involved in the pathogenesis of NAFLD. Dimethylarginine dimethylaminohydrolase 1 (DDAH1) is an enzyme that degrades ADMA. In this study, we used Ddah1 mice to investigate the effects of the ADMA/DDAH1 pathway on high-fat diet (HFD)-induced hepatic steatosis.After HFD feeding for 20 weeks, Ddah1 mice were more obese and had developed more severe hepatic steatosis and worse compared with wild-type (WT) mice. In the livers of HFD-fed mice, loss of DDAH1 resulted in higher levels of lipogenic genes, lower expression of β-oxidation genes, and greater induction of oxidative stress, endoplasmic reticulum stress, and inflammation than in the WT livers. Furthermore, ADMA treatment in HepG2 cells led to oxidative stress and steatosis, whereas overexpression of DDAH1 attenuated -induced steatosis, oxidative stress, and inflammation. Innovation and Conclusion: Our results provide the first direct evidence that the ADMA/DDAH1 pathway has a marked effect on hepatic lipogenesis and steatosis induced by HFD feeding. Our findings suggest that strategies to increase DDAH1 activity in hepatocytes may provide a novel approach to attenuate NAFLD development. Antioxid. Redox Signal. 26, 598-609.

Keyword: insulin resistance

Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as , dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut microbiota, and hepatic metabolism.

Keyword: insulin resistance

Decreased lipid metabolism but increased FA biosynthesis are coupled with changes in liver microRNAs in obese subjects with NAFLD.

Many controversies regarding the association of liver miRNAs with obesity and nonalcoholic fatty liver diseases (NAFLD) call for additional validations. This study sought to investigate variations in genes and hepatic miRNAs in a sample of obese patients with or without NAFLD and human hepatocytes (HH).A total of 60 non-consecutive obese women following bariatric surgery were recruited. Subjects were classified as NAFLD (n=17), borderline (n=24) and controls (n=19) with normal enzymatic profile, liver histology and ultrasound assessments. Profiling of 744 miRNAs was performed in 8 obese women with no sign of hepatic disease and 11 NAFLD patients. Additional validation and expression of genes related to de novo fatty (FA) biosynthesis, uptake, transport and β-oxidation; glucose metabolism, and inflammation was tested in the extended sample. Induction of NAFLD-related genes and miRNAs was examined in HepG2 cells and primary HH treated with (PA), a combination of palmitate and oleic , or high glucose, and (HG) mimicking in NAFLD.In the discovery sample, 14 miRNAs were associated with NAFLD. Analyses in the extended sample confirmed decreased miR-139-5p, miR-30b-5p, miR-122-5p and miR-422a, and increased miR-146b-5p in obese subjects with NAFLD. Multiple linear regression analyses disclosed that NAFLD contributed independently to explain miR-139-5p (P=0.005), miR-30b-5p (P=0.005), miR-122-5p (P=0.021), miR-422a (P=0.007) and miR-146a (P=0.033) expression variance after controlling for confounders. Decreased miR-122-5p in liver was associated with impaired FA usage. Expression of inflammatory and macrophage-related genes was opposite to decreased miR-30b-5p, miR-139-5p and miR-422a, whereas increased miR-146b-5p was associated with FABP4 and decreased glucose metabolism and FA mobilization. In partial agreement, PA (but not HG) led to decreased miR-139-5p, miR-30b-5p, miR-422a and miR-146a in vitro, in parallel with increased lipogenesis and FA transport, decreased glucose metabolism and diminished FA oxidation.This study confirms decreased liver glucose and lipid metabolism but increased FA biosynthesis coupled with changes in five unique miRNAs in obese patients with NAFLD.

Keyword: insulin resistance

5-LO inhibition ameliorates -induced ER stress, oxidative stress and via AMPK activation in murine myotubes.

Leukotriene B4 (LTB4) production via the 5-lipoxygenase (5-LO) pathway contributes to the development of in adipose and hepatic tissues, but the role of LTB4 in skeletal muscle is relatively unknown. Here, the authors investigated the role of LTB4 in C2C12 myotubes in (PA)-induced ER stress, inflammation and . PA (750\u2009μM) evoked lipotoxicity (ER stress, oxidative stress, inflammation and ) in association with LTB4 production. 5-LO inhibition reduced all the lipotoxic effects induced by PA. On the other hand, PA did not induce cysteinyl leukotrienes (CysLTs), which themselves had no effect on ER stress and inflammation. The beneficial effects of 5-LO suppression from PA-induced lipotoxicity were related with AMPK activation. In ob/ob mice, once daily oral administration of zileuton (50, 100\u2009mg/kg) for 5 weeks improved , increased AMPK phosphorylation, and reduced LTB4 and ER stress marker expression in skeletal muscle. These results show that 5-LO inhibition by either zileuton or 5-LO siRNA protects C2C12 myotubes from PA-induced lipotoxicity, at least partly via AMPK activation, and suggest that the in vivo -sensitizing effects of zileuton are in part attributable to its direct action on skeletal muscle via LTB4 downregulation followed by AMPK activation.

Keyword: insulin resistance

Disruption of the mitochondria-associated ER membrane (MAM) plays a central role in -induced .

The mitochondria-associated ER membrane (MAM) is a specialized subdomain of ER that physically connects with mitochondria. Although disruption of inter-organellar crosstalk via the MAM impairs cellular homeostasis, its pathological significance in in type 2 diabetes mellitus remains unclear. Here, we reveal the importance of reduced MAM formation in the induction of fatty -evoked in hepatocytes. (PA) repressed -stimulated Akt phosphorylation in HepG2 cells within 12h. Treatment with an inhibitor of the ER stress response failed to restore PA-mediated suppression of Akt activation. Mitochondrial reactive oxygen species (ROS) production did not increase in PA-treated cells. Even short-term exposure (3h) to PA reduced the calcium flux from ER to mitochondria, followed by a significant decrease in MAM contact area, suggesting that PA suppressed the functional interaction between ER and mitochondria. Forced expression of mitofusin-2, a critical component of the MAM, partially restored MAM contact area and ameliorated the PA-elicited suppression of sensitivity with Ser473 phosphorylation of Akt selectively improved. These results suggest that loss of proximity between ER and mitochondria, but not perturbation of homeostasis in the two organelles individually, plays crucial roles in PA-evoked Akt inactivation in hepatic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Liraglutide ameliorates palmitate-induced through inhibiting the IRS-1 serine phosphorylation in mouse skeletal muscle cells.

A reduction in -stimulated glucose uptake in skeletal muscles is a characteristic of and type 2 diabetes mellitus (T2DM). The glucagon-like peptide (GLP)-1 agonist liraglutide can reduce blood glucose levels in individuals with T2DM. However, its effect on -induced glucose metabolism in the skeletal muscle of is unknown. We investigated the effects and action mechanisms of liraglutide on (IR) in the\xa0skeletal muscle cells treatment with (PA).The cell-surface GLUT4myc levels were determined by an antibody-coupled colorimetric assay. The phosphorylation levels of Akt, PI3K(p85α), AS160, IRS1, IKK, and JNK were determined by western blotting. The quantifications of mRNA levels of TNFα, IL-1β, and IL-6 were determined by real-time PCR. Analysis of variance was used for data analysis.PA elevated not only phosphorylation of JNK, IRS1 serines, and IKKα/β, but also the expression of IL-6,\xa0TNFα and IL-1β in C2C12-GLUT4myc cells. PA can reduce phosphorylation of IRS1 tyrosine. These effects of PA were reversed by liraglutide. In addition, liraglutide can reverse PA-decreased -stimulated cell-surface GLUT4 levels, Akt, PI3K(p85α), and AS160 phosphorylation.Liraglutide can enhance -induced GLUT4 translocation by inhibiting IRS1 serine phosphorylation in PA-treated muscle cells.

Keyword: insulin resistance

Puerarin acts on the skeletal muscle to improve sensitivity in diabetic rats involving μ-opioid receptor.

Puerarin, a major active isoflavone extracted from the root of Pueraria lobate, significantly increases plasma β-endorphin and levels and improves impaired signaling in diabetic animals. However, the target tissues and underlying mechanisms in and through which puerarin functions to ameliorating remains largely unclear. In this study, we showed that puerarin enhanced μ-opioid receptor expression and phosphorylation, and increased -stimulated glucose transporter 4 translocation to the plasma membrane in the skeletal muscle of diabetic rats, which were recaptured by a direct application of puerarin in the palmitate-induced -resistant L6 myotubes. Naloxone, an antagonist of μ-opioid receptor, blocked these functions of puerarin. No β-endorphin was detected either in the muscle of diabetic rats or in the palmitate-induced -resistant L6 cells. Furthermore, we presented the evidence to show the interaction between μ-opioid receptor and receptor substrate 1 in the muscle tissues and cells. These results suggested that puerarin improved sensitivity in the skeletal muscle at least in part by its local effects involving μ-opioid receptor function.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

Obesity increases the risk of developing diabetes mellitus. Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in diabetes with obesity. Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II diabetes. Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling pathways was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of diabetes with obesity that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for diabetes with obesity.

Keyword: insulin resistance

Attenuation of palmitate induced in muscle cells by harmala, clove and river red gum.

The present study aimed to decipher the mechanism of action of selected anti-diabetic plants extracts on mediated in muscle cells. Our results showed that extract from Peganum harmala seeds, Eucalyptus camaldulensis and Syzygium aromaticum leaves, showed significant antioxidant activity. We found that these extracts were able to affect stress signalling by reducing p-38 MAP kinase phosphorylation. They also reduced phosphorylation of substrate for receptor (IRS) at serine residues and increased its phosphorylation at tyrosine residues and also enhanced PKB phosphorylation. Glucose uptake was also enhanced in muscle cells after treatment with these extracts. Extracts from Lantana camara, Psidium gujava fruit and different parts of Cassia alata did not affect FFA mediated down-regulation of signalling. The study conclude that seeds of Peganum harmala and leaves of Eucalyptus camaldulensis and Syzygium aromaticum enhanced signal transduction and glucose uptake in muscle cells via reducing oxidative stress. As a result, these herbal extracts may be considered useful to protect from .

Keyword: insulin resistance

Metabolomic change due to combined treatment with myo-inositol, D-chiro-inositol and glucomannan in polycystic ovarian syndrome patients: a pilot study.

Polycystic ovarian syndrome (PCOS) is a highly variable syndrome and one of the most common female endocrine disorders. Although the association inositols-glucomannan may represent a good therapeutic strategy in the treatment of PCOS women with , the effect of inositols on the metabolomic profile of these women has not been described yet.Fifteen PCOS-patients and 15 controls were enrolled. Patients were treated with myo-inositol (1.75\u2009g/day), D-chiro-inositol (0.25\u2009g/day) and glucomannan (4\u2009g/day) for 3\u2009months. Blood concentrations of glucose, , triglycerides and cholesterol, and ovary volumes and antral follicles count, as well as metabolomic profiles, were evaluated for control subjects and for cases before and after treatment. PCOS-patients had higher BMI compared with Controls, BMI decreased significantly after 3\u2009months of treatment although it remained significantly higher compared to controls. 3-methyl-1-hydroxybutyl-thiamine-diphosphate, valine, phenylalanine, ketoisocapric, linoleic, lactic, glyceric, citric and , glucose, glutamine, creatinine, arginine, choline and tocopherol emerged as the most relevant metabolites for distinguishing cases from controls.Our pilot study has identified a complex network of serum molecules that appear to be correlated with PCOS, and with a combined treatment with inositols and glucomannan.ClinicalTial.gov, . Registered 1st August 2018 - Retrospectively registered, .

Keyword: insulin resistance

Egr2 enhances via JAK2/STAT3/SOCS-1 pathway in HepG2 cells treated with palmitate.

is generally responsible for the pathogenesis of type 2 diabetes mellitus (T2DM). Early growth response proteins-2 (Egr2) has been reported to be able to increase the expression of the suppressors of cytokine signaling-1 (SOCS-1), and impair signaling pathway through suppression of receptor substrates (IRS), including IRS-1 and IRS-2. However, whether Egr2 is directly involved in the development of , and how its potential contributions to still remain unknown. Here, our present investigation found that the expression levels of Egr2 were up-regulated when occurs, and knockdown of Egr2 abolished the effect of in HepG2 cells induced with palmitate (PA). Importantly, inhibition of Egr2 decreased the expression of SOCS-1 as well as reduced phosphorylation of JAK2 and STAT3. And, our data indicated that silencing of Egr2 accelerated hepatic glucose uptake and reversed the impaired lipid metabolism upon . In summary, the present study confirms that Egr2 could deteriorate via the pathway of JAK2/STAT3/SOCS-1 and may shed light on resolving and further the pathogenesis of T2DM.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Long-term exposition to a high fat diet favors the appearance of β-amyloid depositions in the brain of C57BL/6J mice. A potential model of sporadic Alzheimer\'s disease.

The sporadic and late-onset form of Alzheimer\'s disease (AD) constitutes the most common form of dementia. This non-familiar form could be a consequence of metabolic syndrome, characterized by obesity and the development of a brain-specific known as type III diabetes. This work demonstrates the development of a significant AD-like neuropathology due to these metabolic alterations.C57BL/6J mice strain were divided into two groups, one fed with a diet rich in (high-fat diet, HFD) since their weaning until 16 months of age, and another group used as a control with a regular diet. The analyses were carried out in the dentate gyrus area of the hippocampus using a Thioflavin-S stain and immunofluorescence assays.The most significant finding of the present research was that HFD induced the deposition of the βA peptide. Moreover, the diet also caused alterations in different cell processes, such as increased inflammatory reactions that lead to a decrease in the neuronal precursor cells. In addition, the results show that there were also dysregulations in normal autophagy and apoptosis, mechanisms related to βA formation.The present findings confirm that HFD favors the formation of βA depositions in the brain, a key feature of AD, supporting the metabolic hypothesis of sporadic AD.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: insulin resistance

PAQR3 regulates phosphorylation of FoxO1 in -resistant HepG2 cells via NF-κB signaling pathway.

is a significant feature of type 2 diabetes mellitus and glucose and lipid metabolism disorders. Activation of NF-κB signaling pathway plays an important role in the formation of . FoxO1 plays a major role in regulating glucose and lipid metabolism, as well as signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under . In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced -resistant HepG2 cells, thereby causing glucose and lipid metabolism disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced -resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and lipid metabolism disorders induced by .Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Brain impairs hippocampal synaptic plasticity and memory by increasing GluA1 palmitoylation through FoxO3a.

High-fat diet (HFD) and metabolic diseases cause detrimental effects on hippocampal synaptic plasticity, learning, and memory through molecular mechanisms still poorly understood. Here, we demonstrate that HFD increases deposition in the hippocampus and induces hippocampal leading to FoxO3a-mediated overexpression of the palmitoyltransferase zDHHC3. The excess of along with higher zDHHC3 levels causes hyper-palmitoylation of AMPA glutamate receptor subunit GluA1, hindering its activity-dependent trafficking to the plasma membrane. Accordingly, AMPAR current amplitudes and, more importantly, their potentiation underlying synaptic plasticity were inhibited, as well as hippocampal-dependent memory. Hippocampus-specific silencing of Zdhhc3 and, interestingly enough, intranasal injection of the palmitoyltransferase inhibitor, 2-bromopalmitate, counteract GluA1 hyper-palmitoylation and restore synaptic plasticity and memory in HFD mice. Our data reveal a key role of FoxO3a/Zdhhc3/GluA1 axis in the HFD-dependent impairment of cognitive function and identify a novel mechanism underlying the cross talk between metabolic and cognitive disorders.

Keyword: insulin resistance

Anti-diabetic effects and mechanisms of action of a Chinese herbal medicine preparation JQ-R and in diabetic KK mice.

Refined-JQ (JQ-R) is a mixture of refined extracts from (Ranunculaceae), (Leguminosae) and (Caprifoliaceae), the three major herbs of JinQi-JiangTang tablet, a traditional Chinese medicine (TCM) formula. The mechanisms by which JQ-R regulates glucose metabolism and improves sensitivity were studied in type 2 diabetic KK mice and -resistant L6 myotubes. To investigate the mechanisms by which JQ-R improves sensitivity, a model of -resistant cells induced with (PA) was established in L6 myotubes. Glucose uptake and expression of factors involved in signaling, stress, and inflammatory pathways were detected by immunoblotting. JQ-R showed beneficial effects on glucose homeostasis and in a euglycemic clamp experiment and decreased fasting levels in diabetic KK mice. JQ-R also improved the plasma lipid profiles. JQ-R directly increased the activity of superoxide dismutase (SOD) and decreased malondialdehyde (MDA) as well as inducible nitric oxide synthase (iNOS) levels in -resistant L6 cells, and elevated the -stimulated glucose uptake with upregulated phosphorylation of AKT. The phosphorylation levels of nuclear factor kappa B (NF-B p65), inhibitor of NF-B (IB ), c-Jun N-terminal kinase (JNK1/2) and extracellular-signal-regulated kinases (ERK1/2) were also changed after JQ-R treatment compared with the control group. Together these findings suggest that JQ-R improved glucose and lipid metabolism in diabetic KK mice. JQ-R directly enhanced -stimulated glucose uptake in -resistant myotubes with improved signalling and inflammatory response and oxidative stress. JQ-R could be a candidate to achieve improved glucose metabolism and sensitivity in type 2 diabetes mellitus.

Keyword: insulin resistance

Neutral ceramidase-enriched exosomes prevent -induced in H4IIEC3 hepatocytes.

Oversupply of free fatty acids such as (PA) from the portal vein may cause liver . Production of reactive oxygen species plays a pivotal role in PA-induced in H4IIEC3 hepatocytes. Recently, we found that exosomes secreted from INS-1 cells that were transfected with neutral ceramidase (NCDase) plasmids had raised NCDase activity; these NCDase-enriched exosomes could inhibit PA-induced INS-1 cell apoptosis. Here, we showed that PA reduced -stimulated tyrosine phosphorylation of receptor substrate 2 and decreased -stimulated uptake of the fluorescent glucose analog 2-NBDG, confirming that occurred in PA-treated H4IIEC3 cells. Moreover, NCDase-enriched exosomes from INS-1 cells rescued PA-induced H4IIEC3 and blocked PA-induced reactive oxygen species production in which ceramide was involved.

Keyword: insulin resistance

Hemin Improves Sensitivity and Lipid Metabolism in Cultured Hepatocytes and Mice Fed a High-Fat Diet.

Hemin is a breakdown product of hemoglobin. It has been reported that the injection of hemin improves lipid metabolism and sensitivity in various genetic models. However, the effect of hemin supplementation in food on lipid metabolism and sensitivity is still unclear, and whether hemin directly affects cellular sensitivity is yet to be elucidated. Here we show that hemin enhances -induced phosphorylation of receptors, Akt, Gsk3β, FoxO1 and cytoplasmic translocation of FoxO1 in cultured primary hepatocytes under -resistant conditions. Furthermore, hemin diminishes the accumulation of triglyceride and increases in free fatty content in primary hepatocytes induced by palmitate. Oral administration of hemin decreases body weight, energy intake, blood glucose and triglyceride levels, and improves and glucose tolerance as well as hepatic signaling and hepatic steatosis in male mice fed a high-fat diet. In addition, hemin treatment decreases the mRNA and protein levels of some hepatic genes involved in lipogenic regulation, fatty synthesis and storage, and increases the mRNA level and enzyme activity of CPT1 involved in fatty oxidation. These data demonstrate that hemin can improve lipid metabolism and sensitivity in both cultured hepatocytes and mice fed a high-fat diet, and show the potential beneficial effects of hemin from food on lipid and glucose metabolism.

Keyword: insulin resistance

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the signaling pathway, such as mTOR, Akt, IRS-1, and the receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: insulin resistance

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of sensitivity. In this study, the impact of SA on sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for ) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: insulin resistance

Fatty synthesis configures the plasma membrane for inflammation in diabetes.

Dietary fat promotes pathological through chronic inflammation. The inactivation of inflammatory proteins produced by macrophages improves diet-induced diabetes, but how nutrient-dense diets induce diabetes is unknown. Membrane lipids affect the innate immune response, which requires domains that influence high-fat-diet-induced chronic inflammation and alter cell function based on phospholipid composition. Endogenous fatty synthesis, mediated by fatty synthase (FAS), affects membrane composition. Here we show that macrophage FAS is indispensable for diet-induced inflammation. Deleting Fasn in macrophages prevents diet-induced , recruitment of macrophages to adipose tissue and chronic inflammation in mice. We found that FAS deficiency alters membrane order and composition, impairing the retention of plasma membrane cholesterol and disrupting Rho GTPase trafficking-a process required for cell adhesion, migration and activation. Expression of a constitutively active Rho GTPase, however, restored inflammatory signalling. Exogenous palmitate was partitioned to different pools from endogenous lipids and did not rescue inflammatory signalling. However, exogenous cholesterol, as well as other planar sterols, did rescue signalling, with cholesterol restoring FAS-induced perturbations in membrane order. Our results show that the production of endogenous fat in macrophages is necessary for the development of exogenous-fat-induced through the creation of a receptive environment at the plasma membrane for the assembly of cholesterol-dependent signalling networks.

Keyword: insulin resistance

Risk of diabetes associated with fatty acids in the de novo lipogenesis pathway is independent of sensitivity and response: the Atherosclerosis Study (IRAS).

To examine the associations of fatty acids in the de novo lipogenesis (DNL) pathway, specifically myristic (14:0), (16:0), palmitoleic (c16:1\u2009n-7), myristoleic (c14:1n5), stearic (18:0) and oleic (c18:1\u2009n-9), with 5-year risk of type 2 diabetes. We hypothesized that DNL fatty acids are associated with risk of type 2 diabetes independent of sensitivity.We evaluated 719 (mean age 55.1±8.5 years, 44.2% men, 42.3% Caucasians) participants from the Atherosclerosis Study. Multivariable logistic regression models with and without adjustment of sensitivity were used to assess prospective associations of DNL fatty acids with incident type 2 diabetes.Type 2 diabetes incidence was 20.3% over 5\u2009years. In multivariable regression models, , palmitoleic, myristic, myristoleic and oleic acids were associated with increased risk of type 2 diabetes (p<0.05). had the strongest association (OR per standard unit of 1.46; 95%\u2009CI 1.23 to 1.76; p<0.001), which remained similar with addition of sensitivity and acute response (AIR) to the model (OR 1.36; 95%\u2009CI 1.09 to 1.70, p=0.01). Oleic and palmitoleic acids were also independently associated with incident type 2 diabetes. In multivariable models, ratios of fatty acids corresponding to stearoyl CoA desaturase-1 and Elovl6 enzymatic activity were significantly associated with risk of type 2 diabetes independent of sensitivity and AIR.We observed associations of DNL fatty acids with type 2 diabetes incidence independent of sensitivity.

Keyword: insulin resistance

Intracellular and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes.

Type 2 diabetes mellitus is characterized by . However, the complete molecular mechanism remains unclear. In this study, zebrafish were fed a long-term high-fat diet to induce type 2 diabetes, which resulted in a higher body weight, body mass index, more lipid vacuoles in liver, increased transcription level in liver, brain and muscle, and high fasting blood glucose in the high-fat diet zebrafish. Oppositely, the transcription levels of substrate-2 and glucose transporter 2 were significantly decreased, indicating signaling pathway and glucose transport impaired in the -targeting tissues. Transcription of the autophagy-related genes, ATG3, ATG4B, ATG5, ATG7, ATG12, and FOXO3, were decreased but autophagy inhibitor gene m-TOR increased, and autophagy-flux was inhibited in liver of the high-fat diet zebrafish. Main of these changes were confirmed in -treated HepG2 cells. Further, in co-immunoprecipitation and subcellular co-localization experiments, the conjunction of preproinsulin with cargo-recognition protein p62 increased, but conjuncts of autophagosome with p62-cargo, lysosomes with p62-cargo, and autolysosomes decreased apparently. Interestingly, lysosomes, autolysosomes and conjuncts of p62- localized at the periphery of -treated cells, the margination of lysosomes may mediate deactivation of proteases activity. These findings suggest that intracellular high-lipid may trigger defective autophagy, defective downstream signaling of and accumulated intracellular preproinsulin, leading to dysregulation of cell homeostasis mechanism, which may be one of reasons involved in in type 2 diabetes.

Keyword: insulin resistance

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, , and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low fat diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic lipid accumulation and lowered indices of . Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and diabetes, and (3) dietary SFA type impacts LCPUFA metabolism.Published by Elsevier Inc.

Keyword: insulin resistance

Metabolic pathways of oleic and are intensified in PCOS patients with normal androgen levels.

The aetiology and pathogenesis polycystic ovary syndrome (PCOS) remain uncertain and thus the relative studies are still crucial.Our aim was to analyse the fatty acids profiles of the main phospholipids species in serum from women with PCOS classified into phenotypes, and to diagnose women more susceptible to the occurrence of inflammatory state.PCOS screening tests were performed in The Clinic of Gynecology and Urogynecology of Pomeranian Medical University in the 2014-2015 years.The study are designed for general community and a primary care or referral center.39 patients with PCOS, diagnosed according to Rotterdam\'s criteria, and 14 healthy women, as a control group, participated in this study. Fatty profiles were investigated using gas chromatography. A total of 36 fatty acids and their derivatives were identified and quantified.Changes in fatty acids profile in plasma from women with PCOS phenotypes are not identical.The analyses showed lowered level of total SFA, increase in the concentration of caprylic and the activation of and oleic acids pathways. The level of nervonic was several times higher than in the control group, and the levels of behenic and tricosanoic acids were reduced.In both phenotypes the alternative metabolic pathways of oleic were activated, but they were more pronounced in women with proper level of androgens. Gamma-linolenic (C18:3n6) can be a factor protecting hyperandrogenic women.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

Depot-specific differences in fatty composition and distinct associations with lipogenic gene expression in abdominal adipose tissue of obese women.

Cardiometabolic diseases are primarily linked to enlarged visceral adipose tissue (VAT). However, some data suggest heterogeneity within the subcutaneous adipose tissue (SAT) depot with potential metabolic differences between the superficial SAT (sSAT) and deep SAT (dSAT) compartments. We aimed to investigate the heterogeneity of these three depots with regard to fatty (FA) composition and gene expression. Adipose tissue biopsies were collected from 75 obese women undergoing laparoscopic gastric bypass surgery. FA composition and gene expression were determined with gas chromatography and quantitative real-time-PCR, respectively. Stearoyl CoA desaturase-1 (SCD-1) activity was estimated by product-to-precursor FA ratios. All polyunsaturated FAs (PUFA) with 20 carbons were consistently lower in VAT than either SAT depots, whereas essential PUFA (linoleic , 18:2n-6 and α-linolenic , 18:3n-3) were similar between all three depots. Lauric and were higher and lower in VAT, respectively. The SCD-1 product palmitoleic as well as estimated SCD-1 activity was higher in VAT than SAT. Overall, there was a distinct association pattern between lipid metabolizing genes and individual FAs in VAT. In conclusion, SAT and VAT are two distinct depots with regard to FA composition and expression of key lipogenic genes. However, the small differences between sSAT and dSAT suggest that FA metabolism of SAT is rather homogenous.

Keyword: insulin resistance

Advanced Liver Fibrosis Is Independently Associated with and Levels in Patients with Non-Alcoholic Fatty Liver Disease.

Changes in lipid metabolism occur during the development and progression non-alcoholic fatty liver disease (NAFLD). However, the fatty (FA) profile in red blood cells (RBC) from patients with liver fibrosis remains unexplored. Thus, the goal of this study was to evaluate the fatty profile in RBC, dietary lipid intake and indicators in patients with NAFLD, according to the degree of hepatic fibrosis. Using elastography, patients were classified with ( = 52) and without ( = 37) advanced liver fibrosis. The fatty profile in RBC was analyzed using gas chromatography and the lipid intake was evaluated through a 24-h dietary recall. Subjects with advanced liver fibrosis had higher levels of , stearic and oleic and total monounsaturated fatty (MUFA) and ( < 0.05), and lower levels of elongase very long chain fatty acids protein-6 and the delta-5-desaturase enzymatic activity ( < 0.05). These results suggest a lack of regulation of enzymes related to FA metabolism in patients with advanced fibrosis.

Keyword: insulin resistance

Vernonia amygdalina Delile extract inhibits the hepatic gluconeogenesis through the activation of adenosine-5\'monophosph kinase.

It has been reported that Vernonia amygdalina Delile(VA) presents an anti-diabetic effect, and the effect of VA on lowering glucose is formulated via suppressing the expression of the key hepatic gluconeogenesis enzyme. Therefore, we further explored the probable mechanism of VA on dismissing hepatic gluconeogenesis through the activation of adenosine-5\' monophosphate kinase (AMPK) in vivo and in vitro.We developed type 2 diabetic mice with STZ and oral administration with VA (50\u202fmg/kg, 100\u202fmg/kg and 150\u202fmg/kg) once a day for 6 weeks. Fasting blood glucose (FBG), fasting (FINS) and oral glucose tolerance tests (OGTT) were conducted. The expression levels of AMPK, phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase) proteins in live were evaluated by western blot. Then, we further explored the mechanism of VA on hepatic gluconeogenesis in vitro experiments. Glucose production and the expression of AMPK, PEPCK and G6Pase proteins were detected after VA treatment with the presence of the AMPK inhibitor Compound C.VA reduced FBG and caused a significant improvement in glucose tolerance and (HOMA-IR) in STZ-induced mice. VA inhibited the elevated expression of gluconeogenesis key enzymes (PEPCK and G6Pase) and up-regulated AMPK activity in liver. In (PA)-induced HepG2 cells, VA decreased glucose production and the expression of PEPCK and G6Pase proteins, also activated AMPK pathway. The effects of VA on gluconeogenesis could be reversed by Compound C.These results reveal that VA suppresses hepatic gluconeogenesis at least partially through activating the AMPK.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: insulin resistance

d- chiro-Inositol Ameliorates High Fat Diet-Induced Hepatic Steatosis and via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate . However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high fat diet and saturated -treated hepatocytes. DCI attenuated free fatty uptake by the liver via lipid trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced . These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces lipid deposition, and ameliorates via regulation of PKCε-PI3K/AKT axis.

Keyword: insulin resistance

Derivatization enhanced separation and sensitivity of long chain-free fatty acids: Application to asthma using targeted and non-targeted liquid chromatography-mass spectrometry approach.

Long chain-free fatty acids (LCFFAs) play pivotal roles in various physiological functions, like inflammation, , hypertension, immune cell behavior and other biological activities. However, the detection is obstructed by the low contents, structural diversity, high structural similarity, and matrix interference. Herein, a fast cholamine-derivatization, within 1\xa0min at room temperature, coupled with liquid chromatography-mass spectrometry (LC-MS) approach was developed to determine LCFFAs in complex samples. After derivatization, the ionization and separation efficiency were significantly improved, which resulted in up to 2000-fold increase of sensitivity compared with non-derivatization method, and the limits of detection were at low femtogram level. As well, this approach was applied successfully in the rapid profiling or quantification of targeted and non-targeted LCFFAs in the sera of healthy human and asthma patients. The targeted metabolomics method showed that the contents of 17 PUFAs were significantly changed in asthma patients, especially hydroxyeicosatetraenoic acids (HETEs), hydroperoxyeicosatetraenoic (HPETEs) and prostaglandins (PGs). The non-targeted method resulted in the tentatively identification of 35 LCFFAs including 31 saturated and mono-unsaturated LCFFAs, and 4 bile acids, except for 27 poly-unsaturated fatty acids (PUFAs), and the multivariate analysis indicated that eicosapentaenoic (EPA), ursodeoxycholic , deoxycholic , isodeoxycholic , , 2-lauroleic and lauric also have significant difference between healthy and asthma groups except for 17 PUFAs. To the best of our knowledge, this is the first report on the relationship of asthma with 5(S)-, 15(S)-HPETE, 8(S)-, 11(S)-HETE, 15(S)-HEPE, PGA2, PGB2, PGE1, PGF1α, PGJ2, and 13, 14-dehydro-15-keto PGF2α (DK-PGF2α).Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the liver was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in secretion and . The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and obesity in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

Keyword: insulin resistance

Nucleophosmin/B23 contributes to hepatic through the modulation of NF-κB pathway.

Nucleophosmin (NPM)/B23 is an important nucleolar phosphoprotein involved in the regulation of assorted cellular signaling pathways. In the present study, we revealed a critical role of NPM in liver . NPM is markedly upregulated in -resistant liver tissues and (PA)-exposed HepG2 cells both at mRNA and protein levels. Ectopic expression of NPM in hepatocytes aggravated PA-induced , lipid droplet accumulation, glucose intake impairment as well as the expression of gluconeogenic genes. Coinciding with these results, interference of NPM using small interfering RNA (siRNA) oligos ameliorated PA-induced , as revealed by increased phosphorylation of AKT and GSK3β following treatment. As predicted, PA-triggered alterations in glucose intake and the expression of gluconeogenic enzymes were attenuated following NPM depletion. Finally, we showed that NPM plays an indispensible role in PA-induced activation of NF-κB pathway. Both of NF-κB p65 phosphorylation and nuclear translocation were impeded by NPM interference in PA-treated HepG2 cells. Taken together, these findings explicitly demonstrate that NPM participates in the development of liver , suggesting that NPM may serve as a potential therapeutic target of type 2 diabetes.Copyright © 2019. Published by Elsevier Inc.

Keyword: insulin resistance

The effect of on inflammatory response in macrophages: an overview of molecular mechanisms.

is a saturated fatty whose blood concentration is elevated in obese patients. This causes inflammatory responses, where toll-like receptors (TLR), TLR2 and TLR4, play an important role. Nevertheless, is not only a TLR agonist. In the cell, this fatty is converted into phospholipids, diacylglycerol and ceramides. They trigger the activation of various signaling pathways that are common for LPS-mediated TLR4 activation. In particular, metabolic products of affect the activation of various PKCs, ER stress and cause an increase in ROS generation. Thanks to this, also strengthens the TLR4-induced signaling. In this review, we discuss the mechanisms of inflammatory response induced by . In particular, we focus on describing its effect on ER stress and IRE1α, and the mechanisms of NF-κB activation. We also present the mechanisms of inflammasome NLRP3 activation and the effect of on enhanced inflammatory response by increasing the expression of FABP4/aP2. Finally, we focus on the consequences of inflammatory responses, in particular, the effect of TNF-α, IL-1β and IL-6 on . Due to the high importance of macrophages and the production of proinflammatory cytokines by them, this work mainly focuses on these cells.

Keyword: insulin resistance

Attenuation of Free Fatty -Induced Muscle by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle action leading to and Type 2 diabetes mellitus. Serine phosphorylation of the receptor substrate (IRS) is linked to and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract . We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle has never been examined. In the current study, we investigated the effect of RE in palmitate-induced resistant L6 myotubes. Exposure of myotubes to palmitate reduced the -stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the -stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the -stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell and further studies are required to explore its antidiabetic properties.

Keyword: insulin resistance

Alternation of plasma fatty acids composition and desaturase activities in children with liver steatosis.

The aim of this study was to investigate changes in plasma fatty acids proportions and estimated desaturase activities for variable grading of liver steatosis in children.In total, 111 schoolchildren (aged 8-18 years) were included in the analysis from March 2015 to August 2016. Anthropometric evaluation, liver ultrasound examination and scoring for nonalcoholic fatty liver disease (NAFLD score = 0-6), and biochemical and plasma fatty acids analysis were performed. We compared the composition ratio of fatty acids between children with high-grade liver steatosis (NAFLD score = 4-6), low-grade liver steatosis (NAFLD score = 1-3), and healthy controls (NAFLD score = 0). In addition, correlation coefficients (r) between NAFLD score, metabolic variables, and estimated activity of desaturase indices (stearoyl-coenzyme A desaturase-1 (SCD1), delta-5 and delta-6 desaturase) were calculated.Compared with healthy controls, children with liver steatosis showed a higher proportion of monounsaturated fatty acids (21.16 ± 2.81% vs. 19.68 ± 2.71%, p = 0.024). In addition, children with high- grade liver steatosis exhibited higher proportions of (C16:0), palmitoleic (C16:1n-7), dihomo-γ-linolenic (C20:3n-6), adrenic (C22:4n-6), and docosapentaenoic (C22:5n-6); and lower proportions of eicosapentaenoic (C20:5n-3) (P< 0.05). In all subjects, the NAFLD score was positively correlated with body mass index (BMI) (kg/m2) (r = 0.696), homeostasis model of assessment ratio-index (HOMA-IR) (r = 0.510), SCD1(16) (r = 0.273), and the delta-6 index (r = 0.494); and inversely associated with the delta-5 index (r = -0.443).Our current data suggested that children with liver steatosis was highly associated with obesity, and . In addition, increased endogenous lipogenesis through altered desaturase activity may contribute to the progression of liver steatosis in children.

Keyword: insulin resistance

Effect of weight loss on circulating fatty profiles in overweight subjects with high visceral fat area: a 12-week randomized controlled trial.

Significant associations between visceral fat and alterations in plasma fatty acids have been identified in overweight individuals. However, there are scant data regarding the relationships of the visceral fat area (VFA) with the plasma fatty profiles and desaturase activities following weight loss. We investigated the effect of weight loss with mild calorie restriction on the circulating fatty profiles and desaturase activities in nondiabetic overweight subjects with high VFA.Eighty overweight subjects with high VFA (L4 VFA ≥100\xa0cm) were randomized into the 12-week mild-calorie-restriction (300\xa0kcal/day) or control groups.Comparison of the percent of body weight changes between groups revealed that the weight-loss group had greater reductions in body weight. The VFA decreased by 17.7\xa0cm from baseline in the weight-loss group\xa0(P < 0.001). At follow-up, the weight-loss group showed greater reductions in serum triglycerides, , and HOMA-IR than the control group. Significantly greater reductions in total saturated fatty acids, , stearic , total monounsaturated fatty acids, palmitoleic , oleic , eicosadienoic , and dihomo-γ-linolenic levels were detected in the weight-loss group compared with the control group after adjusting for baseline values. Following weight loss, C16 Δ9-desaturase activity was significantly decreased and Δ5-desaturase activity was significantly increased, and the changes were greater in the weight-loss group than in the control group.The results suggest that mild weight loss improves abdominal obesity, overall fatty profiles, and desaturase activities; therefore, mild calorie restriction has potential health benefits related to obesity-related diseases in overweight subjects with high VFA.. Retrospectively registered 11 December 2016.

Keyword: insulin resistance

Kukoamine A attenuates and fatty liver through downregulation of Srebp-1c.

Nonalcoholic fatty liver disease (NAFLD) refers to a pathological condition of hepatic steatosis. is believed to be the key mechanism mediating initial accumulation of fat in the liver, resulting in hepatic steatosis. Kukoamine A (KuA), a spermine alkaloid, is a major bioactive component extracted from the root barks of Lycium chinense (L. chinense) Miller. In the current study, we aimed to explore the possible effect of KuA on and fatty liver. We showed that KuA significantly inhibited the increase of fasting blood glucose level and level, and the glucose levels in response to glucose and load in HFD-fed mice, which was in a dose-dependent manner. KuA dose-dependently decreased the histological injury of liver, levels of hepatic triglyceride (TG), and serum AST and ALT activities in HFD-fed mice. The increase of serum levels of TNFɑ, IL-1β, IL-6 and C reactive protein in HFD-fed mice was inhibited by KuA. HFD feeding-induced increase of hepatic expression of Srebp-1c and its target genes, including fatty synthase (FAS) and acetyl CoA carboxylase 1 (ACC1), was significantly inhibited by KuA. Moreover, upregulation of Srebp-1c notably inhibited KuA-induced improvement of -stimulated glucose uptake, decrease of lipid accumulation and HO level in -treated AML-12 cells. In conclusion, we reported that KuA inhibited Srebp-1c and downstream genes expression and resulted in inhibition of lipid accumulation, inflammation, and oxidative stress. Overall, our results provide a better understanding of the pharmacological activities of KuA against and hepatic steatosis.Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Keyword: insulin resistance

Ceramide stearic to ratio predicts incident diabetes.

Ceramide lipids have a role in the development of , diabetes and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident diabetes.A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the metabolic syndrome (Prevent Metabolic Syndrome [PrevMetSyn], n\u2009=\u2009371). Diabetes risk score models were developed to estimate the 10\xa0year risk of incident diabetes.Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident diabetes (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and diabetes risk scores significantly decreased in individuals who had 5% or more weight loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident diabetes, and may be modulated by lifestyle intervention.

Keyword: insulin resistance

Alterations in branched-chain amino kinetics in nonobese but -resistant Asian men.

Branched-chain amino acids (BCAAs) are elevated in the -resistant (IR) state. The reasons for this increase remain unclear, but it may be related to abnormalities in BCAA metabolism and free fatty (FFA) metabolism.In this study, we quantified BCAA and FFA kinetics of IR and -sensitive (IS) nonobese Asian men with the use of stable-isotope tracers. We hypothesized that in addition to greater substrate flux, the BCAA oxidative pathway is also impaired to account for the higher plasma BCAA concentration in the IR state.We recruited 12 IR and 14 IS nonobese and healthy Asian men. Oral-glucose-tolerance tests (OGTTs) were performed to quantify sensitivity, and subjects underwent 2 stable-isotope infusion studies. [U-13C6]Leucine was infused to measure leucine flux and oxidation as indexes of BCAA metabolism, whereas [U-13C16]palmitate was infused to measure palmitate flux and oxidation to represent FFA metabolism, The 2H2O dilution method was used to estimate body composition.IR subjects had greater adiposity and significantly higher fasting and post-OGTT glucose and concentrations compared with the IS group. However, none of the subjects were diabetic. Despite similar dietary protein intake, IR subjects had a significantly higher plasma BCAA concentration and greater leucine flux. Leucine oxidation was also greater in the IR group, but the relation between leucine oxidation and flux was significantly weaker in the IR group than in the IS group (r\xa0=\xa00.530 compared with\xa00.695, P\xa0<\xa00.0388 for differences between slope). FFA oxidation was, however, unaffected despite higher FFA flux in the IR group.The higher plasma BCAA concentration in healthy nonobese individuals with IR is associated with a weaker relation between BCAA oxidation and BCAA flux and this occurs in the presence of accelerated FFA flux and oxidation.

Keyword: insulin resistance

Histone methyltransferase G9a modulates hepatic signaling via regulating HMGA1.

Hepatic sensitivity is critical for glucose homeostasis, and is a fundamental syndrome found in various metabolic disorders, including obesity and type 2 diabetes. Despite considerable studies on the mechanisms of hepatic , the link between epigenetic regulation and the development of remains elusive. Here, we reported that G9a/EHMT2, a histone methyltransferase, was markedly decreased in the liver of db/db mice and high-fat diet (HFD)-fed mice. In cultured hepatic cells, G9a knockdown resulted in downregulation of receptor, p-AKT and p-GSK3β; while upon upregulation, G9a prevented the - or glucosamine-induced by preserving the normal level of receptor and integrity of signaling. Further mechanistic study suggested that G9a regulated the expression level of high mobility group AT-hook 1 (HMGA1), a key regulator responsible for the transcription of receptor (INSR) gene. Overexpression of HMGA1 normalized the impaired signaling in G9a knockdown hepatic cells. Importantly, in db/db mice, restoring the expression level of G9a not only upregulated HMGA1 level and improved the impaired hepatic signaling, but also alleviated hyperglycemia and hyperinsulinemia. Together, our results revealed a novel role for G9a in modulating signaling, at least in part, depending on its regulatory function on HMGA1.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Acute and Repeated Treatment with 5-PAHSA or 9-PAHSA Isomers Does Not Improve Glucose Control in Mice.

Fatty esters of hydroxylated fatty acids (FAHFAs) were discovered as a novel class of endogenous mammalian lipids whose profound effects on metabolism have been shown. In the current study, in\xa0vitro and in\xa0vivo the metabolic effects of two of these FAHFAs, namely -5- (or -9) -hydroxy-stearic (5- or 9-PAHSA, respectively) were profiled. In DIO mice fed with differentially composed low- or high-fat diets, acute and subchronic treatment with 5-PAHSA and 9-PAHSA alone, or in combination, did not significantly improve the deranged metabolic status. Neither racemic 5- or 9-PAHSA, nor the enantiomers were able to: (1) increase basal or -stimulated glucose uptake in\xa0vitro, (2) stimulate GLP-1 release from GLUTag cells, or (3) induce GSIS in rat, mouse, or human islets or in a human pancreatic β cell line. Therefore, our data do not support the\xa0further development of PAHSAs or their derivatives for the control of and hyperglycemia.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Inhibition of protein kinase R protects against -induced inflammation, oxidative stress, and apoptosis through the JNK/NF-kB/NLRP3 pathway in cultured H9C2 cardiomyocytes.

Double-stranded RNA-dependent protein kinase (PKR) is a critical regulator of apoptosis, oxidative stress, and inflammation under hyperlipidemic and conditions. Saturated free fatty acids, such as (PA), are known inducers of apoptosis in numerous cell types. However, the underlying molecular mechanism is not fully understood. The aim of the present study was to examine the effect of PA on cultured rat H9C2 cardiac myocytes cells and to investigate the PKR mediated harmful effects of PA in vitro in cultured cardiomyocytes.PKR expression was determined by immunofluorescence and immunoblotting. Oxidative stress and apoptosis were determined by flow cytometry and assay kits. The expression of different gene markers of apoptosis, oxidative stress, and inflammation were measured by Western blot analysis and reverse transcription polymerase chain reaction.PKR expression, reactive oxygen species levels as well as apoptosis were increased in PA-treated cultured H9C2 cardiomyocytes. The harmful effects of PA were attenuated by a selective PKR inhibitor, C16. Moreover, we observed that upregulation of c-Jun N-terminal kinase (JNK), nuclear factor-kB (NF-kB) and NACHT, LRR and PYD domains-containing protein 3 (NLRP3) pathways is associated with increased expression of interleukin 6 and tumor necrosis factor-α in PA-treated cardiomyocytes and attenuation by a selective PKR inhibitor.Our study reports, for the first time, that PKR-mediated harmful effects of PA in cultured cardiomyocytes via activation of JNK, NF-kB, and NLRP3 pathways. Inhibition of PKR is one of the possible mechanistic approaches to inhibit inflammation, oxidative stress, and apoptosis in lipotoxicity-induced cardiomyocyte damage.© 2018 Wiley Periodicals, Inc.

Keyword: insulin resistance

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores sensitivity in experimental models.

Type 2 diabetes mellitus (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of . Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring sensitivity and glycaemic control in models and revealing its underlying mechanism. (PA)-treated LO2 cells and high fat diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, fatty acids and cholesterol without affecting the serum levels. Moreover, our data demonstrated that YMS could restore glucose and lipid homeostasis via facilitating transportation and reactivating PI3K/Akt signaling in both PA-treated cells and liver, gastrocnemius and brown fat of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with , particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: insulin resistance

is an intracellular signaling molecule involved in disease development.

Emerging evidence shows that (PA), a common fatty in the human diet, serves as a signaling molecule regulating the progression and development of many diseases at the molecular level. In this review, we focus on its regulatory roles in the development of five pathological conditions, namely, metabolic syndrome, cardiovascular diseases, cancer, neurodegenerative diseases, and inflammation. We summarize the clinical and epidemiological studies; and also the mechanistic studies which have identified the molecular targets for PA in these pathological conditions. Activation or inactivation of these molecular targets by PA controls disease development. Therefore, identifying the specific targets and signaling pathways that are regulated by PA can give us a better understanding of how these diseases develop for the design of effective targeted therapeutics.

Keyword: insulin resistance

Modulation of and the Adipocyte-Skeletal Muscle Cell Cross-Talk by LCn-3PUFA.

The cross-talk between skeletal muscle and adipose tissue is involved in the development of (IR) in skeletal muscle, leading to the decrease in the anabolic effect of . We investigated if the long chain polyunsaturated n-3 fatty acids (LCn-3PUFA), eicosapentaenoic and docosapentaenoic acids (EPA and DPA, respectively) could (1) regulate the development of IR in 3T3-L1 adipocytes and C2C12 muscle cells and (2) inhibit IR in muscle cells exposed to conditioned media (CM) from -resistant adipocytes. Chronic (CI) treatment of adipocytes and (PAL) exposure of myotubes were used to induce IR in the presence, or not, of LCn-3PUFA. EPA (50 µM) and DPA (10 µM) improved PAL-induced IR in myotubes, but had only a partial effect in adipocytes. CM from adipocytes exposed to CI induced IR in C2C12 myotubes. Although DPA increased the mRNA levels of genes involved in fatty (FA) beta-oxidation and signaling in adipocytes, it was not sufficient to reduce the secretion of inflammatory cytokines and prevent the induction of IR in myotubes exposed to adipocyte\'s CM. Treatment with DPA was able to increase the release of adiponectin by adipocytes into CM. In conclusion, DPA is able to protect myotubes from PAL-induced IR, but not from IR induced by CM from adipocytes.

Keyword: insulin resistance

Effects of high-fat diet and AMP-activated protein kinase modulation on the regulation of whole-body lipid metabolism.

Metabolic flexibility, the capacity to adapt to fuel availability for energy production, is crucial for maintaining whole-body energy homeostasis. An inability to adequately promote FA utilization is associated with lipid accumulation in peripheral tissues and contributes to the development of . In vivo assays to quantify whole-body lipid oxidation in mouse models of are lacking. We describe a method for assessing whole-body FA oxidation in vivo, as well as tissue-specific lipid uptake in conscious mice. The method relies on intravenous administration of [9,10-H(N)] combined with a non-β-oxidizable palmitate analog, [1-C]2-bromopalmitic . Pretreatment with etomoxir, a CPT1 inhibitor that prevents the shuttling of FAs into mitochondria, markedly reduced the appearance of the β-oxidation product HO in circulation and reduced lipid uptake by oxidative tissues including heart and soleus muscle. Whole-body fatty oxidation was unaltered between chow- or high-fat-fed WT and transgenic mice expressing a mutant form of the AMPK γ3 subunit (AMPKγ3) in skeletal muscle. High-fat feeding increased lipid oxidation in WT and AMPKγ3 transgenic mice. In conclusion, this technique allows for the assessment of the effect of pharmaceutical agents, as well as gene mutations, on whole-body FA oxidation in mice.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: insulin resistance

Hyperlipidemia-induced hepassocin in the liver contributes to in skeletal muscle.

Hepassocin (HPS) has recently been identified as a novel hepatokine that causes hepatic steatosis. However, the role of HPS in the development of in skeletal muscle under obesity remains unclear. The effect of hyperlipidemia on hepatic HPS expression was evaluated in primary hepatocytes and liver of mice. HPS-mediated signal pathways were explored using small interfering (si) RNAs of specific genes or inhibitors. We found that treatment of primary hepatocytes with palmitate could induce HPS expression through C/EBPβ-mediated transcriptional activation. Furthermore, increased HPS expression was observed in the liver of high fat diet (HFD)-fed or tunicamycin-treated mice. Pretreatment with 4-phenylbutyrate (4-BPA) (an endoplasmic reticulum (ER) stress inhibitor) and suppression of p38 by siRNA abrogated the effect of palmitate on HPS expression in primary hepatocytes. Treatment of differentiated C2C12\xa0cells with recombinant HPS caused c-Jun N-terminal kinase (JNK) phosphorylation and impairment of sensitivity in a dose-dependent manner. siRNA-mediated suppression of JNK reduced the effect of HPS on signaling. Furthermore, the suppression of epidermal growth factor receptor (EGFR) by siRNA mitigated both HPS-induced JNK phosphorylation and . In addition, HPS did not affect inflammation and ER stress in differentiated C2C12\xa0cells. In conclusion, we elucidated that ER stress induced by palmitate could increase the expression of HPS in hepatocytes and further contribute to the development of in skeletal muscle via EGFR/JNK-mediated pathway. Taken together, we suggest that HPS could be a therapeutic target for obesity-linked .Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

BPN, a marine-derived PTP1B inhibitor, activates signaling and improves in C2C12 myotubes.

is a key feature of type 2 diabetes mellitus (T2DM) and is characterized by defects in signaling. Protein tyrosine phosphatase 1B (PTP1B) is a major negative regulator of signaling cascade and has attracted intensive investigation in recent T2DM therapy study. BPN, a marine-derived bromophenol compound, was isolated from the red alga Rhodomela confervoides. This study investigated the effects of BPN on the signaling pathway in -resistant C2C12 myotubes by inhibiting PTP1B. Molecular docking study and analysis of small- molecule interaction with PTP1B all showed BPN inhibited PTP1B activity via binding to the catalytic site through hydrogen bonds. We then found that BPN permeated into C2C12 myotubes, on the one hand, activated signaling in an -independent manner in C2C12 cells; on the other hand, ameliorated palmitate-induced through augmenting sensitivity. Moreover, our studies also showed that PTP1B inhibition by BPN increased glucose uptake in normal and -resistant C2C12 myotubes through glucose transporter 4 (GLUT4) translocation. Taken together, BPN activates signaling and alleviates and represents a potential candidate for further development as an antidiabetic agent.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Nutritional modulation of metabolic inflammation.

Metabolic inflammation is a very topical area of research, wherein aberrations in metabolic and inflammatory pathways probably contribute to atherosclerosis, (IR) and type 2 diabetes. Metabolic insults arising from obesity promote inflammation, which in turn impedes signalling and reverse cholesterol transport (RCT). Key cells in the process are metabolically activated macrophages, which up-regulate both pro- and anti-inflammatory pathways in response to lipid spillover from adipocytes. Peroxisome proliferator-activated receptors and AMP-activated protein kinase (AMPK) are regulators of cellular homeostasis that influence both inflammatory and metabolic pathways. Dietary fats, such as saturated fatty acids (SFAs), can differentially modulate metabolic inflammation. , in particular, is a well-characterized nutrient that promotes metabolic inflammation via the NLRP3 (the nod-like receptor containing a pyrin domain) inflammasome, which is partly attributable to AMPK inhibition. Conversely, some unsaturated fatty acids are less potent agonists of metabolic inflammation. For example, monounsaturated fatty does not reduce AMPK as potently as SFA and n-3 polyunsaturated fatty acids actively resolve inflammation via resolvins and protectins. Nevertheless, the full extent to which nutritional state modulates metabolic inflammation requires greater clarification.© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: insulin resistance

Hepatic transcriptome analysis from HFD-fed mice defines a long noncoding RNA regulating cellular cholesterol levels.

To elucidate the transcriptomic changes of long noncoding RNAs (lncRNAs) in high-fat diet (HFD)-fed mice, we defined their hepatic transcriptome by RNA sequencing. Aberrant expression of 37 representative lncRNAs and 254 protein-coding RNAs was observed in the livers of HFD-fed mice with compared with the livers from control mice. Of these, 24 lncRNAs and 179 protein-coding RNAs were upregulated, whereas 13 lncRNAs and 75 protein-coding RNAs were downregulated. Functional analyses showed that the aberrantly expressed protein-coding RNAs were enriched in various lipid metabolic processes and in the signaling pathway. Genomic juxtaposition and coexpression patterns identified six pairs of aberrantly expressed lncRNAs and protein-coding genes, consisting of five lncRNAs and five protein-coding genes. Four of these protein-coding genes are targeted genes upregulated by PPARα. As expected, the corresponding lncRNAs were significantly elevated in AML12 cells treated with or the PPARα agonist, WY14643. In Hepa1-6 cells, knockdown of NONMMUG027912 increased the cellular cholesterol level, the expression of cholesterol biosynthesis genes and proteins, and the HMG-CoA reductase activity. This genome-wide profiling of lncRNAs in HFD-fed mice reveals one lncRNA, NONMMUG027912, which is potentially regulated by PPARα and is implicated in the process of cholesterol biosynthesis.Copyright © 2019 Chen et al. Published by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: insulin resistance

Free Fatty Acids Induce Autophagy and LOX-1 Upregulation in Cultured Aortic Vascular Smooth Muscle Cells.

Elevation of free fatty acids (FFAs) is known to affect microvascular function and contribute to obesity-associated , hypertension, and microangiopathy. Proliferative and synthetic vascular smooth muscle cells (VSMCs) increase intimal thickness and destabilize atheromatous plaques. This study aimed to investigate whether saturated (PA) and monounsaturated oleic (OA) modulate autophagy activity, cell proliferation, and vascular tissue remodeling in an aortic VSMC cell line. Exposure to PA and OA suppressed growth of VSMCs without apoptotic induction, but enhanced autophagy flux with elevation of Beclin-1, Atg5, and LC3I/II. Cotreatment with autophagy inhibitors potentiated the FFA-suppressed VSMC growth and showed differential actions of PA and OA in autophagy flux retardation. Both FFAs upregulated lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) but only OA increased LDL uptake by VSMCs. Mechanistically, FFAs induced hyperphosphorylation of Akt, ERK1/2, JNK1/2, and p38 MAPK. All pathways, except OA-activated PI3K/Akt cascade, were involved in the LOX-1 upregulation, whereas blockade of PI3K/Akt and MEK/ERK cascades ameliorated the FFA-induced growth suppression on VSMCs. Moreover, both FFAs exhibited tissue remodeling effect through increasing MMP-2 and MMP-9 expression and their gelatinolytic activities, whereas high-dose OA significantly suppressed collagen type I expression. Conversely, siRNA-mediated LOX-1 knockdown significantly attenuated the OA-induced tissue remodeling effects in VSMCs. In conclusion, OA and PA enhance autophagy flux, suppress aortic VSMC proliferation, and exhibit vascular remodeling effect, thereby leading to the loss of VSMCs and interstitial ECM in vascular walls and eventually the instability of atheromatous plaques. J. Cell. Biochem. 118: 1249-1261, 2017.© 2016 Wiley Periodicals, Inc.

Keyword: insulin resistance

Withdrawal from high-carbohydrate, high-saturated-fat diet changes saturated fat distribution and improves hepatic low-density-lipoprotein receptor expression to ameliorate metabolic syndrome in rats.

The "lipid hypothesis" determined that saturated fatty (SFA) raises low-density lipoprotein cholesterol, thereby increasing the risk for metabolic syndrome (MetS). The aim of this study was to investigate the effect of subchronic withdrawal from a high-carbohydrate, high-saturated fat (HCHF) diet during MetS with reference to changes in deleterious SFA (C12:0, lauric ; C14:0, myristic ; C16:0, ; C18:0, stearic ) distribution in liver, white adipose tissue (WAT), and feces.MetS induced by prolonged feeding of an HCHF diet in Wistar albino rat is used as a model of human MetS. The MetS-induced rats were withdrawn from the HCHF diet and changed to a basal diet for final 4\xa0wk of the total experimental duration of 16\xa0wk. SFA distribution in target tissues and hepatic low-density lipoprotein receptor (LDLr) expression were analyzed.Analyses of changes in SFA concentration of target tissues indicate that C16:0 and C18:0 reduced in WAT and liver after withdrawal of the HCHF diet. There was a significant (P\xa0<\xa00.001) decrease in fecal C12:0 with HCHF feeding, which significantly (P\xa0<\xa00.01) increased after withdrawal of this diet. Also, an improvement in expression of hepatic LDLr was observed after withdrawal of HCHF diet.The prolonged consumption of an HCHF diet leads to increased SFA accumulation in liver and WAT, decreased SFA excretion, and reduced hepatic LDLr expression during MetS, which is prominently reversed after subchronic withdrawal of the HCHF diet. This can contribute to better understanding of the metabolic fate of dietary SFA during MetS and may apply to the potential reversal of complications by the simple approach of nutritional modification.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Fatty composition in serum cholesterol esters and phospholipids is linked to visceral and subcutaneous adipose tissue content in elderly individuals: a cross-sectional study.

Visceral adipose tissue (VAT) and truncal fat predict cardiometabolic disease. Intervention trials suggest that saturated fatty acids (SFA), e.g. , promote abdominal and liver fat storage whereas polyunsaturated fatty acids (PUFA), e.g. linoleic , prevent fat accumulation. Such findings require investigation in population-based studies of older individuals. We aimed to investigate the relationships of serum biomarkers of PUFA intake as well as serum levels of , with abdominal and total adipose tissue content.In a population-based sample of 287 elderly subjects in the PIVUS cohort, we assessed fatty composition in serum cholesterol esters (CE) and phospholipids (PL) by gas chromatography and the amount of VAT and abdominal subcutaneous (SAT) adipose tissue by magnetic resonance imaging (MRI), liver fat by MR spectroscopy (MRS), and total body fat, trunk fat and leg fat by dual-energy X-ray absorptiometry (DXA). was estimated by HOMA-IR.VAT and trunk fat showed the strongest correlation with (r\u2009=\u20090.49, P\u2009<\u20090.001). Linoleic in both CE and PL was inversely related to all body fat depots (r\u2009=\u2009-0.24 to -0.33, P\u2009<\u20090.001) including liver fat measured in a sub-group (r\u2009=\u2009-0.26, P\u2009<\u20090.05, n\u2009=\u200973), whereas n-3 PUFA showed weak inverse (18:3n-3) or positive (20:5n-3) associations. in CE, but not in PL, was directly correlated with VAT (r\u2009=\u20090.19, P\u2009<\u20090.001) and trunk fat (r\u2009=\u20090.18, P\u2009=\u20090.003). Overall, the significant associations remained after adjusting for energy intake, height, alcohol, sex, smoking, education and physical activity. The inverse correlation between linoleic and VAT remained significant after further adjustment for total body fat.Serum linoleic is inversely related to body fat storage including VAT and trunk fat whereas was less consistently but directly associated, in line with recent feeding studies. Considering the close link between VAT and , a potential preventive role of plant-based PUFA in VAT accumulation warrants further study.

Keyword: insulin resistance

Human relaxin-2 attenuates hepatic steatosis and fibrosis in mice with non-alcoholic fatty liver disease.

Human relaxin-2 reduces hepatic fibrosis in mice. However, the effects of relaxin-2 on hepatic steatosis and fibrosis in animals with non-alcoholic fatty liver disease (NAFLD) remain to be elucidated. C57BL/6 mice fed a high-fat diet (HFD) or methionine-choline-deficient (MCD) diet were randomly assigned to receive recombinant human relaxin-2 (25 or 75\u2009μg/kg/day) or vehicle for 4 weeks. In HFD-fed mice, relaxin-2 decreased systemic and reduced body weight, epididymal fat mass and serum leptin and concentrations. In livers of HFD-fed mice, relaxin-2 attenuated steatosis and increased phosphorylation of receptor substrate-1, Akt and endothelial nitric oxide synthase (eNOS), and activated genes that regulate fatty oxidation and suppressed acetyl-CoA carboxylase. Relaxin-2 had no direct anti-steatotic effect on primary mouse hepatocytes, but S-nitroso-N-acetylpenicillamine attenuated -induced steatosis and activated genes regulating fatty oxidation in hepatocytes. In mice fed an MCD diet, relaxin-2 attenuated steatosis, inflammation and fibrosis. Relaxin-2 increased eNOS and Akt phosphorylation and transcript levels of cytochrome P450-4a10 and decreased acetyl-CoA carboxylase in MCD-fed mouse livers. Moreover, expression levels of Kupffer cell activation, hepatic stellate cell activation and hepatocyte apoptosis were decreased in MCD diet-fed mice receiving relaxin-2. In conclusion, relaxin-2 reduces hepatic steatosis by activating intrahepatic eNOS in HFD-fed mice and further attenuates liver fibrosis in MCD diet-fed mice. Therefore, human relaxin-2 is a potential therapeutic treatment for NAFLD.

Keyword: insulin resistance

TNF-α stimulates endothelial transcytosis and promotes .

Persistent elevation of plasma TNF-α is a marker of low grade systemic inflammation. (PA) is the most abundant type of saturated fatty in human body. PA is bound with albumin in plasma and could not pass through endothelial barrier freely. Albumin-bound PA has to be transported across monolayer endothelial cells through intracellular transcytosis, but not intercellular diffusion. In the present study, we discovered that TNF-α might stimulate PA transcytosis across cardiac microvascular endothelial cells, which further impaired the -stimulated glucose uptake by cardiomyocytes and promoted . In this process, TNF-α-stimulated endothelial autophagy and NF-κB signaling crosstalk with each other and orchestrate the whole event, ultimately result in increased expression of fatty transporter protein 4 (FATP4) in endothelial cells and mediate the increased PA transcytosis across microvascular endothelial cells. Hopefully the present study discovered a novel missing link between low grade systemic inflammation and .

Keyword: insulin resistance

Increased serum concentration of ceramides in obese children with nonalcoholic fatty liver disease.

Hepatic lipid accumulation is closely related to the development of , which is regarded as one of the most significant risk factors of nonalcoholic fatty liver disease (NAFLD). Although the exact molecular pathway leading to impaired signaling has not been definitively established, ceramides are suspected mediators of lipid induced hepatic . Therefore, the aim of the study was to evaluate the serum ceramides concentration in obese children with NAFLD.The prospective study included 80 obese children (aged 7-17\xa0years, median 12\xa0years) admitted to our Department to diagnose initially suspected liver disease. Patients with viral hepatitis (HCV, HBV, CMV), autoimmune (AIH), toxic and metabolic (Wilson\'s disease, alfa-1-antitrypsin deficiency) liver diseases and celiac disease were excluded. NAFLD was diagnosed based on pediatric diagnostic criteria in obese children with liver steatosis in ultrasound (US) as well as elevated alanine transaminase (ALT) serum activity after exclusion of other major liver diseases listed before. Ultrasonography was used as a screening method and for qualitative assessment of the steatosis degree (graded according to Saverymuttu scale). Advanced steatosis was defined as a score\u2009>\u20091. The total intrahepatic lipid content (TILC) was assessed by magnetic resonance proton spectroscopy (HMRS) which is the most accurate technique for assessment of ectopic fat accumulation. Fasting serum concentration of ceramides was measured in 62 children.NAFLD was diagnosed in 31 children. Significant, positive correlation was found between total serum concentration of ceramides and (r\xa0=\u20090.3, p\xa0=\u20090.02) and HOMA-IR (r\xa0=\u20090.28, p\xa0=\u20090.03). Total ceramide concentration as well as specific fatty -ceramides (FA-ceramides) concentrations, namely: myristic, , palmitoleic, stearic, oleic, behenic and lignoceric were significantly higher (p\xa0=\u20090.004, p\xa0=\u20090.003, p\xa0=\u20090.007, p\xa0<\u20090.001, p\xa0=\u20090.035, p\xa0=\u20090.008, p\xa0=\u20090.003, p\xa0=\u20090.006, respectively) in children with NAFLD compared to controls (n\xa0=\u200914). Moreover, children with NAFLD had significantly higher activity of ALT (p\xa0<\u20090.001) and GGT (p\xa0<\u20090.001), HOMA-IR (p\xa0=\u20090.04), BMI (p\xa0=\u20090.046), waist circumference (p\xa0=\u20090.01) steatosis grade in ultrasound (p\xa0<\u20090.001) and TILC in HMRS (p\xa0<\u20090.001) compared to children without NAFLD. We did not find significant differences in total and FA-ceramide species concentrations between children with mild (grade 1) and advanced liver steatosis in ultrasonography (grade 2-3).Elevated ceramide concentrations in obese patients together with their significant correlation with parameters suggest their association with molecular pathways involved in signaling impairment known to be strongly linked to pathogenesis of non-alcoholic fatty liver disease.

Keyword: insulin resistance

Melatonin improves and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with and type 2 diabetes. The aim of this study was to determine whether melatonin improves hepatic and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve in FETUA-overexpressing cells, indicating that improvement in by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced body weight gain to one-third of that of the HFD group and hepatic steatosis. sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the liver and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: insulin resistance

The BACE1 product sAPPβ induces ER stress and inflammation and impairs signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in -independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and fatty oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, , PGC-1α down-regulation, and fatty oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and in skeletal muscle and gluconeogenesis in liver.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Vasoreactivity of isolated aortic rings from dyslipidemic and resistant inducible nitric oxide synthase knockout mice.

Recent study from this lab indicated enhanced susceptibility of iNOS KO mice for diet induced obesity (DIO) and systemic (IR) as compared to C57BL/6 (WT) mice. The present study investigates aortic vasoreactivity in high fat diet (HFD) induced resistant iNOS KO mice. WT and iNOS KO mice were fed with 45% HFD/10% LFD for ten weeks. Systemic IR was assessed via measurement of circulating lipids, glucose, and ; while phenylephrine (PE)/acetylcholine (ACh) induced responses were monitored in the isolated aortic rings. To understand the mechanism, qPCR or Western blotting experiments were performed in aorta and Ea.hy926\u202fcells. After 10 weeks of HFD feeding, significant increase in the body weight/fat mass, augmented circulating lipids, glucose, and inflammatory cytokines along with impaired acetylcholine induced aortic vasorelaxation and enhanced iNOS expression was observed in the aortic tissue of WT mice. In the aminoguanidine (AG, 20\u202fmg/kg for 4 weeks) treated WT mice and also in iNOS KO mice, acetylcholine induced vasorelaxation was significantly preserved. Further, acetylcholine mediated vasorelaxation correlated with increased eNOS phosphorylation at Ser1177 residue in the iNOS KO mice and same was also observed in the iNOS silenced Ea.hy926\u202fcells. Moreover, treatment of Ea.hy926\u202fcells with or TNFα also caused a significant decrease in eNOS activity, which was reversed in iNOS silenced Ea.hy926\u202fcells suggesting the role of iNOS in the reduction of eNOS activity. The study thus implies a critical role of iNOS in vascular diseases associated with dyslipidemia/IR.Copyright © 2019. Published by Elsevier B.V.

Keyword: insulin resistance

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, , and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: insulin resistance

Hepatocyte growth factor alleviates hepatic and lipid accumulation in high-fat diet-fed mice.

Type 2 diabetes mellitus is frequently accompanied by fatty liver disease. Lipid accumulation within the liver is considered as one of the risk factors for . Hepatocyte growth factor (HGF) is used to treat liver dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting and homeostatic model assessment of were calculated for sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/fatty synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: insulin resistance

[The disturbance of unification of coupled biochemical reactions in synthesis of endogenous ω-9 oleic . The to , stearic triglycerides and pathogenesis of eruptive xanthomata].

The eruptive xanthomata are formed in vivo under realization of biological function of endoecology. The xanthomata are formed in tissues by early in phylogenesis resident macrophages at absorption of secreted by hepatocytes aphysiological stearic lipoproteins of very low density with high content of the same name triglycerides down to tristearate. In these lipoproteins of very low density, by force of aphysiologically high hydrophobicity, stearic triglycerides are not hydrolyzed by post-heparin lipoproteinlipase. They both do not associate apoE and form apoE/B-J00 ligands. The formation of stearic lipoproteins of very low density occurred at impairment of function of coupled biochemical reactions in synthesis of physiological ω-9 oleic mono unsaturated fatty in hepatocytes. To synthesize endogenous oleic mono unsaturated fatty the late in phylogenesis expresses two enzymes of coupled biochemical reactions: palmitoyl-KoA-elongase andstearyl-KoA-desaturase, activating synthesis of fatty acids following the path glucose-endogenous unsaturated fatty -stearic unsaturated fatty -oleic mono unsaturated fatty . The uncoupling of enzymes of coupling synthesis forms in hepatocytes surplus of stearic mono unsaturated fatty , stearic triglycerides and of the same name aphysiologic lipoproteins of very low density. During inhibition of the second enzyme the first one continues to actively produce stearic unsaturated fatty which the second enzyme, already uncoupled, does not convert into oleic unsaturated fatty . By absorbing aphysiologic ligand-free stearic lipoproteins of very low density in biologic reaction of endoecology, phylogenetically early macrophages convert into foam cells initiating aphysiologic biological reaction of transcytosis, biologic reaction of inflammation, biologic reaction of apoptosis and formation of eruptive xanthomata. The lipids of eruptive xanthomata: such endogenous stearic triglycerides as tristearate, tripalmitate, exogenous carotenoids, phospholipids and unesterified cholesterol.

Keyword: insulin resistance

Resveratrol attenuates type 2 diabetes mellitus by mediating mitochondrial biogenesis and lipid metabolism via Sirtuin type 1.

The rising incidence of type 2 diabetes mellitus (T2DM) is a major public health problem and novel therapeutic strategies are required to prevent and treat T2DM. It has been demonstrated that resveratrol (RSV) may prevent T2DM by targeting Sirtuin type 1 (SIRT1), indicating that SIRT1 may be a novel therapeutic target for T2DM prevention. In the present study, a T2DM rat model was established by administering a high fat diet and streptozotocin (STZ) injections. Measurements of blood glucose and confirmed successful establishment of the T2DM model. RSV was used to treat rats with STZ-induced T2DM and the results indicated that RSV reversed the STZ-induced downregulation of peroxisome proliferator-activated receptor-γ coactivator-1α, SIRT1 and forkhead box protein O 3a. Furthermore, RSV modulated the activity of superoxide dismutase and malondialdehyde, which are associated with oxidative stress. , cells from the insulinoma cell line clone 1E were pretreated with (PA) to simulate a high fat environment. The results of reverse transcription-quantitative polymerase chain reaction indicated that PA suppressed the expression of SIRT1 in a dose- and time-dependent manner. Furthermore, PA modulated the expression of mitochondrial biogenesis-associated, lipid metabolism-associated and β-cell-associated genes, whereas RSV treatment ameliorated the PA-induced changes in the expression of these genes via SIRT1. The results of the present study suggest that RSV participates in the prevention of T2DM by regulating the expression of mitochondrial genes associated with biogenesis, lipid metabolism and β-cells via SIRT1. The results of the current study provide an insight into the mechanisms by which SIRT1 inhibits T2DM and may be used as a basis for future studies.

Keyword: insulin resistance

Association of dietary factors with severity of coronary artery disease.

Cardiovascular disease has taken epidemic proportions during past decades. Cardiovascular risk factors contribute to progression of coronary lesions, worsening the patient\'s prognosis. This study was planned to analyze the association of dietary factors with severity of coronary artery disease (CAD) in Indian patients.Three hundred patients with known coronary disease above the age of 25 years were included in this study. Blood samples were collected for biochemical markers. Patients were stratified according to severity of CAD [number of vessel involved-single (SVD), double (DVD), triple (TVD)].Mean age of the patient was 60.9\xa0±\xa012.4 years. Subjects with TVD, DVD, SVD in the study were 52.3%, 25.3% and 22.3% respectively. Patients with TVD had higher body mass index, triglycerides, HOMA-, hsCRP and lower high density cholesterol. Diabetes mellitus, hypertension and dyslipidemia were more common in TVD patients. Among macronutrients, patients with TVD had higher intake of carbohydrate and lower intake of protein and dietary fibers. There was no association of total fat intake with CAD, however, intake of was higher among patients with TVD. Intake of vitamins namely niacin, riboflavin, thiamine, B6, and vitamin-C decreased with increase in severity. With increase in severity of CAD, mineral intake (potassium, calcium, magnesium, phosphorus, sulfur, iron, chromium, copper, manganese, and zinc) decreased.Dietary factors are associated with severity of coronary artery disease. Low intake of protein, fiber, vitamins, minerals and high intake of carbohydrate and fat was associated with higher probability of having severe CAD.Copyright © 2016 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.

Keyword: insulin resistance

Influence of resveratrol on endoplasmic reticulum stress and expression of adipokines in adipose tissues/adipocytes induced by high-calorie diet or .

This study aimed to determine whether resveratrol treatment alleviates endoplasmic reticulum stress and changes the expression of adipokines in adipose tissues and cells.8-week-old male C57BL/6 mice were fed a high-calorie diet (HCD group) or high-calorie diet supplemented with resveratrol (high-calorie diet \u2009+\u2009resveratrol group) for 3 months. , serum lipids and proinflammatory indices, the size and inflammatory cell infiltration in subcutaneous and visceral adipose tissues were analyzed. The gene expressions of endoplasmic reticulum stress, adipokines, and inflammatory cytokines were determined. The induced mature 3T3-L1 cells were pretreated with resveratrol and then , and the gene expressions of endoplasmic reticulum stress, adipokines, and inflammatory cytokines were determined.Subcutaneous and visceral adipose tissues in the high-calorie diet-fed mice exhibited adipocyte hypertrophy, inflammatory activation, and endoplasmic reticulum stress. Resveratrol alleviated high-calorie diet-induced and endoplasmic reticulum stress, increased expression of SIRT1, and reversed expression of adipokines in varying degrees in both subcutaneous and visceral adipose tissues. The effects of resveratrol on -treated adipocytes were similar to those shown in the tissues.Resveratrol treatment obviously reversed adipocyte hypertrophy and by attenuating endoplasmic reticulum stress and inflammation, thus increasing the expression of SIRT1 and inverting the expression of adipokines in vivo and in vitro.

Keyword: insulin resistance

lncRNA MEG3 promotes hepatic by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑fat diet (HFD)‑fed mice to verify the role of MEG3 in hepatic in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 diabetes mellitus.

Keyword: insulin resistance

Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced in human skeletal muscle cells.

Free fatty is considered to be one of the major pathogenic factors of inducing . The association between iron disturbances and has recently begun to receive a lot of attention. Although skeletal muscles are a major tissue for iron utilization and storage, the role of iron in palmitate (PA)-induced is unknown. We investigated the molecular mechanism underlying iron dysregulation in PA-induced . Interestingly, we found that PA simultaneously increased intracellular iron and induced . The iron chelator deferoxamine dramatically inhibited PA-induced , and iron donors impaired sensitivity by activating JNK. PA up-regulated transferrin receptor 1 (tfR1), an iron uptake protein, which was modulated by iron-responsive element-binding proteins 2. Knockdown of tfR1 and iron-responsive element-binding proteins 2 prevented PA-induced iron uptake and . PA also translocated the tfR1 by stimulating calcium influx, but the calcium chelator, BAPTA-AM, dramatically reduced iron overload by inhibiting tfR1 translocation and ultimately increased sensitivity. Iron overload may play a critical role in PA-induced . Blocking iron overload may thus be a useful strategy for preventing and diabetes.-Cui, R., Choi, S.-E., Kim, T. H., Lee, H. J., Lee, S. J., Kang, Y., Jeon, J. Y., Kim, H. J., Lee, K.-W. Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced in human skeletal muscle cells.

Keyword: insulin resistance

Interesterified Palm Olein (IEPalm) and Interesterified Stearic -Rich Fat Blend (IEStear) Have No Adverse Effects on : A Randomized Control Trial.

Chemically-interesterified (CIE) fats are -fat free and are increasingly being used as an alternative to hydrogenated oils for food manufacturing industries to optimize their products\' characteristics and nutrient compositions. The metabolic effects of CIE fats on activity, lipids, and adiposity in humans are not well established. We investigated the effects of CIE fats rich in (C16:0, IEPalm) and stearic (C18:0, IEStear) acids on , serum lipids, apolipoprotein concentrations, and adiposity, using C16:0-rich natural palm olein (NatPO) as the control. We designed a parallel, double-blind clinical trial. Three test fats were used to prepare daily snacks for consumption with a standard background diet over a period of 8 weeks by three groups of a total of 85 healthy, overweight adult volunteers. We measured the outcome variables at weeks 0, 6, and at the endpoint of 8. After 8 weeks, there was no significant difference in surrogate biomarkers of in any of the IE fat diets (IEPalm and IEStear) compared to the NatPO diet. The change in serum triacylglycerol concentrations was significantly lower with the IEStear diet, and the changes in serum leptin and body fat percentages were significantly lower in the NatPO-diet compared to the IEPalm diet. We conclude that diets containing C16:0 and C18:0-rich CIE fats do not affect markers of compared to a natural C16:0-rich fat (NatPO) diet. Higher amounts of saturated fatty acids (SFAs) and longer chain SFAs situated at the -1,3 position of the triacylglycerol (TAG) backbones resulted in less weight gain and lower changes in body fat percentage and leptin concentration to those observed in NatPO and IEStear.

Keyword: insulin resistance

The fatty -rich fraction of Eruca sativa (rocket salad) leaf extract exerts antidiabetic effects in cultured skeletal muscle, adipocytes and liver cells.

Eruca sativa Mill. (Brassicaceae), commonly known as rocket salad, is a popular leafy-green vegetable with many health benefits.To evaluate the antidiabetic activities of this plant in major -responsive tissues.Five E. sativa leaf extracts of varying polarity were prepared (aqueous extract, 70% and 95% ethanol extracts, the n-hexane-soluble fraction of the 95% ethanol extract (ES3) and the defatted 95% ethanol extract). Eruca sativa extracts were investigated through a variety of cell-based in vitro bioassays for antidiabetic activities in C2C12 skeletal muscle cells, H4IIE hepatocytes and 3T3-L1 adipocytes. Guided by the results of these bioassays, ES3 was fractionated into the saponifiable (SM) and the unspaonifiable (USM) fractions. Glucose uptake was measured using [H]-deoxy-glucose, while the effects on hepatic glucose-6-phosphatase (G6Pase) and adipogenesis were assessed using Wako AutoKit Glucose and AdipoRed assays, respectively.ES3 and its SM fraction significantly stimulated glucose uptake with EC values of 8.0 and 5.8\u2009μg/mL, respectively. Both extracts significantly inhibited G6Pase activity (IC values of 4.8 and 9.3\u2009μg/mL, respectively). Moreover, ES3 and SM showed significant adipogenic activities with EC of 4.3 and 6.1\u2009μg/mL, respectively. Fatty content of SM was identified by GC-MS. trans-Vaccenic and palmitoleic acids were the major unsaturated fatty acids, while and azelaic acids were the main saturated fatty acids.These findings indicate that ES3 and its fatty -rich fraction exhibit antidiabetic activities in -responsive cell lines and may hence prove useful for the treatment of type 2 diabetes.

Keyword: insulin resistance

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of obesity, , type-2 diabetes and metabolic syndrome. Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal metabolic disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA metabolism on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: insulin resistance

The effect of FADS2 gene rs174583 polymorphism on desaturase activities, fatty profile, , biochemical indices, and incidence of type 2 diabetes.

In this study, we investigated the associations of erythrocytes fatty composition, activities of delta-5 desaturase (D5D) and delta-6 desaturase (D6D), and other metabolic risk factors, with type 2 diabetes (T2D) risk to determine if rs174583 polymorphism of FADS2 gene had any effect on these associations.Fatty profile of erythrocytes was determined using gas chromatography-mass spectrometry in 95 T2D patients and 95 apparently healthy participants. The genotypes of single-nucleotide polymorphism (SNP) of FADS2 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism technique. Other biochemical parameters were measured in the serum using standard analytical procedures.D6D activity was increased ( < 0.001) and D5D activity was decreased in T2D patients ( < 0.001) compared to controls. Homeostatic model assessment (HOMA-IR) index was positively correlated with D6D ( = 0.34, < 0.001) and negatively correlated with D5D ( = -0.19, = 0.02). ( < 0.001) and dihomo-gamma-linolenic ( = 0.03) were higher and linoleic ( < 0.001) and arachidonic (AA) ( < 0.001) were lower in T2D patients. The distribution of rs174583 genotypes which includes C/T, C/C, and T/T was not different in the two groups ( = 0.63).In the population studied, there was a strong association in the erythrocytes fatty composition, D5D and D6D activities and other metabolic risk factors between non-T2D and T2D patients. In addition, there was a strong association in erythrocytes DGLA and AA contents and D5D activities between rs174583 genotypes in all participants. However, the distribution of rs174583 genotypes did not differ significantly between T2D patient and controls, and it did not appear to be an association between rs174583 SNP and incident of T2D.

Keyword: insulin resistance

Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High-Fat-Diet-Induced Cardiac Dysfunction and by Elevating Tafazzin in Mouse Hearts.

High fat (HF)-diet-induced is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low fat (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the mitochondria. Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to via mediating Drp-1 translocation to the mitochondria, and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates .© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: insulin resistance

Preventive effect of oleate on palmitate-induced in skeletal muscle and its mechanism of action.

in skeletal muscle is a feature associated with exposure to an excess of saturated fatty acids such as palmitate. Oleic has been shown to blunt palmitate-induced in muscle cells. However, there is no literature available regarding the effect of oleic on palmitate-induced in intact muscle. Therefore, this study investigated the effect of oleic on palmitate-induced in rat soleus muscle and its underlying mechanisms. For these purposes, oleic (1\xa0mM) was administered for 12\xa0h in the absence or presence of palmitate (2\xa0mM). At the end of the experiment, plasmalemmal GLUT4, the phosphorylation of AS160 and Akt-2, and the total expression of these signaling proteins were examined. We found that treatment with palmitate for 12\xa0h reduced -stimulated GLUT4 translocation and the phosphorylation of AS160 and Akt-2. However, the administration of oleic fully restored -stimulated GLUT4 translocation (P\xa0<\xa00.05), as well as AS160 and Akt-2 phosphorylation (P\xa0<\xa00.05) despite the continuous presence of palmitate. Wortmannin, an inhibitor of PI3-K, only slightly prevented the oleic -induced improvements in -stimulated GLUT4 translocation, and AS160 phosphorylation. However, this treatment completely inhibited the oleic -induced improvement in -stimulated Akt-2 phosphorylation. In contrast, the oleic -induced improvement in signaling was not affected by compound C, an AMPK specific inhibitor. In conclusion, the results clearly indicate that oleic administration alleviates palmitate-induced by promoting GLUT4 translocation in muscle, at least in part, by activating the PI3K pathway.

Keyword: insulin resistance

Oxidative stress induced by modulates K2.3 channels in vascular endothelium.

Elevated plasma free fatty acids level has been implicated in the development of , inflammation, and endothelial dysfunction in diabetic and nondiabetic individuals. However, the underlying mechanisms still remain to be defined. Herein, we investigated the effect of (PA), the most abundant saturated fatty in the human body, on small-conductance Ca-activated potassium channels (K2.3)-mediated relaxation in rodent arteries and the underlying molecular mechanism. The effect of PA on K2.3 in endothelium was evaluated using real-time PCR, Western blotting, whole-cell patch voltage-clamp, wire and pressure myograph system, and reactive oxygen species (ROS) were measured by using dihydroethidium and 2\', 7\'-dichlorofluorescein diacetate. K2.3-mediated vasodilatation responses to acetylcholine and NS309 (agonist of K2.3 and K3.1) were impaired by incubation of normal mesenteric arteries with 100\u202fμM\u202fPA for 24\u202fh. In cultured human umbilical vein endothelial cells (HUVECs), PA decreased K2.3 current and expression at mRNA and protein levels. Incubation with the NADPH oxidase (Nox) inhibitor dibenziodolium (DPI) partly inhibited the PA-induced ROS production and restored K2.3 expression. Inhibition of either p38-MAPK or NF-κB using specific inhibitors (SB203580, SB202190 or Bay11-7082, pyrrolidinedithiocarbamate) attenuated PA-induced downregulation of K2.3 and inhibition of p38-MAPK also attenuated PA-induced phosphorylation of NF-κB p65. Furthermore, DPI reversed the increment of phospho-p38-MAPK by PA. These results demonstrated that PA downregulated K2.3 expressions via Nox/ROS/p38-MAPK/NF-κB signaling leading to endothelial vasodilatory dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and .We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, , leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, , leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: insulin resistance

Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated and hepatic glucogenesis, regulated serum lipid parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

PPARγ alleviated hepatocyte steatosis through reducing SOCS3 by inhibiting JAK2/STAT3 pathway.

Peroxisome proliferator-activated receptor gamma (PPARγ) participates in the process of (IR), a crucial pathophysiology in non-alcoholic fatty liver disease (NAFLD). Meanwhile, suppressor of cytokine signaling3 (SOCS3) also regulates IR in NAFLD. Both PPARγ and SOCS3 play a role in NAFLD through regulating IR, while it is unclear whether these two proteins interact to regulate hepatic steatosis. PPARγ, SOCS3 and its associated JAK2/STAT3 pathway were analyzed using Kuppfer cells (KCs) treatment with LPS and BRL-3A cells treatment with , KC-conditioned medium (KCCM), PPARγ agonist rosiglitazone (ROZ) or JAK2 inhibitor AG490 to demonstrate the role of PPARγ and SOCS3 in hepatocytes steatosis. As LPS concentration increasing, phagocytosis activity of KCs decreased; but releasing of TNF-α and IL-6 increased. After treatment with KCCM, mRNA level of SOCS3, JAK2 and STAT3 as well as protein expression of SOCS3, p-JAK2 and p-STAT3 in steatosis BRL-3A cells increased significantly, which were inhibited by AG490 or ROZ treatment. Taken together, these results indicated that KCCM attributed to KCs dysfunction facilitated hepatocyte steatosis through promoting expressing SOCS3; but PPARγ agonist ROZ alleviated steatosis through reducing SOCS3 expression by inhibiting JAK2/STAT3 in hepatocytes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: insulin resistance

Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4-MD2 complex.

Reactive oxygen species (ROS) contribute to the development of non-alcoholic fatty liver disease. ROS generation by infiltrating macrophages involves multiple mechanisms, including Toll-like receptor 4 (TLR4)-mediated NADPH oxidase (NOX) activation. Here, we show that palmitate-stimulated CD11bF4/80 hepatic infiltrating macrophages, but not CD11bF4/80 Kupffer cells, generate ROS via dynamin-mediated endocytosis of TLR4 and NOX2, independently from MyD88 and TRIF. We demonstrate that differently from LPS-mediated dimerization of the TLR4-MD2 complex, palmitate binds a monomeric TLR4-MD2 complex that triggers endocytosis, ROS generation and increases pro-interleukin-1β expression in macrophages. Palmitate-induced ROS generation in human CD68CD14 macrophages is strongly suppressed by inhibition of dynamin. Furthermore, Nox2-deficient mice are protected against high-fat diet-induced hepatic steatosis and . Therefore, endocytosis of TLR4 and NOX2 into macrophages might be a novel therapeutic target for non-alcoholic fatty liver disease.

Keyword: insulin resistance

Epigallocatechin gallate improves in HepG2 cells through alleviating inflammation and lipotoxicity.

High levels of circulating free fatty acids (FFAs), inflammation and oxidative stress are important causes for (IR) and type 2 diabetes mellitus. The aim of this study was to investigate the mechanisms of EGCG in alleviating IR in HepG2 cells.HepG2 cells were treated with 25\u202fmM glucose, 0.25\u202fmM (PA), or 50\u202fμM EGCG for 24\u202fh.EGCG increased glucose uptake and decreased glucose content. EGCG markedly decreased the levels of inflammatory and oxidative stress factors including nuclear factor κB (NF-κB), tumor necrosis factor-α, interleukin-6, reactive oxygen species, malondialdehyde and p53 protein, and markedly increased superoxide dismutases (SOD), glutathione peroxidase and SOD2 protein. EGCG significantly downregulated the levels of FFAs, triacylglycerol and cholesterol in HepG2 cells. The glucose transporter 2 (GLUT2) protein and its downstream proteins peroxisome proliferator-activated receptor γ coactivator (PGC)-1β were significantly increased, and sterol regulatory element-binding-1c (SREBP-1c) protein, and fatty synthase (FAS) were significantly decreased by EGCG in HepG2. Moreover, the foregoing effects were reversed by siRNA-mediated knockdown of GLUT2.Our data demonstrated that EGCG improved IR, possibly through ameliorating glucose (25\u202fmM) and PA (0.25\u202fmM)-induced inflammation, oxidative stress, and FFAs via the GLUT2/PGC-1β/SREBP-1c/FAS pathway in HepG2 cells.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

AMP-activated protein kinase-mediated expression of heat shock protein beta 1 enhanced sensitivity in the skeletal muscle.

Activation of AMP-activated protein kinase (AMPK) has been viewed as an important target for the treatment of . Here, by proteomic analysis, we found that expression of heat shock protein beta-1 (HSPB1) was induced by the AMP analog 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside in palmitate-induced -resistant cells. Overexpression of AMPKα2, or activation of AMPKα via acute/chronic exercise training, increased HSPB1 expression in the skeletal muscle. In AMPKα2 mice, HSPB1 expression was downregulated in the quadriceps muscles. Exercise did not increase HSPB1 expression in AMPKα2 mice. Moreover, overexpression of HSPB1 enhanced sensitivity in palmitate-induced -resistant cells and restored metabolic phenotypes associated with defective AMPK. Finally, HSPB1 was required for AMPK-mediated activation of the class IIa histone deacetylases and glucose uptake in the skeletal muscle. Our results demonstrate that AMPK-mediated HSPB1 expression enhanced sensitivity in the skeletal muscle.© 2016 Federation of European Biochemical Societies.

Keyword: insulin resistance

A novel PPARα/γ agonist, propane-2-sulfonic octadec-9-enyl-amide, ameliorates and gluconeogenesis in vivo and vitro.

Peroxisome proliferator-activated receptor alpha/gamma (PPARα/γ) agonists have emerged as important pharmacological agents for improving action. Propane-2-sulfonic octadec-9-enyl-amide (N15) is a novel PPARα/γ dual agonist synthesized in our laboratory. The present study investigates the efficacy and safety of N15 on regulation in high fat diet (HFD)-and streptozotocin (STZ)-induced diabetic mice and in (PA)-induced HepG2 cells. Our results showed that N15 remarkably ameliorated and dyslipidemia in vivo, as well as rectified the glucose consumption and gluconeogenesis in vitro. Moreover, the glucose-lowering effect of N15 was associated with PPARγ mediated up-regulation of hepatic glucose consumption and down-regulation of gluconeogenesis. Meanwhile, N15 exerted advantageous effects on glucose and lipid metabolism without triggering weight gain and hepatotoxicity in mice. In conclusion, our data demonstrated that by alleviating glucose and lipid abnormalities, N15 could be used as a potential prophylactic and therapeutic agent against type 2 diabetes and related metabolic disorders.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: insulin resistance

Myricanol modulates skeletal muscle-adipose tissue crosstalk to alleviate high-fat diet-induced obesity and .

Skeletal muscle is the predominant site for glucose disposal and fatty consumption. Emerging evidence indicates that the crosstalk between adipose tissue and skeletal muscle is critical in maintaining sensitivity and lipid homeostasis. The current study was designed to investigate whether myricanol improves sensitivity and alleviates adiposity through modulating skeletal muscle-adipose tissue crosstalk.The therapeutic effect of myricanol was evaluated on (PA)-treated C2C12 myotubes and high-fat diet (HFD)-fed mice. The crosstalk between myotubes and adipocytes was evaluated using Transwell assay. The cellular lipid content was examined by Nile red staining. The mitochondrial content was assessed by MitoTracker Green staining and citrate synthase activity, and the mitochondrial function was examined by Seahorse assay. Expression of mitochondria-related and signalling pathway proteins was analysed by Western blot, and the irisin level was determined by elisa kit.Myricanol increased mitochondrial quantity and function through activating AMP-activated protein kinase, resulting in reduced lipid accumulation and enhanced -stimulated glucose uptake, in PA-treated C2C12 myotubes. Furthermore, myricanol stimulated irisin production and secretion from myotubes to reduce lipid content in 3T3-L1 adipocytes. In HFD-fed mice, myricanol treatment alleviated adiposity and through enhancing lipid utilization and irisin production in skeletal muscle and inducing browning of inguinal fat.Myricanol modulates skeletal muscle-adipose tissue crosstalk, to stimulate browning of adipose tissue and improve sensitivity in skeletal muscle. Myricanol might be a potential candidate for treating and obesity.© 2019 The British Pharmacological Society.

Keyword: insulin resistance

Disruption of Lipid Uptake in Astroglia Exacerbates Diet-Induced Obesity.

Neuronal circuits in the brain help to control feeding behavior and systemic metabolism in response to afferent nutrient and hormonal signals. Although astrocytes have historically been assumed to have little relevance for such neuroendocrine control, we investigated whether lipid uptake via lipoprotein lipase (LPL) in astrocytes is required to centrally regulate energy homeostasis. Ex vivo studies with hypothalamus-derived astrocytes showed that LPL expression is upregulated by oleic , whereas it is decreased in response to or triglycerides. Likewise, astrocytic LPL deletion reduced the accumulation of lipid droplets in those glial cells. Consecutive in vivo studies showed that postnatal ablation of LPL in glial fibrillary acidic protein-expressing astrocytes induced exaggerated body weight gain and glucose intolerance in mice exposed to a high-fat diet. Intriguingly, astrocytic LPL deficiency also triggered increased ceramide content in the hypothalamus, which may contribute to hypothalamic . We conclude that hypothalamic LPL functions in astrocytes to ensure appropriately balanced nutrient sensing, ceramide distribution, body weight regulation, and glucose metabolism.© 2017 by the American Diabetes Association.

Keyword: insulin resistance

Polymorphism in the Gene Is Associated to Changes in Fatty Metabolism and Circulating Endocannabinoid Levels Distinctively in Normal Weight and Obese Subjects.

The multifunctional CD36 scavenger receptor facilitates fatty (FA) uptake and oxidation and it has been involved in the pathophysiology related to dysfunctional FA metabolism. The common variant in the gene, (A/G), whose allele A is characterized by a reduced protein expression, has been associated with taste sensitivity to and preference for fat. We therefore aimed at evaluating whether the polymorphism may influence fatty metabolism and endocannabinoid biosynthesis in normal weight (NW) and obese (OB) subjects. Red blood cell (RBC) fatty composition, and plasma endocannabinoid levels were determined. In NW subjects with AA genotype was found a marked reduction of RBC saturated fatty acids and /linoleic ratio (PA/LA), considered as (DNL) biomarkers. Remarkably, to the reduction of DNL biomarkers corresponded an increase of omega-6 index, an indirect marker of the impact on fatty metabolism of dietary omega-6 fatty acids, endocannabinoid levels and a higher waist/hip ratio. The presence of the G allele was instead associated with increased endocannabinoid plasma levels and a trend for increased waist/hip ratio in obese subjects, even though exhibited decreased BMI with respect to those with AA genotype. These data indicate that the polymorphism, , leads to a distinct metabolic pattern in NW and in OB subjects. Therefore, their determination may be crucial in developing personalized therapeutic strategies for ameliorating dyslipidemia and other metabolic disorders.

Keyword: lipogenesis

Trigonelline prevents high cholesterol and high fat diet induced hepatic lipid accumulation and lipo-toxicity in C57BL/6J mice, via restoration of hepatic autophagy.

Non-alcoholic fatty liver disease (NAFLD) is often linked with impaired hepatic autophagy. Here, we studied the alterations in hepatocellular autophagy by high cholesterol and high-fat diet (HC-HF) diet in C57BL/6J mice, and by (PA), in AML-12 and HepG2 cells. Further, we analysed role of Trigonelline (TG), a plant alkaloid, in preventing NAFLD, by modulating autophagy. For this, C57BL/6J mice were fed with Standard Chow (SC) or HC-HF diet, with and without TG for 16 weeks. In-vitro; AML-12\u202fcells and HepG2 cells, were exposed to PA with and without TG, for 24\u202fh. Cellular events related to autophagy, , and lipo-toxicity were studied. The HC-HF diet fed mice showed hepatic autophagy blockade, increased triglycerides and steatosis. PA exposure to AML-12\u202fcells and HepG2 cells induced impaired autophagy, ER stress, resulting in lipotoxicity. TG treatment in HC-HF fed mice, restored hepatic autophagy, and prevented steatosis. TG treated AML-12, and HepG2 cells exposed to PA showed autophagy restoration, and reduced lipotoxicity, however, these effects were diminished in Atg7-/- HepG2 cells, and in the presence of chloroquine. This study shows that HC-HF diet-induced impaired autophagy, and steatosis is prevented by TG, which attributes to its novel mechanism in treating NAFLD.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: lipogenesis

Development of an in vitro model to study hepatitis C virus effects on hepatocellular lipotoxicity and lipid metabolism.

Hepatic steatosis is common in patients infected with hepatitis C virus (HCV). Particularly in patients infected with non-genotype 3 HCV, hepatic steatosis is closely related to factors of the metabolic syndrome such as hyperlipidemia. However, the molecular mechanisms involved in this "metabolic" steatosis in non-3 genotype HCV infections are not well understood. Here, we aimed to develop an in vitro model to study the effect of genotype 1 HCV infection on hepatic lipotoxicity and lipid metabolism. Cellular lipid accumulation was induced in Huh-7 hepatoma cells transfected with HCV genotype 1b replicon (HCV) by incubation with increasing doses of (C16:0) or oleic (C18:1 n-9) complexed to albumin mimicking hyperlipidemic conditions. Mock transfected hepatoma cells (HCV) were used as controls. Incubation with oleic concentrations as high as 0.5\u202fmM did not induce toxic effects in HCV or HCV cells. In contrast, incubation with caused dose-dependently cytotoxic effects which were more pronounced in HCV compared to HCV cells. Further analysis with subtoxic and oleic concentrations revealed a higher uptake of fatty acids and intracellular triglyceride accumulation in HCV compared to HCV cells. Carnitine palmitoyltransferase I (CPT1) expression, indicative of mitochondrial beta-oxidation, was markedly stimulated by lipid exposure in HCV but not in HCV cells. Furthermore, heme oxygenase 1 (HMOX1) expression levels increased in FA stimulated cells, and this increase was significantly higher in HCV compared to HCV cells. In contrast, expression of the key enzymes of hepatic de novo fatty synthase (FASN) and stearoyl-CoA desaturase (SCD-1) was significantly reduced upon oleate exposure in HCV but not in HCV cells. In summary, our newly developed cell culture model revealed effects of HCV genotype 1b infection on metabolic susceptibility to lipid accumulation and toxicity particularly to saturated lipids. These results may indicate that HCV (genotype 1b) infected individuals with hyperlipidemia may benefit from dietary or pharmacological intervention.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: lipogenesis

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on lipid metabolism and inflammation in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular lipid contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet lipid metabolic disorder and islet inflammation in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: lipogenesis

Differential capability of metabolic substrates to promote hepatocellular lipid accumulation.

Excessive storage of triacylglycerides (TAGs) in lipid droplets within hepatocytes is a hallmark of non-alcoholic fatty liver disease (NAFLD), one of the most widespread metabolic disorders in Western societies. For the purpose of exploring molecular pathways in NAFLD development and testing potential drug candidates, well-characterised experimental models of ectopic TAG storage in hepatocytes are needed.Using an optimised Oil Red O assay, immunoblotting and real-time qRT-PCR, we compared the capability of dietary monosaccharides and fatty acids to promote lipid accumulation in HepG2 human hepatoma cells.Both high glucose and high fructose resulted in intracellular lipid accumulation after 48\xa0h, and this was further augmented (up to twofold, as compared to basal levels) by co-treatment with the -stimulating hormone insulin and the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-α), respectively. The fatty acids and oleic were even more effective than these carbohydrates, inducing significantly elevated TAG storage already after 24\xa0h of treatment. Highest (about threefold) increases in lipid accumulation were observed upon treatment with oleic , alone as well as in combinations with or with high glucose and insulin. Increases in protein levels of a major lipid droplet coat protein, perilipin-2 (PLIN2), mirrored intracellular lipid accumulation following different treatment regimens.Several treatment regimens of excessive fat and sugar supply promoted lipid accumulation in HepG2 cells, albeit with differences in the extent and rapidity of steatogenesis. PLIN2 is a candidate molecular marker of sustained lipid accumulation in HepG2 cells.

Keyword: lipogenesis

Ncb5or deficiency increases fatty catabolism and oxidative stress.

The endoplasmic reticulum-associated NADH cytochrome b(5) oxidoreductase (Ncb5or) is widely distributed in animal tissues. Ncb5or(-/-) mice develop diabetes at age 7 weeks and have increased susceptibility to the diabetogenic oxidant streptozotocin. Ncb5or deficiency also results in lipoatrophy and increased hepatocyte sensitivity to cytotoxic effects of saturated fatty acids. Here we investigate the mechanisms of these phenomena in prediabetic Ncb5or(-/-) mice and find that, despite increased rates of fatty uptake and synthesis and higher stearoyl-CoA desaturase (SCD) expression, Ncb5or(-/-) liver accumulates less triacylglycerol (TAG) than wild type (WT). Increased fatty catabolism and oxidative stress are evident in Ncb5or(-/-) hepatocytes and reflect increased mitochondrial content, peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) expression, fatty oxidation rates, oxidative stress response gene expression, and oxidized glutathione content. Ncb5or(-/-) hepatocytes readily incorporate exogenous fatty acids into TAG but accumulate more free fatty acids (FFA) and have greater palmitate-induced oxidative stress responses and cell death than WT, all of which are alleviated by co-incubation with oleate via TAG channeling. A high fat diet rich in palmitate and oleate stimulates both and fatty catabolism in Ncb5or(-/-) liver, resulting in TAG levels similar to WT but increased intracellular FFA accumulation. Hepatic SCD-specific activity is lower in Ncb5or(-/-) than in WT mice, although Ncb5or(-/-) liver has a greater increase in Scd1 mRNA and protein levels. Together, these findings suggest that increased FFA accumulation and catabolism and oxidative stress are major consequences of Ncb5or deficiency in liver.

Keyword: lipogenesis

Subcutaneous adipose tissue fatty desaturation in adults with and without rare adipose disorders.

Elevated stearoyl-CoA desaturase activity has been described in obese states, with an increased desaturation index (DI) suggesting enhanced . Differences in the DI among various phenotypes of abnormal adiposity have not been studied. Abnormal accumulation of subcutaneous adipose tissue occurs in rare adipose disorders (RADs) including Dercum\'s disease (DD), multiple symmetric lipomatosis (MSL), and familial multiple lipomatosis (FML). Examining the DI in subcutaneous fat of people with DD, MSL and FML may provide information on adipose tissue fatty metabolism in these disorders. The aims of this pilot study were: 1) to determine if differences in adipose tissue DIs are present among RADs, and 2) to determine if the DIs correlate to clinical or biochemical parameters.Subcutaneous adipose tissue was obtained from human participants with DD (n = 6), MSL (n = 5), FML (n = 8) and obese Controls (n = 6). Fatty composition was determined by gas chromatography/mass spectrometry. The DIs (palmitoleic/, oleic/stearic, vaccenic/stearic ratios) were calculated from the gas chromatogram peak intensities. SCD1 gene expression was determined. Spearman\'s correlations between the DIs and available clinical or biochemical data were performed.In DD subjects, the vaccenic/stearic index was lower (p < 0.05) in comparison to Controls. Percent of total of the saturated fatty myristic was higher in DD compared with Controls and FML. Percent of monounsaturated vaccenic in DD trended lower when compared with Controls, and was decreased in comparison to FML. In MSL, total percent of the polyunsaturated fatty acids was significantly lower than in the Control group (p < 0.05). In the total cohort of subjects, the palmitoleic/ and oleic/stearic DIs positively correlated with age, BMI, and percent body fat.The positive associations between the DIs and measures of adiposity (BMI and percent body fat) support increased desaturase activity in obesity. The lower vaccenic/stearic DI in DD SAT compared with Controls suggests presence of other factors involved in fat accumulation in addition to lifestyle. Other mechanisms driving fat accumulation in DD such as inflammation or lymphatic dysfunction should be investigated.© 2012 Yee et al; licensee BioMed Central Ltd.

Keyword: lipogenesis

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model.

Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free fatty -induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free fatty acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.

Keyword: lipogenesis

Does the enterolactone (ENL) affect fatty transporters and lipid metabolism in liver?

NAFLD as a result of inappropriate diet and obesity, may progress to sever conditions such as: type 2 diabetes mellitus or steatohepatitis, and has recently become a prevalent topic of numerous investigations. Due to its dangerous aftermaths, finding new substances, such as polyphenols and their derivatives, which might reduce liver steatosis is the main target of research into NAFLD treatment. Hence, the aim of the present study was to evaluate the effect(s) of enterolactone (ENL), a metabolite of secoisolariciresinol (SECO), on lipid metabolism together with changes in the expression of fatty transporters in fatty liver.The experiments were conducted on HepG2 cells incubated with either ENL and/or during 16\xa0h exposure. The expression of selected fatty transport proteins: FATP2, FATP5, CD36, FABPpm, ABCA1, MTP, ACBP and L-FABP, as well as the proteins directly involved in lipogenesis (FAS), oxidation pathway (CPT 1), and lipid metabolism (PPARα, LXR, SREBP1c, pAMPK) was estimated by Western Blot. Intra and extracellular lipid contents were assessed by Gas-Liquid Chromatography. The data was analyzed with two-way analysis of variance (ANOVA), and results were considered to be statistically significant at \xa0≤\xa00.05.ENL stimulated extracellular efflux of free fatty acids (FFA) and triacylglicerols (TAG) to the medium, while, it had no influence on FATP-family mediated intracellular fatty uptake. Moreover, ENL decreased the expression of CPT 1, pAMPK, PPARα, increased SREBP1c and had no effect on LXR, and FAS content.The findings of our study demonstrate that ENL had opposite effect on liver steatosis in comparison with other polyphenols what suggests that it may be an inactive metabolite. ENL did not affect significantly the intracellular accumulation of FFA, DAG and TAG, yet it promoted their extracellular efflux. Furthermore, it inhibited ß-oxydation and intracellular lipid metabolism what may contribute to the progression of NAFLD.

Keyword: lipogenesis

Absence of Carbohydrate Response Element Binding Protein in Adipocytes Causes Systemic Insulin Resistance and Impairs Glucose Transport.

Lower adipose-ChREBP and de novo (DNL) are associated with insulin resistance in humans. Here, we generated adipose-specific ChREBP knockout (AdChREBP KO) mice with negligible sucrose-induced DNL in adipose tissue (AT). Chow-fed AdChREBP KO mice are insulin resistant with impaired insulin action in the liver, muscle, and\xa0AT and increased AT inflammation. HFD-fed AdChREBP KO mice are also more insulin resistant\xa0than controls. Surprisingly, adipocytes lacking ChREBP display a cell-autonomous reduction in insulin-stimulated glucose transport that is mediated by impaired Glut4 translocation and exocytosis, not lower Glut4 levels. AdChREBP KO mice have lower levels of esters of hydroxy stearic acids (PAHSAs) in serum, and AT. 9-PAHSA supplementation completely rescues their insulin resistance and AT inflammation. 9-PAHSA also normalizes impaired glucose transport and Glut4 exocytosis in ChREBP KO adipocytes. Thus, loss\xa0of adipose-ChREBP is sufficient to cause insulin resistance, potentially by regulating AT glucose transport and flux through specific lipogenic pathways.Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Genetic variation in genes of the fatty synthesis pathway and breast cancer risk.

Fatty synthase (FAS) is the major enzyme of . It catalyzes the NADPH-dependent condensation of acetyl-CoA and malonyl-CoA to produce . Transcription of the FAS gene is controlled synergistically by the transcription factors ChREBP (carbohydrate response element-binding protein), which is induced by glucose, and SREBP-1 (sterol response element-binding protein-1), which is stimulated by insulin through the PI3K/Akt signal transduction pathway. We investigated whether the genetic variability of the genes encoding for ChREBP, SREBP and FAS (respectively, MLXIPL, SREBF1 and FASN) is related to breast cancer risk and body-mass index (BMI) by studying 1,294 breast cancer cases and 2,452 controls from the European Prospective Investigation on Cancer (EPIC). We resequenced the FAS gene and combined information of SNPs found by resequencing and SNPs from public databases. Using a tagging approach and selecting 20 SNPs, we covered all the common genetic variation of these genes. In this study we were not able to find any statistically significant association between the SNPs in the FAS, ChREBP and SREPB-1 genes and an increased risk of breast cancer overall and by subgroups of age, menopausal status, hormone replacement therapy (HRT) use or BMI. On the other hand, we found that two SNPs in FASN were associated with BMI.

Keyword: lipogenesis

Beyond the Scavenging of Reactive Oxygen Species (ROS): Direct Effect of Cerium Oxide Nanoparticles in Reducing Fatty Acids Content in an In Vitro Model of Hepatocellular Steatosis.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic accumulation of lipids. Antisteatotic effects of cerium oxide nanoparticles (CeONPs) have recently been shown in animal models of liver disease. However, it is unclear whether the activity of CeONPs is related solely to the decrease in oxidative stress or, in addition, they directly decrease liver fatty accumulation. To address this question, in this work, we used an in vitro model of hepatocellular steatosis, exposing HepG2 cells to oleic and . Cell uptake of CeONPs and their effect on oxidative stress and viability of hepatic cells cultured with HO were also evaluated. Results show that CeONPs were uptaken by HepG2 cells and reduced oxidative stress and improved cell viability. Treatment with oleic and increased and the content of different fatty acids. CeONPs reduced and stearic and most fatty acids consisting of more than 18 carbon atoms. These effects were associated with significant changes in elongase and desaturase activity. In conclusion, CeONPs directly protected HepG2 cells from cell injury in oxidative stress conditions and reduced fatty content in steatotic conditions by inducing specific changes in fatty metabolism, thus showing potential in the treatment of NAFLD.

Keyword: lipogenesis

Dietary Fatty Acids Differentially Regulate Secretion of Adiponectin and Interleukin-6 in Primary Canine Adipose Tissue Culture.

The aim of this study was to determine the effect of n3 polyunsaturated fatty acids (PUFA) on canine adipose tissue secretion of adiponectin, interleukin-6 (IL6), and tumor necrosis factor-α (TNFα). Subcutaneous and omental visceral adipose tissue samples were collected from 16 healthy intact female dogs. Concentrations of adiponectin were measured in mature adipocyte cultures, and concentrations of IL6 and TNFα were measured in undifferentiated stromovascular cell (SVC) cultures following treatment with eicosapentaenic (EPA, 20:5n-3), arachidonic (ARA, 20:4n-6), or (PAM, 16:0) at 25, 50, or 100\u2009μM. Secretion of adiponectin from mature adipocytes was higher (p < 0.001) following EPA treatment at 50\u2009μM compared to control in subcutaneous tissue, and higher following EPA treatment compared to PAM treatment at 25\u2009μM in both subcutaneous (p < 0.001) and visceral tissues (p = 0.010). Secretion of IL6 from SVC derived from subcutaneous tissue was lower following EPA treatment and higher following PAM treatment compared to control both at 50\u2009μM (p = 0.001 and p = 0.041, respectively) and 100\u2009μM (p = 0.013 and p < 0.001, respectively). These findings of stimulation of adiponectin secretion and inhibition of IL6 secretion by EPA, and stimulation of IL6 secretion by PAM, are consistent with findings of increased circulating concentrations of adiponectin and decreased circulating concentration of IL6 in dogs supplemented with dietary fish oil, and show that the effect of fish oil on circulating concentrations of adiponectin and IL6 is, at least partially, the result of local effects of EPA and PAM on adipose tissue.© 2018 AOCS.

Keyword: lipogenesis

Picroside II attenuates fatty accumulation in HepG2 cells via modulation of fatty uptake and synthesis.

Hepatic steatosis is caused by an imbalance between free fatty acids (FFAs) uptake, utilization, storage, and disposal. Understanding the molecular mechanisms involved in FFAs accumulation and its modulation could drive the development of potential therapies for Nonalcoholic fatty liver disease. The aim of the current study was to explore the effects of picroside II, a phytoactive found in , on fatty accumulation vis-à-vis silibinin, a known hepatoprotective phytoactive from .HepG2 cells were loaded with FFAs (oleic :/2:1) for 20 hours to mimic hepatic steatosis. The FFAs concentration achieving maximum fat accumulation and minimal cytotoxicity (500 μM) was standardized. HepG2 cells were exposed to the standardized FFAs concentration with and without picroside II pretreatment.Picroside II pretreatment inhibited FFAs-induced lipid accumulation by attenuating the expression of fatty transport protein 5, sterol regulatory element binding protein 1 and stearoyl CoA desaturase. Preatreatment with picroside II was also found to decrease the expression of forkhead box protein O1 and phosphoenolpyruvate carboxykinase.These findings suggest that picroside II effectively attenuated fatty accumulation by decreasing FFAs uptake and lipogenesis. Picroside II also decreased the expression of gluconeogenic genes.

Keyword: lipogenesis

An Essential Role for the Tumor-Suppressor Merlin in Regulating Fatty Synthesis.

Neurofibromatosis type 2 (NF2) is an autosomal dominant disorder characterized by the development of multiple tumors in the central nervous system, most notably schwannomas, and meningiomas. Mutational inactivation of the gene encoding the protein Merlin is found in most sporadic and inherited schwannomas, but the molecular mechanisms underlying neoplastic changes in schwannoma cells remain unclear. We report here that Nf2-deficient cells display elevated expression levels of key enzymes involved in and that this upregulation is caused by increased activity of Torc1. Inhibition or knockdown of fatty synthase (FASN), the enzyme that catalyzes the formation of from malonyl-CoA, drove -deficient cells into apoptosis. Treatment of -mutant cells with agents that inhibit the production of malonyl-CoA reduced their sensitivity to FASN inhibitors. Collectively, these results suggest that the altered lipid metabolism found in -mutant cells renders them sensitive to elevated levels of malonyl-CoA, as occurs following blockade of FASN, suggesting new targeted strategies in the treatment of -deficient tumors. .©2017 American Association for Cancer Research.

Keyword: lipogenesis

GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic fatty homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced liver dysfunction, hepatic steatosis and insulin resistance. In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and , and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), fatty synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the liver may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Overexpression of juxtaposed with another zinc finger gene 1 reduces proinflammatory cytokine release via inhibition of stress-activated protein kinases and nuclear factor-κB.

As an inhibitor of the nuclear receptor subfamily 2, group C, member 2 signaling pathway, juxtaposed with another zinc finger gene 1 (JAZF1) has been shown to be involved in gluconeogenesis, lipid metabolism, and insulin sensitivity. However, its role in hepatic and chronic low-grade inflammation leading to nonalcoholic fatty liver disease remains unknown. The aim of this study was to examine whether JAZF1 overexpression in vivo or in vitro can protect against (PA)-induced and high-fat diet (HFD)-induced systemic inflammatory responses, and the potential mechanism of this process. JAZF1 overexpression vector was transfected into PA-treated IAR-20 hepatocytes. The mRNA expression levels of proinflammatory cytokines were measured by real-time quantitative PCR, and stress-activated protein kinase activities were measured by immunoblotting. For in vivo studies, JAZF1 transgenic mice were fed an HFD for 12 weeks. Liver tissue was obtained for histological examination, real-time RT-PCR, and western blot analysis. PA significantly increased the expression levels of tumor necrosis factor-α, monocyte chemotactic protein-1 and interleukin-8 mRNA in IAR-20 hepatocytes in a dose-dependent and time-dependent manner. Treatment with JAZF1 or stress-activated protein kinase inhibitors inhibited PA-induced tumor necrosis factor-α, monocyte chemotactic protein-1 and interleukin-8 expression in these cells. In JAZF1-treated cells, the decreased expression of proinflammatory cytokines was accompanied by decreased p38 mitogen-activated protein kinase and c-Jun N-terminal kinase phosphorylation and increased nuclear factor-κB inhibitor-α protein levels, similarly to the role of signaling inhibitors. In vivo, HFD-induced expression of proinflammatory cytokines was markedly attenuated in JAZF1-Tg mice as compared with controls. This attenuation was accompanied by decreased activation of c-Jun N-terminal kinase, p38 mitogen-activated protein kinase, and nuclear factor-κB. These data provide evidence for the important role of JAZF1 in preventing and systemic inflammation-related disease.© 2014 FEBS.

Keyword: lipogenesis

Hepatic TLR4 signaling in obese NAFLD.

Nonalcoholic fatty liver disease occurs frequently in the setting of metabolic syndrome, but the factors leading to nonalcoholic steatohepatitis (NASH) are not fully understood. This study investigated Toll-like receptor 4 (TLR4) signaling in human liver with the goal of delineating whether activation of this pathway segregates those with nonalcoholic fatty liver from those with NASH. Experiments were performed using liver biopsy tissue obtained from class III obese subjects undergoing bariatric surgery, and extended to an immortalized human hepatocyte HepaRG cell line and primary human hepatocytes. The bacterial endotoxin lipopolysaccharide (LPS) and total free fatty levels were significantly increased in plasma of NASH patients. TLR4 mRNA levels were significantly increased in subjects with NASH compared with NAFL as was interferon regulatory factor (IRF) 3 in the myeloid differentiation factor 88-independent signaling pathway. In HepaRG cells, nuclear factor-κB (NF-κB) nuclear translocation and functional activity increased following treatment with the fatty , palmitate, and following exposure to LPS compared with hepatocytes stimulated with a lipogenic treatment that induced de novo . Palmitate and LPS induction of NF-κB activity was partially attenuated by chemical- or small-interfering RNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was upregulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-κB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition. In conclusion, TLR4 expression is upregulated in a large cohort of NASH patients, compared with those with NAFL, and this occurs within the setting of increased LPS and fatty acids.Copyright © 2015 the American Physiological Society.

Keyword: lipogenesis

De novo and cholesterol synthesis in humans with long-standing type 1 diabetes are comparable to non-diabetic individuals.

Synthesis of lipid species, including fatty acids (FA) and cholesterol, can contribute to pathological disease. The purpose of this study was to investigate FA and cholesterol synthesis in individuals with type 1 diabetes, a group at elevated risk for vascular disease, using stable isotope analysis.Individuals with type 1 diabetes (n = 9) and age-, sex-, and BMI-matched non-diabetic subjects (n = 9) were recruited. On testing day, meals were provided to standardize food intake and elicit typical feeding responses. Blood samples were analyzed at fasting (0 and 24 h) and postprandial (2, 4, 6, and 8 hours after breakfast) time points. FA was isolated from VLDL to estimate hepatic FA synthesis, whereas free cholesterol (FC) and cholesteryl ester (CE) was isolated from plasma and VLDL to estimate whole-body and hepatic cholesterol synthesis, respectively. Lipid synthesis was measured using deuterium incorporation and isotope ratio mass spectrometry.Fasting total hepatic (3.91 ± 0.90% vs. 5.30 ± 1.22%; P = 0.41) was not significantly different between diabetic and control groups, respectively, nor was synthesis of myristic (28.60 ± 4.90% vs. 26.66 ± 4.57%; P = 0.76), (12.52 ± 2.75% vs. 13.71 ± 2.64%; P = 0.65), palmitoleic (3.86 ± 0.91% vs. 4.80 ± 1.22%; P = 0.65), stearic (5.55 ± 1.04% vs. 6.96 ± 0.97%; P = 0.29), and oleic (1.45 ± 0.28% vs. 2.10 ± 0.51%; P = 0.21). Postprandial was also not different between groups (P = 0.38). Similarly, fasting synthesis of whole-body FC (8.2 ± 1.3% vs. 7.3 ± 0.8%/day; P = 0.88) and CE (1.9 ± 0.4% vs. 2.0 ± 0.3%/day; P = 0.96) and hepatic FC (8.2 ± 2.0% vs. 8.1 ± 0.8%/day; P = 0.72) was not significantly different between diabetic and control subjects.Despite long-standing disease, and cholesterol synthesis was not different in individuals with type 1 diabetes compared to healthy non-diabetic humans.

Keyword: lipogenesis

Levels of ester of hydroxystearic (PAHSA) are reduced in the breast milk of obese mothers.

To achieve optimal development of a newborn, breastfeeding is extensively recommended, but little is known about the role of non-nutritive bioactive milk components. We aimed to characterize the fatty esters of hydroxy fatty acids (FAHFAs), namely hydroxystearic acids (PAHSAs)-endogenous lipids with anti-inflammatory and anti-diabetic properties, in human breast milk. Breast milk samples from 30 lean (BMI=19-23) and 23 obese (BMI>30) women were collected 72h postpartum. Adipose tissue and milk samples were harvested from C57BL/6J mice. FAHFA lipid profiles were measured using reverse phase and chiral liquid chromatography-mass spectrometry method. PAHSA regioisomers as well as other FAHFAs were present in both human and murine milk. Unexpectedly, the levels of 5-PAHSA were higher relative to other regioisomers. The separation of both regioisomers and enantiomers of PAHSAs revealed that both R- and S-enantiomers were present in the biological samples, and that the majority of the 5-PAHSA signal is of R configuration. Total PAHSA levels were positively associated with weight gain during pregnancy, and 5-PAHSA as well as total PAHSA levels were significantly lower in the milk of the obese compared to the lean mothers. Our results document for the first time the presence of lipid mediators from the FAHFA family in breast milk, while giving an insight into the stereochemistry of PAHSAs. They also indicate the negative effect of obesity on 5-PAHSA levels. Future studies will be needed to explore the role and mechanism of action of FAHFAs in breast milk.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

PRMT3 regulates hepatic through direct interaction with LXRα.

Arginine methylation is responsible for diverse biological functions and is mediated by protein arginine methyltransferases (PRMTs). Nonalcoholic fatty liver disease (NAFLD) is accompanied by excessive hepatic via liver X receptor α (LXRα). Thus we examined the pathophysiological role of PRMTs in NAFLD and their relationship with LXRα. In this study, (PA) treatment increased PRMT3, which is correlated with the elevation of hepatic lipogenic proteins. The expression of lipogenic proteins was increased by PRMT3 overexpression, but decreased by PRMT3 silencing and use of the PRMT3 knockout (KO) mouse embryonic fibroblast cell line. PRMT3 also increased the transcriptional activity of LXRα by directly binding with LXRα in a methylation-independent manner. In addition, PA treatment translocated PRMT3 to the nucleus. In animal models, a high-fat diet increased the LXRα and PRMT3 expressions and binding, which was not observed in LXRα KO mice. Furthermore, increased PRMT3 expression and its binding with LXRα were observed in NAFLD patients. Taken together, LXRα and PRMT3 expression was increased in cellular and mouse models of NAFLD and human patients, and PRMT3 translocated into the nucleus bound with LXRα as a transcriptional cofactor, which induced . In conclusion, PRMT3 translocation by PA is coupled to the binding of LXRα, which is responsible for the onset of fatty liver.© 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

Keyword: lipogenesis

NLRP3 inflammasome activation in mesenchymal stem cells inhibits osteogenic differentiation and enhances adipogenic differentiation.

Osteoporosis is one of the most common skeletal disease featured by osteopenia and adipose accumulation in bone tissue. NLRP3 inflammasome activation is an essential player in aging-related chronic diseases like osteoporosis, particularly due to the causal caspase-1 activation and its correlation to adipose accumulation in bone tissue. Moreover, the expression of anti-aging/senescence SIRT1 was reported to decline along with aging. As the major cellular contributor of bone formation, mesenchymal stem cells (MSCs) are multipotent stem cells processing mutually exclusive differentiatability toward osteocytes or adipocytes. Therefore, we hypothesized that NLRP3 inflammasome activation promotes and repress osteogenesis in MSCs via inhibiting SIRT1 expression. We activated NLRP3 inflammasome in human MSCs via lipopolysaccharide and (LPS/PA) treatment for self-renewal maintenance, adipogenic differentiation or osteogenic differentiation. LPS/PA treatment significantly increased NLRP3 expression, decreased SIRT1 expression and promoted caspase-1 activity in MSCs. LPS/PA treatment also boosted of MSCs and suppressed osteogenesis. Moreover, inhibition of caspase-1 activity repressed adipogenic differentiation and partially improved osteogenic differentiation of MSCs with LPS/PA treatment. Our study demonstrated the pivotal roles of NLRP3 inflammasome and downstream mediator caspase-1 for the progress of osteo-differentiation MSCs, and offered novel therapeutic target of treatment for osteoporosis.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Macrophage Stimulating Protein Enhances Hepatic Inflammation in a NASH Model.

Non-alcoholic steatohepatitis (NASH) is a common liver disease characterized by hepatic lipid accumulation (steatosis) and inflammation. Currently, therapeutic options are poor and the long-term burden to society is constantly increasing. Previously, macrophage stimulating protein (MSP)-a serum protein mainly secreted by liver-was shown to inhibit oxidized low-density lipoprotein (OxLDL)/lipopolysaccharides (LPS)-induced inflammation in mouse macrophages. Additionally, MSP could reduce (PA)-induced lipid accumulation and in the HepG2 cell line. Altogether, these data suggest MSP as a suppressor for metabolic inflammation. However, so far the potential of MSP to be used as a treatment for NASH was not investigated. We hypothesized that MSP reduces lipid accumulation and hepatic inflammation. To investigate the effects of MSP in the early stage of NASH, low-density lipoprotein receptor (Ldlr-/-) mice were fed either a regular chow or a high fat, high cholesterol (HFC) diet for 7 days. Recombinant MSP or saline (control) was administrated to the mice by utilizing subcutaneously-implanted osmotic mini-pumps for the last 4 days. As expected, mice fed an HFC diet showed increased plasma and hepatic lipid accumulation, as well as enhanced hepatic inflammation, compared with chow-fed controls. Upon MSP administration, the rise in cholesterol and triglyceride levels after an HFC diet remained unaltered. Surprisingly, while hepatic macrophage and neutrophil infiltration was similar between the groups, MSP-treated mice showed increased gene expression of pro-inflammatory and pro-apoptotic mediators in the liver, compared with saline-treated controls. Contrary to our expectations, MSP did not ameliorate NASH. Observed changes in inflammatory gene expression suggest that further research is needed to clarify the long-term effects of MSP.

Keyword: lipogenesis

Heat shock protein 70 promotes in HepG2 cells.

The increasing prevalence of non-alcoholic fatty liver disease (NAFLD) has followed the international rise in obesity rates. Multiple mechanisms are involved in NAFLD, including endoplasmic reticulum stress and oxidative stress. Heat shock protein 70 (HSP70), which is abundant in most organisms, is sensitive to stress. However, the role of HSP70 in NAFLD has not been investigated. Here, we investigated the possible role of HSP70 in lipid synthesis.C57BL/6 mice were fed a high-fat diet, and HepG2 cells were treated with 0.5\xa0mM (PA). HSP70 expression was detected by qPCR, Western blot and immunohistochemistry. Total cholesterol (TC) and triglyceride (TG) levels were detected by enzyme-linked immunosorbent assay (ELISA). After Hsp70 overexpression and knockdown, TC and TG levels and FAS, SCD, and ACC expression were detected.HSP70 expression was significantly increased in the livers of obese mice. In vitro, HSP70 expression was markedly induced by PA in HepG2 cells. Notably, HSP70 overexpression in HepG2 cells enhanced TC and TG synthesis, in parallel with the upregulation of lipogenic genes, including FAS, SCD and ACC. By contrast, HSP70 knockdown decreased the levels of cellular lipids and the expression of FAS, SCD, and ACC in HepG2 cells. Together, our results suggest that HSP70 may promote in HepG2 cells.Heat shock protein 70 promotes in HepG2 cells.

Keyword: lipogenesis

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy metabolism. However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress . In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: lipogenesis

Soraphen A, an inhibitor of acetyl CoA carboxylase activity, interferes with fatty elongation.

Acetyl CoA carboxylase (ACC1 and ACC2) generates malonyl CoA, a substrate for de novo (DNL) and an inhibitor of mitochondrial fatty β-oxidation (FAO). Malonyl CoA is also a substrate for microsomal fatty elongation, an important pathway for saturated (SFA), mono- (MUFA) and polyunsaturated fatty (PUFA) synthesis. Despite the interest in ACC as a target for obesity and cancer therapy, little attention has been given to the role ACC plays in long chain fatty synthesis. This report examines the effect of pharmacological inhibition of ACC on DNL and palmitate (16:0) and linoleate (18:2, n-6) metabolism in HepG2 and LnCap cells. The ACC inhibitor, soraphen A, lowers cellular malonyl CoA, attenuates DNL and the formation of fatty elongation products derived from exogenous fatty acids, i.e., 16:0 and 18:2, n-6; IC(50)∼5nM. Elevated expression of fatty elongases (Elovl5, Elovl6) or desaturases (FADS1, FADS2) failed to override the soraphen A effect on SFA, MUFA or PUFA synthesis. Inhibition of fatty elongation leads to the accumulation of 16- and 18-carbon unsaturated fatty acids derived from 16:0 and 18:2, n-6, respectively. Pharmacological inhibition of ACC activity will not only attenuate DNL and induce FAO, but will also attenuate the synthesis of very long chain saturated, mono- and polyunsaturated fatty acids.Copyright © 2010 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Peroxisome proliferator-activated receptor-gamma protects ERBB2-positive breast cancer cells from palmitate toxicity.

Accumulation of fatty acids and neutral lipids in nonadipose tissues is cytotoxic. We recently showed that ERBB2-positive breast cancer cells produce significantly high amounts of fats, because of overexpression of the peroxisome proliferator-activated receptor (PPAR)gamma-binding protein and the nuclear receptor NR1D1 (nuclear receptor subfamily 1, group D, member 1; Rev-erbalpha). These genes upregulate de novo fatty synthesis, which is a critical pathway for the energy production and survival of these cells. NR1D1 and PPARgamma-binding protein are functionally related to PPARgamma, a well established positive regulator of and lipid storage.The effects of GW9662 and exogenously added palmitate on breast cells (BT474, MDA-MB-361, MCF-7, and human mammary epithelial cells) in monolayer culture were assessed. Mass spectrometric quantitation of fatty acids and fluorescence-based high content microscopy assays of cell growth, apoptosis, triglyceride storage and reactive oxygen species production were used.ERBB2-positive breast cancer cells are more sensitive to inhibition of PPARgamma activity by the antagonist GW9662. PPARgamma inhibition results in increased levels of total fats in the cells, mostly because of increased amounts of and stearic unsaturated acids. Administration of exogenous palmitate is lethal to ERBB2-positive but not to ERBB2-negative cells. GW9662 exacerbates the effects of palmitate addition on BT474 and MDA-MB-361 cells, but it has no significant effect on MCF-7 and human mammary epithelial cells. Palmitate administration results in a fivefold to tenfold greater increase in fat stores in ERBB2-negative cells compared with ERBB2-positive cells, which suggests that the ERBB2-positive cells have maximized their ability to store fats and that additional palmitate is toxic to these cells. Both PPARgamma inhibition and palmitate administration result in increased reactive oxygen species production in BT474 cells. The cell death that results from this treatment can be counteracted by the antioxidant N-acetyl cysteine.Our findings indicate that PPARgamma activity enables ERBB2-positive breast cancer cells, which produce high levels of fat, to convert fatty acids to triglycerides, allowing these cells to avert the cell death that results from lipotoxicity. Endogenous palmitate toxicity represents a genetically based property of ERBB2-positive breast cancer that can be exploited for therapeutic intervention.

Keyword: lipogenesis

PAM, OLA, and LNA are Differentially Taken Up and Trafficked Via Different Metabolic Pathways in Porcine Adipocytes.

Dietary fatty acids have different effects on fat deposition in pigs. To clarify the underlying mechanisms of this difference, we compared the metabolism of (PAM, saturated), oleic (OLA, monounsaturated) and linoleic (LNA, polyunsaturated) in porcine adipocytes treated with 100\xa0μM PAM, OLA or LNA. We observed that the adipocytes incubated with LNA accumulated more lipids compared with those treated with PAM and OLA. We then probed the metabolism of these fatty acids in porcine adipocytes by using isotope-labelled fatty acids. The results showed that 42% of the [1-C] LNA, 34% of the [1-C] PAM and 28% of the [1-C] OLA were recovered in the cellular lipids. The gene expression analyses showed that LNA significantly increased the expression of - and oxidation-related genes including PPARγ, C/EBPα, ap2 and NRF1. In addition, the cells incubated with LNA showed a decreased Ser phosphorylation in PPARγ compared to those incubated with PAM and OLA. Furthermore, when PPARγ Ser phosphorylation was inhibited, no significant difference in the triacylglycerol contents in the adipocytes was observed. These results showed the dietary fatty acids had different metabolism pathways in porcine adipocytes, and LNA significantly promoted lipid accumulation, probably by regulating PPARγ phosphorylation in adipocytes.

Keyword: lipogenesis

Adipogenic/lipogenic gene expression and fatty composition in chuck, loin, and round muscles in response to grain feeding of Yanbian Yellow cattle.

We hypothesized that gene expression and fatty composition would differ among different muscle depots and over time on a grain-based finishing diet. Additionally, we hypothesized that the concentration of SFA would decrease and the concentration of MUFA would increase proportionately with increases in percent intramuscular lipid (%IML). Ten Yanbian Yellow cattle steers (approximately 8 mo of age) were fed a corn-based diet in which the proportion of corn in the ration was increased at 4-mo intervals. Muscle samples were taken by biopsy from the chuck (trapezius), loin (longissimus dorsi), and round (biceps femoris) muscles at 12, 16, 20, 24, and 28 mo of age. The %IML increased from 12 to 28 mo of age, especially between 24 and 28 mo of age, with loin > round > chuck (age × muscle interaction P < 0.001). The percentage (g/100 g total fatty acids) of oleic (18:1n-9), linoleic (18:2n-6), and α-linolenic (18:3n-3), and the MUFA:SFA ratio increased with age, whereas (16:0) and stearic (18:0) decreased with age in all muscles (age effect P < 0.001). The expression of sterol regulatory element binding protein (SREBP1), adipose tissue fatty - binding protein (FABP4), stearoyl-CoA desaturase (SCD), acetyl-CoA carboxylase (ACC1), and lipoprotein lipase (LPL) increased, whereas the expression of peroxisome proliferator-activated receptor gamma (PPARγ) and fatty synthase (FASN) decreased with age. Expression of PPARγ, FABP4, SREBP1, SCD, FASN, ACC1, and LPL was greater in the loin than in the chuck or round (age × muscle interaction P < 0.001), although the MUFA:SFA ratio was greater in the chuck than in the loin or round (muscle effect P < 0.001). In conclusion, adipogenic gene expression was greater in the loin than in the chuck or round muscles, consistent with the greater %IML of the loin. However, the greater SCD gene expression in the loin did not result in a greater amount of MUFA in the loin, relative to the chuck and round.

Keyword: lipogenesis

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: lipogenesis

Synthesis of specific fatty acids contributes to VLDL-triacylglycerol composition in humans with and without type 2 diabetes.

It is recommended that patients with diabetes reduce their intake of saturated fat and increase their intake of monounsaturated fat or carbohydrate. However, high-carbohydrate diets may result in higher saturated fatty acids in VLDL-triacylglycerol. This is attributed to de novo , although synthesis of specific fatty acids is rarely measured. The objective of this study was to examine the contribution of de novo fatty synthesis to VLDL-triacylglycerol composition. It was hypothesised that levels of total and de novo synthesised fatty acids would increase with increased carbohydrate intake in diabetic participants.Seven individuals with type 2 diabetes mellitus and seven matched non-diabetic controls consumed two diets differing in fat energy (lower fat <25%, higher fat >35%) for 3 days in a randomised crossover design. Blood samples were drawn before and 24 h after the ingestion of (2)H-labelled water.In the control participants, the higher-fat diet resulted in a 40% reduction in VLDL-triacylglycerol fatty acids because of decreases in myristic, , palmitoleic and linoleic acids, but the opposite trend occurred in participants with diabetes. The lower-fat diet increased the fractional synthesis rate by 35% and 25% in the control and diabetes participants, respectively (range: 0-33%). Palmitate accounted for 71% of fatty acids synthesised (range: 44-84% total de novo synthesised fatty acids).(2)H incorporation was used for the first time in humans showing variability in the synthesis rate of specific fatty acids, even . A lower-fat diet stimulated saturated fatty synthesis at high rates, but no net stimulation of synthesis of any fatty occurred in the diabetes group. The implications of this finding for our understanding of lipid metabolism in diabetes require further investigation.

Keyword: lipogenesis

Metabolic Fate of Branched-Chain Amino Acids During , in Adipocytes From Obese Mice and C2C12 Myotubes.

Branched-chain amino (BCAA) catabolism is regulated by the branched-chain aminotransferase (BCAT2) and the branched-chain α-keto dehydrogenase complex (BCKDH). BCAT2 and BCKDH expression and activity are modified during and altered in adipose tissues of mice with genetic or diet-induced obesity. However, little is known about how these modifications and alterations affect the intracellular metabolic fate of BCAAs during , in adipocytes from mice fed a control or high-fat diet or in C2C12 myotubes. Here, we demonstrate that BCAAs are mainly incorporated into proteins during the early stages of adipocyte differentiation. However, they are oxidized and incorporated into lipids during the late days of differentiation. Conversely, 92% and 97% of BCAA were oxidized, 1.6% and 6% were used for protein synthesis and 1.2% and 1.5% were incorporated into lipids in adipocytes from epididymal and subcutaneous adipose tissue, respectively. All three pathways were decreased in adipocytes from mice fed a high-fat diet. In C2C12 myotubes, leucine is mainly used for protein synthesis and palmitate is incorporated into lipids. Interestingly, leucine decreased both palmitate oxidation and its incorporation to lipids and proteins; and palmitate increased leucine oxidation and decreased its incorporation to lipids and proteins in a dose-dependent manner. These results demonstrate that BCAA metabolic fate differs between the early and late stages of adipocyte differentiation and in adipocytes from mice fed a control or high-fat diet; and that leucine affects the metabolic fate of palmitate and vice versa in C2C12 myotubes. J. Cell. Biochem. 118: 808-818, 2017.© 2016 Wiley Periodicals, Inc.

Keyword: lipogenesis

Insulin-like growth factor binding protein 7 accelerates hepatic steatosis and insulin resistance in non-alcoholic fatty liver disease.

An association between increased insulin-like growth factor binding protein-7 (IGFBP7) expression and insulin resistance in metabolic diseases has been reported. However, the role and molecular mechanism of IGFBP-7 in non-alcoholic fatty liver disease (NAFLD) remains largely unknown. Therefore, the potential function of IGFBP7 in the pathological progression of NAFLD was explored in this investigation. For in vivo experiments, an animal model of NAFLD was established in C57BL/6 mice by feeding a high-fat diet (HFD), and IGFBP7 was knocked down by injecting adeno-associated adenovirus (AAV)-mediated short-hairpin (sh)-IGFBP7 into the liver. We found that AAV-sh-IGFBP7 treatment significantly alleviated hepatocyte injury and inhibited hepatic lipid accumulation by reducing -associated gene expression. Furthermore, downregulation of IGFBP7 markedly ameliorated IR and restored impaired insulin signalling by elevating the phosphorylation levels of IRS-1, Akt and GSK3β in HFD-treated mice. Similar results were also confirmed by an in vitro study in a (PA)-stimulated HepG2 cell model. In conclusion, our study demonstrates that IGFBP7 contributes to hepatic steatosis and insulin resistance in NAFLD development, which might serve as a novel therapeutic agent for the treatment of NAFLD.© 2019 John Wiley & Sons Australia, Ltd.

Keyword: lipogenesis

Fatty Composition of Plasma Phosphatidylcholine Determines Body Fat Parameters in Subjects with Metabolic Syndrome-Related Traits.

This study examines the associations of fatty acids (FAs) in plasma phosphatidylcholine (PC) with the anthropometrical and biochemical characteristic of patients with metabolic syndrome (MetS)-related traits.We analyzed the FA profiles of PC in 300 persons with MetS-related traits (152\u2009M/148F, mean age 46.9\u2009±\u20099.0 years) and in 70 healthy controls of the same age using a balanced men/women ratio and gas-liquid chromatography. Multivariate linear regression analysis was performed to determine the coefficients of determination (R) using FA proportions of the mentioned proband characteristics.The FA composition of PC in patients with MetS traits was only associated with waist circumference (R\u2009=\u20090.27), waist-to-hip ratio (WHR; R\u2009=\u20090.41), body fat percentage (R\u2009=\u20090.62), and fat mass (R\u2009=\u20090.29). Positive associations were found for dihomo-γ-linolenic (DGLA), , stearic (SA), α-linolenic (ALA), and eicosapentaenoic acids, whereas negative associations were found for linoleic (LA), oleic, and docosapentaenoic acids. Palmitoleic (POA) was positively associated with waist circumference but negatively with fat percentage. In controls, significant associations were found for waist circumference (R\u2009=\u20090.51), WHR (R\u2009=\u20090.53), body fat percentage (R\u2009=\u20090.60), and fat mass (R\u2009=\u20090.34). DGLA and saturated FA (SFA) were positively associated, whereas docosahexaenoic, adrenic, and cis-vaccenic acids were negatively associated. The study group differed from controls as follows: lower concentrations of LA and total n-6 FA, higher indices of delta-9-desaturase and delta-6 desaturase activity and higher proportions of POA, SA, ALA, DGLA, and SFA.We found significant associations (R >0.25) of FA in plasma PC with adiposity in middle-aged persons with MetS-related traits, but not with metabolic indices.

Keyword: lipogenesis

Fatty biosynthesis and lipogenic enzyme activities in subcutaneous adipose tissue of feedlot steers fed supplementary palm oil or soybean oil.

We hypothesized that supplementing finishing diets with palm oil would promote adipocyte differentiation in subcutaneous adipose tissue of feedlot steers, and that soybean oil supplementation would depress adipocyte differentiation. Twenty-eight Angus steers were assigned randomly to 3 groups of 9 or 10 steers and fed a basal diet without additional fat (control), with 3% palm oil (rich in ), or with 3% soybean oil (rich in polyunsaturated fatty acids), for 10 wk, top-dressed daily. Palm oil had no effect (P > 0.05) on ADG, food intake, or G:F, whereas soybean oil depressed ADG (P = 0.02), food intake (P = 0.04), and G:F (P = 0.05). Marbling scores tended (P = 0.09) to be greater in palm oil-fed steers (Modest(09)) than in soybean oil-fed steers (Small(55)). Subcutaneous adipocyte mean volume was greater in palm oil-fed steers (515.9 pL) than in soybean-supplemented cattle (395.6 pL; P = 0.01). Similarly, glucose and acetate incorporation into total lipids in vitro was greater in subcutaneous adipose tissue of palm oil-fed steers (119.9 and 242.8 nmol·3h(-1)·10(5) cells, respectively) than adipose tissue of soybean oil-fed steers in (48.9 and 95.8 nmol·3h(-1)·10(5) cells, respectively). Glucose-6-phosphate dehydrogenase and NADP-malate dehydrogenase activities were greater (P ≤ 0.05) in subcutaneous adipose tissue of palm oil-fed steers than in adipose tissue of control steers. Palm oil did not increase or decrease oleic in subcutaneous adipose tissue or LM, but decreased (P ≤ 0.05) myristoleic, palmitoleic, and cis-vaccenic in adipose tissue, indicating a depression in stearoyl-coenzyme A desaturase activity. Soybean oil increased the proportion of α-linolenic in adipose tissue and muscle and increased linoleic and 18:1trans-10 in muscle. We conclude that palm oil supplementation promoted lipid synthesis in adipose tissue without depressing feed efficiency or increasing the content of beef.

Keyword: lipogenesis

Mechanisms of lipid mobilization towards egg formation in broiler breeder hens using stable isotopes.

The contribution of dietary, mobilized, or newly synthesized fatty acids in yolk formation at different periods of egg production was determined. In an initial experiment, a single dose of C-linoleic was administered to pullets at the onset of egg production and their presence in follicles determined over the subsequent 10 days. In a second experiment, pullets were fed a daily 15\xa0mg dose of U-C-glucose beginning 2 wk prior to sexual maturity through the end of the experimental period. A 50\xa0mg meal of U-C-linoleic was orally administered approximately 10\xa0d prior to sexual maturity (defined as first egg) representing body linoleic . Upon each hen\'s first egg, each bird received a 25\xa0mg meal of D-linoleic representing dietary linoleic . All eggs were collected for the next 10 days. The incorporation of labeled linoleic and in egg yolk was then determined using GC-MS. This process was repeated at peak production and at 45 wk of age. At sexual maturity, the deposition of labeled in the yolk was higher compared with its deposition at peak production and 45 wk of age. The deposition of both C- and D-linoleic increased with hen age. These results suggest that dietary and tissue linoleic is utilized to a greater extent in older hens and that (synthesis of ) plays a larger role at sexual maturity in the young hen.© 2016 Poultry Science Association Inc.

Keyword: lipogenesis

Prepartum fatty supplementation in sheep. II. Supplementation of eicosapentaenoic and docosahexaenoic during late gestation alters the fatty profile of plasma, colostrum, milk and adipose tissue, and increases lipogenic gene expression of adipose tissue.

The objectives of this study were as follows: 1) to establish whether feeding a source of eicosapentaenoic (EPA) and docosahexaenoic (DHA) to ewes during late gestation changes the fatty profile of colostrum, milk, ewe adipose tissue, and plasma and subsequently lamb plasma and red blood cells (RBC), and 2) to investigate the effects of EPA and DHA on mRNA expression in ewe adipose tissue. Eighty-four gestating ewes (28 pens, three per pen) were blocked by lambing day and assigned to a diet with an addition of fat at 0.39% of the DM during the last 50 d of gestation using Ca salts of a palm fatty distillate (PFAD) high in and oleic acids or EPA + DHA. Blood samples were taken from ewes on days 20, 1 (parturition), and 30 and from lambs on days 1 and 30 for plasma fatty analysis. Fatty analysis of lamb RBC was performed on day 1. Colostrum samples were taken at lambing and milk samples on day 30 for fatty analysis. Subcutaneous adipose tissue biopsies were taken from one ewe per pen on day 20 for fatty analysis and gene expression analysis of 27 genes. Treatment × day interactions (P < 0.10) were observed for several isomers of C18:1, with concentrations that were greater in plasma of EPA + DHA ewes on day 20, but were not different on day 1 or 30. Plasma concentrations of EPA tended to be greater (P = 0.07), whereas DHA was greater (P < 0.001) in EPA + DHA ewes compared with PFAD ewes. There was no difference in EPA or DHA in adipose tissue with EPA + DHA vs. PFAD supplementation (P > 0.10). Concentrations of fatty acids with 6 to 10 carbons were significantly increased (P < 0.05) in colostrum and milk of EPA + DHA ewes. There was a treatment × day interaction with EPA + DHA ewes yielding greater EPA (P = 0.03) and DHA (P = 0.04) concentrations than PFAD in colostrum, but not in milk. Treatment × day interactions (P < 0.05) were observed for several C18:1 isomers with concentrations that were greater in EPA + DHA ewe colostrum, but were not different between treatments in milk. In lamb plasma and RBC, EPA and DHA were not different between treatments (P > 0.10). The expression of fatty synthase and leptin was significantly increased (P < 0.05), whereas the expression of diacylglycerol acyltransferase 2 tended to be increased (P = 0.08) by supplementation of EPA + DHA vs. PFAD. These results suggest that supplementation with EPA and DHA to ewes during late gestation alters the fatty profile of plasma, colostrum, and milk and may increase .

Keyword: lipogenesis

Adipocytes differentiated in vitro from rat mesenchymal stem cells lack essential free fatty acids compared to adult adipocytes.

Adult bone marrow mesenchymal stem cells (BMSCs) can be differentiated in vitro to become adipocyte-like cells with lipid vacuoles, similar to adipocytes derived from adult adipose tissue. Little is known regarding the composition of free fatty acids (FFAs) of the in vitro-differentiated adipocytes, or whether it resembles that of native adult adipocytes. We used gas chromatography-mass spectrometry to identify FFA species in BMSC-derived adipocytes and compared them with FFAs found in adipocytes derived from adult adipose tissue. We found that adult adipocytes contained significant percentages of saturated and monounsaturated FFAs, including (C16:0), stearic (C18:0), and oleic (C18:1); some polyunsaturated FFAs, such as linoleic (C18:2), a small percentage of arachidonic (C20:4), and very little linolenic (C18:3). In comparison, 80%-90% confluent BMSCs contained comparable percentages of and oleic acids, significantly more arachidonic and stearic acids, very little linoleic , and no linolenic . After differentiation, compared with adult adipocytes, BMSC-derived adipocytes contained a comparable percentage of , more stearic and arachidonic acids, less oleic , almost no linoleic , and no detectable linolenic . This composition was quite similar to that of undifferentiated BMSCs. The differentiation medium contained only and stearic acids, with traces of oleic ; it did not contain the essential polyunsaturated fatty acids. Thus, the composition of FFAs in BMSC-derived adipocytes was altered compared with adult adipocytes. BMSC-derived adipocytes had an altered composition of saturated and monounsaturated FFAs and lacked essential FFAs that may directly affect signaling related to their lipolysis/ functions.

Keyword: lipogenesis

Lipid body biogenesis and the role of microtubules in lipid synthesis in Ornithogalum umbellatum lipotubuloids.

Lipid bodies present in lipotubuloids of Ornithogalum umbellatum ovary epidermis take the form of a lens between leaflets of ER (endoplasmic reticulum) membrane filled with a highly osmiophilic substance. The two enzymes, DGAT1 [DAG (diacylglycerol) acyltransferase 1] and DGAT2 (DAG acyltransferase 2), involved in this process are synthesized on rough ER and localized in the ER near a monolayer surrounding entities like lipid bodies. After reaching the appropriate size, newly formed lipid bodies transform into mature spherical lipid bodies filled with less osmiophilic content. They appear to be surrounded by a half-unit membrane, with numerous microtubules running adjacently in different directions. The ER, no longer continuous with lipid bodies, makes contact with them through microtubules. At this stage, lipid synthesis takes place at the periphery of lipid bodies. This presumption, and a hypothesis that microtubules are involved in lipid synthesis delivering necessary components to lipid bodies, is based on strong arguments: (i) silver grains first appear over microtubules after a short [3H] incubation and before they are observed over lipid bodies; (ii) blockade of [3H] incorporation into lipotubuloids by propyzamide, an inhibitor of microtubule function; and (iii) the presence of gold grains above the microtubules after DGAT1 and DGAT2 reactions, as also near microtubules after an immunogold method that identifies phospholipase D1.

Keyword: lipogenesis

induces central leptin resistance and impairs hepatic glucose and lipid metabolism in male mice.

The consumption of diets rich in saturated fat largely contributes to the development of obesity in modern societies. A diet high in saturated fats can induce inflammation and impair leptin signaling in the hypothalamus. However, the role of saturated fatty acids on hypothalamic leptin signaling, and hepatic glucose and lipid metabolism remains largely undiscovered. In this study, we investigated the effects of intracerebroventricular (icv) administration of a saturated fatty , (PA, C16:0), on central leptin sensitivity, hypothalamic leptin signaling, inflammatory molecules and hepatic energy metabolism in C57BL/6J male mice. We found that the icv administration of PA led to central leptin resistance, evidenced by the inhibition of central leptin\'s suppression of food intake. Central leptin resistance was concomitant with impaired hypothalamic leptin signaling (JAK2-STAT3, PKB/Akt-FOXO1) and a pro-inflammatory response (TNF-α, IL1-β, IL-6 and pIκBa) in the mediobasal hypothalamus and paraventricular hypothalamic nuclei. Furthermore, the pre-administration of icv PA blunted the effect of leptin-induced decreases in mRNA expression related to gluconeogenesis (G6Pase and PEPCK), glucose transportation (GLUT2) and (FAS and SCD1) in the liver of mice. Therefore, elevated central PA concentrations can induce pro-inflammatory responses and leptin resistance, which are associated with disorders of energy homeostasis in the liver as a result of diet-induced obesity.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Suppression of via Reactive Oxygen Species-AMPK Signaling for Treating Malignant and Proliferative Diseases.

Systemic diseases often have common characteristics. The aim of this study was to investigate the feasibility of targeting common pathological metabolism to inhibit the progression of malignant and proliferative diseases.Gefitinib-resistant (G-R) nonsmall-cell lung cancer (NSCLC) and rheumatoid arthritis (RA) were studied as conditions representative of malignant and proliferative diseases, respectively. Strong lipogenic activity and high expression of sterol regulatory element-binding protein 1 (SREBP1) were found in both G-R NSCLC cells and synovial fibroblasts from RA patients (RASFs). Berberine (BBR), an effective suppressor of SREBP1 and regulated through reactive oxygen species (ROS)/AMPK pathway, selectively inhibited the growth of G-R NSCLC cells and RASFs but not that of normal cells. It effectively caused mitochondrial dysfunction, activated ROS/AMPK pathway, and finally suppressed cellular and cell proliferation. Addition of ROS blocker, AMPK inhibitor, and significantly reduced the effect of BBR. In an in vivo study, treatment of BBR led to significant inhibition of mouse tumor xenograft growth and remarkably slowed down the development of adjuvant-induced arthritis in rats. Innovation and Conclusion: Targeting ROS/AMPK/ signaling pathway selectively inhibited the growth of G-R NSCLC cells and the progress of RASFs in vitro and in vivo, which provides a new avenue for treating malignancies and proliferative diseases. Antioxid. Redox Signal. 28, 339-357.

Keyword: lipogenesis

Factors associated with the content of mammary-synthesized fatty acids in milk fat: A meta-analysis.

Consumption of specific fatty acids (FA) that are synthesized in the mammary gland, namely de novo FA, has implications for human health. The objective of the present meta-analysis was to study the associations between milk fat content of de novo FA, with (1) diet composition, and (2) milk production and composition. Milk FA data from 96 peer-reviewed studies published between 1990 and 2016 that included 324 treatment means from 83 bovine experiments, 36 treatment means from 12 caprine experiments, and 40 treatment means from 12 ovine experiments were used in this analysis. Individual species models including the fixed effect of experiment were fitted using multiple regression to explain milk content of de novo FA as a function of diet composition and milk production and composition variables. We also evaluated replacing the effect of the experiment by the effect of the experiment nested in the laboratory at which the research had been conducted, and the effect of the laboratory. Butyric content in milk fat was positively but weakly related to dietary ether extract in does and ewes. Lauric, myristic, and contents in milk fat were negatively related to dietary ether extract in does and to a somewhat lesser extent in cows and ewes. The results confirm that the inclusion of lipids in the diet may not only affect the availability of preformed FA but also the profile of FA synthesized de novo in the mammary gland. Most of the variation in all prediction models was explained by the experiment or by the laboratory if the latter was included in the model. The ample variation in analytical methods reported by the different research groups suggests that differences in analytical protocols might explain a substantial proportion of the variation in de novo FA profile. A main conclusion of this study is the potential influence of differences in analytical procedures to explain the variation in de novo FA profile. Standardization of methods of FA analysis to improve reproducibility seems to be an aspect of importance to this area of research.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Absence of adipose triglyceride lipase protects from hepatic endoplasmic reticulum stress in mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by triglyceride (TG) accumulation and endoplasmic reticulum (ER) stress. Because fatty acids (FAs) may trigger ER stress, we hypothesized that the absence of adipose triglyceride lipase (ATGL/PNPLA2)-the main enzyme for intracellular lipolysis, releasing FAs, and closest homolog to adiponutrin (PNPLA3) recently implicated in the pathogenesis of NAFLD-protects against hepatic ER stress. Wild-type (WT) and ATGL knockout (KO) mice were challenged with tunicamycin (TM) to induce ER stress. Serum biochemistry, hepatic TG and FA profiles, liver histology, and gene expression for markers of hepatic lipid metabolism, ER stress, and inflammation were explored. Moreover, cell-culture experiments were performed in Hepa1.6 cells after the knockdown of ATGL before FA and TM treatment. TM increased hepatic TG accumulation in ATGL KO, but not in WT, mice. and β-oxidation were repressed at the gene-expression level (sterol regulatory element-binding transcription factor 1c, fatty synthase, acetyl coenzyme A carboxylase 2, and carnitine palmitoyltransferase 1 alpha) in both WT and ATGL KO mice. Genes for very-low-density lipoprotein (VLDL) synthesis (microsomal triglyceride transfer protein and apolipoprotein B) were down-regulated by TM in WT and even more in ATGL KO mice, which displayed strongly reduced serum VLDL cholesterol levels. Notably, ER stress markers glucose-regulated protein, C/EBP homolog protein, spliced X-box-binding protein, endoplasmic-reticulum-localized DnaJ homolog 4, and inflammatory markers Tnfα and iNos were induced exclusively in TM-treated WT, but not ATGL KO, mice. Total hepatic FA profiling revealed a higher /oleic (PA/OA) ratio in WT mice, compared to ATGL KO mice, at baseline. Phosphoinositide-3-kinase inhibitor-known to be involved in FA-derived ER stress and blocked by OA-was increased in TM-treated WT mice only. In line with this, in vitro OA protected hepatocytes from TM-induced ER stress.Lack of ATGL may protect from hepatic ER stress through alterations in FA composition. ATGL could constitute a new therapeutic strategy to target ER stress in NAFLD.Copyright © 2012 American Association for the Study of Liver Diseases.

Keyword: lipogenesis

1,25-Dihydroxyvitamin D regulates lipid metabolism and glucose utilization in differentiated 3T3-L1 adipocytes.

It is well established that adipose tissue can both store and metabolize vitamin D. The active form of vitamin D, 1,25 dihydroxyvitamin D [1,25(OH)D], regulates adipocyte differentiation and inflammation, highlighting the multifaceted role that vitamin D plays in adipose tissue physiology. However, there is limited evidence regarding vitamin D regulation of mature adipocyte lipid metabolism. We hypothesize that 1,25(OH)D alters lipid and glucose metabolism in differentiated 3T3-L1 adipocytes to reduce triacylglycerol (TAG) accumulation. In this study, 1,25(OH)D (10\u202fnmol/L) stimulated a 21% reduction in TAG accumulation in differentiated 3T3-L1 adipocytes after 4\u202fdays (P\u202f=\u202f.01) despite a significant increase in fatty uptake (P\u202f<\u202f.01). Additionally, 1,25(OH)D stimulated a 2.5-fold increase in CO production from [1-C] (P\u202f<\u202f.01), indicative of an elevated rate of fatty β-oxidation, while stimulating a 9% reduction in de novo fatty synthesis (P\u202f=\u202f.03). Interestingly, d-[U-C]glucose incorporation into fatty acids was reduced by 30% in response to 1,25(OH)D (P\u202f<\u202f.01), indicating a reduced contribution of glucose as a substrate for de novo . Consistent with these findings, mRNA expression of the anaplerotic enzyme pyruvate carboxylase was reduced by 41% (P\u202f<\u202f.01). In summary, 1,25(OH)D stimulated fatty oxidation and reduced TAG accumulation in differentiated adipocytes. Furthermore, 1,25(OH)D reduced glucose utilization as a substrate for fatty synthesis potentially by downregulating pyruvate carboxylase and stimulating glucose disposal as glycerol. Collectively, these 1,25(OH)D-induced changes in lipid metabolism and glucose utilization may contribute to the reduction in TAG accumulation and be protective against excessive fat mass accumulation and associated metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Pleiotropic effects of the goat prolactin receptor genotype on milk fatty composition.

In the lactating mammary gland, prolactin (PRL) stimulates the synthesis of lactose as well as fatty uptake, , and triacylglycerol synthesis. Associations between bovine PRL receptor (PRLR) genotype and fat yield have been reported, which illustrates the role of PRL in conveying lipids toward the udder as well as in stimulating their local synthesis during lactation. Conversely, and to the best of our knowledge, the effects of PRLR genotype on milk fatty content have not been studied so far in any mammalian species. In this study, we sequenced most of the coding region of the caprine PRLR gene in several individuals from the Malagueña and Murciano-Granadina breeds. This approach allowed us to identify 2 long and short mRNA isoforms, produced by alternative splicing, and 4 single-nucleotide polymorphisms (SNPs), namely, c.177T>C, c.1131G>A, c.1201G>A and c.1355C>T. Two of these SNPs are nonsynonymous and involve G401R (c.1201G>A) and T452I (c.1355C>T) amino substitutions in the cytoplasmic domain of PRLR, which plays a fundamental role in signal transduction. Performance of an association analysis with milk composition traits in a Murciano-Granadina goat population revealed highly suggestive effects on palmitoleic content, whereas suggestive effects were detected for other fatty acids, such as and linoleic. These results are consistent with the pleiotropic effects of PRL on mammary gland lipid metabolism and milk composition.Copyright 2010 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

High Fat Diet Upregulates Fatty Oxidation and Ketogenesis via Intervention of PPAR-γ.

Systemic hyperlipidemia and intracellular lipid accumulation induced by chronic high fat diet (HFD) leads to enhanced fatty oxidation (FAO) and ketogenesis. The present study was aimed to determine whether activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) by surplus free fatty acids (FA) in hyperlipidemic condition, has a positive feedback regulation over FAO and ketogenic enzymes controlling lipotoxicity and cardiac apoptosis.8 weeks old C57BL/6 wild type (WT) or PPAR-γ-/- mice were challenged with 16 weeks 60% HFD to induce obesity mediated type 2 diabetes mellitus (T2DM) and diabetic cardiomyopathy. Treatment course was followed by echocardiographic measurements, glycemic and lipid profiling, immunoblot, qPCR and immunohistochemistry (IHC) analysis of PPAR-γ and following mitochondrial metabolic enzymes 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), mitochondrial β- hydroxy butyrate dehydrogenase (BDH1) and pyruvate dehydrogenase kinase isoform 4 (PDK4). In vivo model was translated in vitro, with neonatal rat cardiomyocytes (NRCM) treated with PPAR-γ agonist/antagonist and PPAR-γ overexpression adenovirus in presence of (PA). Apoptosis was determined in vivo from left ventricular heart by TUNEL assay and immunoblot analysis.We found exaggerated circulating ketone bodies production and expressions of the related mitochondrial enzymes HMGCS2, BDH1 and PDK4 in HFD-induced diabetic hearts and in PA-treated NRCM. As a mechanistic approach we found HFD mediated activation of PPAR-γ is associated with the above-mentioned mitochondrial enzymes. HFD-fed PPAR-γ-/-mice display decreased hyperglycemia, hyperlipidemia associated with increased insulin responsiveness as compared to HFD-fed WT mice PPAR-γ-/-HFD mice demonstrated a more robust functional recovery after diabetes induction, as well as significantly reduced myocyte apoptosis and improved cardiac function.PPAR-γ has been described previously to regulate lipid metabolism and . The present study suggests for the first time that increased PPAR-γ expression by HFD is responsible for cardiac dysfunction via upregulation of mitochondrial enzymes HMGCS2, BDH1 and PDK4. Targeting PPAR-γ and its downstream mitochondrial enzymes will provide novel strategies in preventing metabolic and myocardial dysfunction in diabetes mellitus.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: lipogenesis

Free fatty -induced PP2A hyperactivity selectively impairs hepatic insulin action on glucose metabolism.

In type 2 Diabetes (T2D) free fatty acids (FFAs) in plasma are increased and hepatic insulin resistance is "selective", in the sense that the insulin-mediated decrease of glucose production is blunted while insulin\'s effect on stimulating is maintained. We investigated the molecular mechanisms underlying this pathogenic paradox. Primary rat hepatocytes were exposed to palmitate for twenty hours. To establish the physiological relevance of the in vitro findings, we also studied insulin-resistant Zucker Diabetic Fatty (ZDF) rats. While insulin-receptor phosphorylation was unaffected, activation of Akt and inactivation of the downstream targets Glycogen synthase kinase 3α (Gsk3α and Forkhead box O1 (FoxO1) was inhibited in palmitate-exposed cells. Accordingly, dose-response curves for insulin-mediated suppression of the FoxO1-induced gluconeogenic genes and for de novo glucose production were right shifted, and insulin-stimulated glucose oxidation and glycogen synthesis were impaired. In contrast, similar to findings in human T2D, the ability of insulin to induce triglyceride (TG) accumulation and transcription of the enzymes that catalyze de novo and TG assembly was unaffected. Insulin-induction of these genes could, however, be blocked by inhibition of the atypical PKCs (aPKCs). The activity of the Akt-inactivating Protein Phosphatase 2A (PP2A) was increased in the insulin-resistant cells. Furthermore, inhibition of PP2A by specific inhibitors increased insulin-stimulated activation of Akt and phosphorylation of FoxO1 and Gsk3α. Finally, PP2A mRNA levels were increased in liver, muscle and adipose tissue, while PP2A activity was increased in liver and muscle tissue in insulin-resistant ZDF rats. In conclusion, our findings indicate that FFAs may cause a selective impairment of insulin action upon hepatic glucose metabolism by increasing PP2A activity.

Keyword: lipogenesis

Diabetes: Hepatic independent of insulin in type 2 diabetes mellitus--a paradox clarified.

Keyword: lipogenesis

Ruscogenin ameliorates experimental nonalcoholic steatohepatitis via suppressing and inflammatory pathway.

The aim of the study was to investigate the protective effects of ruscogenin, a major steroid sapogenin in Ophiopogon japonicus, on experimental models of nonalcoholic steatohepatitis. HepG2 cells were exposed to 300\u2009μmol/l (PA) for 24\u2009h with the preincubation of ruscogenin for another 24\u2009h. Ruscogenin (10.0\u2009μmol/l) had inhibitory effects on PA-induced triglyceride accumulation and inflammatory markers in HepG2 cells. Male golden hamsters were randomly divided into five groups fed a normal diet, a high-fat diet (HFD), or a HFD supplemented with ruscogenin (0.3, 1.0, or 3.0\u2009mg/kg/day) by gavage once daily for 8 weeks. Ruscogenin alleviated dyslipidemia, liver steatosis, and necroinflammation and reversed plasma markers of metabolic syndrome in HFD-fed hamsters. Hepatic mRNA levels involved in fatty oxidation were increased in ruscogenin-treated HFD-fed hamsters. Conversely, ruscogenin decreased expression of genes involved in hepatic . Gene expression of inflammatory cytokines, chemoattractive mediator, nuclear transcription factor-(NF-) κB, and α-smooth muscle actin were increased in the HFD group, which were attenuated by ruscogenin. Ruscogenin may attenuate HFD-induced steatohepatitis through downregulation of NF-κB-mediated inflammatory responses, reducing hepatic lipogenic gene expression, and upregulating proteins in β-oxidation pathway.

Keyword: lipogenesis

MSP is a negative regulator of inflammation and in ex vivo models of non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (NASH), a metabolic disorder consisting of steatosis and inflammation, is considered the hepatic equivalent of metabolic syndrome and can result in irreversible liver damage. Macrophage-stimulating protein (MSP) is a hepatokine that potentially has a beneficial role in hepatic lipid and glucose metabolism via the activation of AMP-activated protein kinase (AMPK). In the current study, we investigated the regulatory role of MSP in the development of inflammation and lipid metabolism in various NASH models, both in vitro and ex vivo. We observed that MSP treatment activated the AMPK signaling pathway and inhibited lipopolysaccharide (LPS)- and (PA)-induced gene expression of pro-inflammatory cytokines in primary mouse hepatocytes. In addition, MSP treatment resulted in a significant reduction in PA-induced lipid accumulation and inhibited the gene expression of key lipogenic enzymes in HepG2 cells. Upon short hairpin RNA-induced knockdown of RON (the membrane-bound receptor for MSP), the anti-inflammatory and anti-lipogenic effects of MSP were markedly ablated. Finally, to mimic NASH ex vivo, we challenged bone marrow-derived macrophages with oxidized low-density lipoprotein (oxLDL) in combination with LPS. OxLDL+LPS exposure led to a marked inhibition of AMPK activity and a robust increase in inflammation. MSP treatment significantly reversed these effects by restoring AMPK activity and by suppressing pro-inflammatory cytokine gene expression and secretion under this condition. Taken together, these data suggest that MSP is an effective inhibitor of inflammation and lipid accumulation in the stressed liver, thereby indicating that MSP has a key regulatory role in NASH.

Keyword: lipogenesis

The IRE1α-XBP1s pathway promotes insulin-stimulated glucose uptake in adipocytes by increasing PPARγ activity.

The peroxisome proliferator-activated receptor-γ (PPARγ) improves whole-body insulin sensitivity by regulating the adipogenic and metabolic functions of mature adipocytes. We have previously demonstrated that an active splice variant of X-box binding protein 1 (XBP1s) enhances PPARγ expression during . In this study, we investigated the role of XBP1s, particularly with respect to PPARγ, in the mechanisms underlying insulin sensitivity in mature adipocytes. Insulin was able to stimulate XBP1s generation by activating inositol-requiring enzyme 1 (IRE1) α and was also able to increase its transcriptional activity by inducing nuclear translocation. XBP1s also upregulated the levels of phosphorylated IRS1 and AKT, demonstrating a positive feedback regulatory mechanism linking insulin and XBP1s. XBP1s enhanced the expression of fibroblast growth factor 21 and, in turn, increased PPARγ activity, translocation of GLUT4 to the cell surface, and glucose uptake rate in adipocytes. In addition, XBP1s abolished palmitate-induced insulin resistance in adipocytes by increasing adiponectin secretion, repressing the secretion of pro-inflammatory adipokines such as leptin, monocyte chemoattractant protein 1, and tumor necrosis factor α, and decreasing fatty release. These findings provide a novel mechanism by which XBP1s stimulate insulin sensitivity in adipocytes through fibroblast growth factor 21 induction and PPARγ activation.

Keyword: lipogenesis

Role for sterol regulatory element binding protein-1c activation in mediating skeletal muscle insulin resistance via repression of rat insulin receptor substrate-1 transcription.

Sterol regulatory element binding protein-1c (SREBP-1c) is a master regulator of fatty synthase and controls . IRS-1 is the key insulin signalling mediator in skeletal muscle. In the present study, we investigated the role of SREBP-1c in the regulation of IRS-1 in skeletal muscle cells.L6 muscle cells were treated with (PA) or metformin. Adenovirus vectors expressing Srebp-1c (also known as Srebf1) and small interfering RNA (siRNA) against Srebp-1c were transfected into the L6 cells. Protein-DNA interactions were assessed by luciferase reporter analysis, electrophoretic mobility shift assay and chromatin immunoprecipitation assay.We found that both gene and protein expression of SREBP-1c was increased in contrast to IRS-1 expression in PA-treated L6 cells. SREBP-1c overproduction decreased Irs-1 mRNA and IRS-1 protein expression in a dose-dependent manner, and suppressed the resultant insulin signalling, whereas SERBP-1c knockdown by Serbp-1c siRNA blocked the downregulation of IRS-1 induced by PA. Protein-DNA interaction studies demonstrated that SREBP-1c was able to bind to the rat Irs-1 promoter region, thereby repressing its gene transcription. Of particular importance, we found that metformin treatment downregulated Srebp-1c promoter activity, decreased the specific binding of SREBP-1c to Irs-1 promoter and upregulated Irs-1 promoter activity in PA-cultured L6 cells.Our data indicate for the first time that SREBP-1c activation participates in skeletal muscle insulin resistance through a direct effect of suppressing Irs-1 transcription. These findings imply that SREBP-1c could serve as an attractive therapeutic target for insulin resistance.

Keyword: lipogenesis

Plasma fatty acids in de novo lipogenesis pathway are associated with diabetogenic indicators among adults: NHANES 2003-2004.

Insulin regulates fatty acids (FAs) in the blood; conversely, FAs may mediate insulin sensitivity and are potentially modifiable risk factors of the diabetogenic state.The objective of our study was to examine the associations between plasma concentrations of FAs, fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among individuals (n\xa0=\xa01433) in the NHANES (2003-2004).Plasma concentrations of 24 individual FAs were considered individually and in subgroups, per chemical structure. Study participants were categorized in diabetogenic groups: Group 1 (HbA1c ≥6.5% or FPG ≥126 mg/dL), Group 2 (HbA1c 5.7% to <6.5% or FPG 100 to <126 mg/dL), and Group 3 (HbA1c <5.7% and FPG <100 mg/dL). We assessed associations between diabetogenic groups and plasma FAs in multivariate multinomial regressions (with Group 3 as the reference).Overall, 7.0% of study participants were in Group 1; 33.3% were in Group 2. Plasma concentrations of several individual FAs, including even-chain saturated FAs (SFAs; myristic, , stearic acids) and monounsaturated FAs (MUFAs; cis-vaccenic, oleic acids), were respectively associated with greater odds of Groups 1 and 2 status, adjusting for covariates. Higher concentrations of SFA and MUFA subgroups (highest compared with lowest quartile) were associated with increased odds of Group 2 status [SFAs adjusted OR (aOR): 1.51 (95% CI: 1.05, 2.18); MUFAs aOR: 1.78 (95% CI: 1.11, 2.85)]. Higher eicosapentaenoic plasma concentration was associated with decreased odds of Group 1 status [quartile 4 aOR: 0.41 (95% CI: 0.17, 0.95)].Higher plasma concentrations of SFAs and MUFAs, primary de novo lipogenesis products, were associated with elevated FPG and HbA1c in a nationally representative study population in the United States. Additional studies are necessary to elucidate potential causal relationships between FAs (from endogenous production and dietary consumption) and diabetogenic indicators, as well as clinical implications for managing diabetes and prediabetes.

Keyword: lipogenesis

Anti-lipoapoptotic effects of Alisma orientalis extract on non-esterified fatty -induced HepG2 cells.

Liver steatosis was caused by lipid accumulation in the liver. Alisma orientale (AO) is recognized as a promising candidate with therapeutic efficacy for the treatment of nonalcoholic fatty liver disease (NAFLD). HepG2 hepatocyte cell line is commonly used for liver disease cell model.The HepG2 cells were cultured with the NEFAs mixture (oleic and acids, 2:1 ratio) for 24\xa0h to induce hepatic steatosis. Then different doses of Alisma orientale extract (AOE) was treated to HepG2 for 24\xa0h. Incubated cells were used for further experiments.The AOE showed inhibitory effects on lipid accumulation in the Oil Red O staining and Nile red staining tests with no cytotoxicity at a concentration of 300\xa0μg/mL. Fatty synthase (FASN) and acetyl-CoA carboxylase 1 (ACC1) mRNA and protein expression level were down-regulated after AOE treatment. Bcl-2 associated X protein (Bax) and c-Jun N-terminal kinase (JNK) mRNA expression level were decreased as well as p-JNK (activated form of JNK), Bax, cleaved caspase-9, caspase-3 protein expression level. Anti-apopototic B-cell lymphoma 2 (Bcl-2) protein level increased after AOE treatment. In addition, inflammatory protein expression including p-p65, p65, COX-2 and iNOS were inhibited by AOE treatment.The results suggest that AOE has anti-steatosis effects that involve , anti-lipoapoptosis, and anti-inflammation in the NEFA-induced NAFLD pathological cell model.

Keyword: lipogenesis

Obesity reduces the pro-angiogenic potential of adipose tissue stem cell-derived extracellular vesicles (EVs) by impairing miR-126 content: impact on clinical applications.

Soluble factors and cell-derived extracellular vesicles (EVs) are crucial tissue repair mediators in cell-based therapy. In the present study, we investigate the therapeutic impact of EVs released by adipose tissue-derived stem cells (ASCs) recovered from obese subjects\' visceral and subcutaneous tissues.ASCs were recovered from 10 obese (oASCs) and 6 non-obese (nASCs) participants and characterized. In selected experiments, nASCs and oASCs were cultured with (PA) or high glucose (HG), respectively. EVs from obese (oEVs) and non-obese (nEVs) subjects\' visceral and subcutaneous ASCs were collected after ultracentrifugation and analyzed for their cargo: microRNA-126 (miR-126), vascular endothelial growth factor (VEGF), and matrix metalloproteinase 2 (MMP-2), and for their biological effects on endothelial cells (ECs). Western blotting analysis and loss- and gain-of function experiments were performed.oEVs show impaired angiogenic potential compared with nEVs. This effect depends on EV cargo: reduced content of VEGF, MMP-2 and, more importantly, miR-126. We demonstrate, using gain- and loss-of-function experiments, that this reduced miR-126 content leads to Spred1 upregulation and the inhibition of the extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase pathway in ECs. We also show that PA treatment of nASCs translates into the release of EVs that recapitulate oEV cargo. Moreover, HG treatment of oASCs further reduces miR-126 EV content and EV-mediated in vitro angiogenesis. Finally, impaired pro-angiogenic potential is also detected in EVs released from obese subcutaneous adipose tissue-derived ASCs.These results indicate that obesity impacts on EV pro-angiogenic potential and may raise concerns about the use of adipose tissue-derived EVs in cell-based therapy in the obese setting.

Keyword: lipogenesis

Fatty Acids Have Different Adipogenic Differentiation Potentials in Stromal Vascular Cells Isolated from Abdominal Fat in Laying Hens.

This study was conducted to examine the effects of fatty acids (FA) with/without chicken serum (CS) on the expression of adipogenic transcripts and in chicken stromal vascular cells (SVC). In experiment 1, SVC were grown in DMEM containing 10% FBS (Control) and treated with 300\xa0µM oleic (OLA)\xa0+\xa0FBS, linoleic (LNA)\xa0+\xa0FBS, (PAM)\xa0+\xa0FBS, or stearic (STA)\xa0+\xa0FBS for 48\xa0h. In experiment 2, cells were grown in DMEM containing 5% CS and treated with 300\xa0µM OLA (CS\xa0+\xa0OLA), PAM (CS\xa0+\xa0PAM), STA (CS\xa0+\xa0STA) or 200\xa0µM LNA (CS\xa0+\xa0LNA) for 48\xa0h. was determined using Oil Red O staining and glycerol-3-phosphate dehydrogenase (GPDH) activity. The proportion of OLA, PAM, or STA was increased (P\xa0<\xa00.05) in SVC grown in either FBS or CS with OLA, PAM or STA. was induced in FBS\xa0+\xa0OLA, FBS\xa0+\xa0LNA, FBS\xa0+\xa0PAM, FBS\xa0+\xa0STA, CS\xa0+\xa0OLA, CS\xa0+\xa0LNA, CS\xa0+\xa0PAM, or CS\xa0+\xa0SAT compared to FBS. GPDH activity was significantly higher in FBS\xa0+\xa0OLA and FBS\xa0+\xa0LNA than one in FBS. Compared to FBS, the expression of FABP4 mRNA increased (P\xa0<\xa00.05) in FBS\xa0+\xa0OLA, FBS\xa0+\xa0LNA, or FBS\xa0+\xa0PAM, whereas that of C/EBPα, C/EBPβ, and ATGL increased (P\xa0<\xa00.05) in FBS\xa0+\xa0OLA or FBS\xa0+\xa0LNA cells. Expression of FABP4 and C/EBPβ mRNA was higher in CS, CS\xa0+\xa0OLA, CS\xa0+\xa0LNA, CS\xa0+\xa0PAM, or CS\xa0+\xa0SAT compared with (FBS, whereas the expression of ATGL and C/EBPα was higher in CS, CS\xa0+\xa0OLA, or CS\xa0+\xa0LNA than FBS cells. In conclusion, these results showed that FA have different potentials to induce , LNA is the most potent among the tested FA, and these potentials can be improved in the presence of CS.

Keyword: lipogenesis

Eicosapentaenoic Potentiates Brown Thermogenesis through FFAR4-dependent Up-regulation of miR-30b and miR-378.

Emerging evidence suggests that n-3 polyunsaturated fatty acids (PUFA) promote brown adipose tissue thermogenesis. However, the underlying mechanisms remain elusive. Here, we hypothesize that n-3 PUFA promotes brown by modulating miRNAs. To test this hypothesis, murine brown preadipocytes were induced to differentiate the fatty acids of , oleate, or eicosapentaenoic (EPA). The increases of brown-specific signature genes and oxygen consumption rate by EPA were concurrent with up-regulation of miR-30b and 378 but not by oleate or . Next, we hypothesize that free fatty receptor 4 (Ffar4), a functional receptor for n-3 PUFA, modulates miR-30b and 378. Treatment of Ffar4 agonist (GW9508) recapitulated the thermogenic activation of EPA by increasing oxygen consumption rate, brown-specific marker genes, and miR-30b and 378, which were abrogated in Ffar4-silenced cells. Intriguingly, addition of the miR-30b mimic was unable to restore EPA-induced Ucp1 expression in Ffar4-depleted cells, implicating that Ffar4 signaling activity is required for up-regulating the brown adipogenic program. Moreover, blockage of miR-30b or 378 by locked nucleic inhibitors significantly attenuated Ffar4 as well as brown-specific signature gene expression, suggesting the signaling interplay between Ffar4 and miR-30b/378. The association between miR-30b/378 and brown thermogenesis was also confirmed in fish oil-fed C57/BL6 mice. Interestingly, the Ffar4 agonism-mediated signaling axis of Ffar4-miR-30b/378-Ucp1 was linked with an elevation of cAMP in brown adipocytes, similar to cold-exposed or fish oil-fed brown fat. Taken together, our work identifies a novel function of Ffar4 in modulating brown partly through a mechanism involving cAMP activation and up-regulation of miR-30b and miR-378.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: lipogenesis

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic -related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated inflammation.

Keyword: lipogenesis

Decreased MiR-155 Level in the Peripheral Blood of Non-Alcoholic Fatty Liver Disease Patients may Serve as a Biomarker and may Influence LXR Activity.

Obesity is now a common risk factor for non-alcoholic fatty liver disease (NAFLD). Thus, it is important to explore its underlying mechanisms.Total RNA was extracted from peripheral whole blood samples from 50 NAFLD patients and 50 healthy controls. In addition, human liver specimens were obtained through liver biopsies from NAFLD patients and healthy controls. The level of miRNA was studied using real-time PCR. The expression of lipogenic genes was analyzed using western blot, and a dual luciferase reporter assay was conducted to identify the possible target gene. Adenovirus vectors were injected into the tail vein of the high fat diet (HFD)-fed mice to study the role of miR-155 on lipid accumulation in vivo.The level of miR-155 was markedly reduced in the livers and peripheral blood of NAFLD patients compared with healthy controls. Upregulation of miR-155 decreased intracellular lipid content and the SREBP1 and FAS protein levels, while inhibition of miR-155 enhanced the intracellular lipid content. The dual luciferase reporter assay showed that Liver X receptor (LXR)α was the target gene of miR-155, and silencing miR-155 reduced the expression of SREBP1 and FAS. An in vivo study showed that upregulation of miR-155 decreased the hepatic lipid accumulation mainly by suppressing the LXRα-dependent lipogenic signaling pathway.In summary, decreased expression of miR-155 in the peripheral blood may be utilized as a potential novel biomarker for NAFLD screening mainly by targeting LXRα.© 2016 The Author(s) Published by S. Karger AG, Basel.

Keyword: lipogenesis

Protective effects of gomisin N against hepatic steatosis through AMPK activation.

Gomisin N (GN) is a phytochemical derived from Schisandra chinensis. It has been reported to exert a protective effect against hepatic steatosis by attenuating endoplasmic reticulum (ER) stress. However, the detailed mechanism by which GN inhibits hepatic steatosis remains to be elucidated. In this study, we examined whether GN activates AMP-activated protein kinase (AMPK) and exerts therapeutic effects on liver X receptor (LXR)- or (PA)-induced triglyceride (TG) accumulation in HepG2 cells. Furthermore, in\xa0vivo protective effects of GN against hepatic steatosis were assessed in high-fat diet (HFD)-induced obese mice. GN stimulated the phosphorylation of AMPK, acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein 1c (SREBP1c) in HepG2 cells. It decreased the expression of genes, but increased the expression of fatty oxidation genes. Additionally, GN decreased the expression of genes induced by the LXR agonist T0901317 or PA in HepG2 cells, resulting in reduced intracellular TG content. However, preincubation with compound C, an AMPK inhibitor, prevented GN-mediated effects. Administration of GN to HFD-induced obese mice decreased HFD-induced liver weight, hepatic TG accumulation, and cytoplasmic lipid droplet. These findings demonstrate that GN activates the AMPK pathway and ameliorates HFD-induced hepatic steatosis.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Effects of Stanniocalcin-1 on glucose flux in rat brown adipose tissue.

The present work assesses in\xa0vitro the role of human Stanniocalcin 1 (hSTC-1) in C-glucose metabolism in brown adipose tissue (BAT) from fed rat. In the fed state, hSTC-1 decreases the incorporation of C from glucose into lipids in the rat BAT. The data support the hypothesis that the capacity of the glycerol-3-phosphate (G3P)-generating pathway (glycolysis) from glucose is regulated by hSTC-1, decreasing the adequate supply of G3P needed for fatty esterification and triacylglycerol (TG) storage in BAT. The results also suggest the effect of hSTC-1 on de novo fatty synthesis from pyruvate generated by C-glucose in the glycolysis pathway. In addition, by decreasing , hSTC-1 increased ATP levels and these two factors may decrease BAT thermogenic function. The presence of hSTC-1 in the incubation medium did not alter C-glucose and C-1- oxidation. The uncoupling protein 1 (UCP-1) expression was not altered by hSTC-1 either. In conclusion, hSTC-1 is one of the hormonal factors that control glucose metabolism in BAT in the fed state. The decrease of TG capacity synthesis from C-glucose by hSTC-1 compromises the BAT thermogenic capacity. Furthermore, the increase in ATP levels would inhibit a futile cycle via UCP-1, which dissipates oxidative energy as heat.Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: lipogenesis

Blood Fatty Profiles: New Biomarkers for Cardiometabolic Disease Risk.

Fatty (FA) profiles in different blood compartments are reflections of both diet and metabolism, and some FA levels are related to disease risk.Perhaps the most studied FA-disease relationship is between long-chain omega-3 polyunsaturated fatty acids [eicosapentaenoic (EPA) and docosahexaenoic (DHA)] and cardiovascular disease (CVD). Despite null results from recent large omega-3 FA supplementation trials, new research continues to support past studies showing that blood levels of EPA\u2009+\u2009DHA are inversely related to risk for total mortality and fatal CVD events. But blood levels of other FAs may also be useful markers of risk for a variety of diseases. The essential omega-6 FA linoleic is inversely associated with risk for developing type 2 diabetes (T2D), whereas risk for T2D is directly related to biomarkers of de novo ( and palmitoleic acids). Levels of industrially produced trans FAs have been linked to higher risk for CVD. Thus, blood levels of several individual FAs are emerging as modifiable biomarkers for risk of major chronic diseases.

Keyword: lipogenesis

Fatty 2-hydroxylase mediates diffusional mobility of Raft-associated lipids, GLUT4 level, and in 3T3-L1 adipocytes.

Straight chain fatty alpha-oxidation increases during differentiation of 3T3-L1 adipocytes, leading to a marked accumulation of odd chain length fatty acyl moieties. Potential roles of this pathway in adipocyte differentiation and are unknown. Mammalian fatty 2-hydroxylase (FA2H) was recently identified and suggested to catalyze the initial step of straight chain fatty alpha-oxidation. Accordingly, we examined whether FA2H modulates adipocyte differentiation and in mature adipocytes. FA2H level markedly increases during differentiation of 3T3-L1 adipocytes, and small interfering RNAs against FA2H inhibit the differentiation process. In mature adipocytes, depletion of FA2H inhibits basal and insulin-stimulated glucose uptake and , which are partially rescued by the enzymatic product of FA2H, 2-hydroxy . Expression of fatty- synthase and SCD1 was decreased in FA2H-depleted cells, and levels of GLUT4 and insulin receptor proteins were reduced. 2-Hydroxy fatty acids are enriched in cellular sphingolipids, which are components of membrane rafts. Accelerated diffusional mobility of raft-associated lipids was shown to enhance degradation of GLUT4 and insulin receptor in adipocytes. Consistent with this, depletion of FA2H appeared to increase raft lipid mobility as it significantly accelerated the rates of fluorescence recovery after photobleaching measurements of lipid rafts labeled with Alexa 488-conjugated cholera toxin subunit B. Moreover, the enhanced recovery rates were partially reversed by treatment with 2-hydroxy . In conclusion, our findings document the novel role of FA2H in adipocyte possibly by modulation of raft fluidity and level of GLUT4.

Keyword: lipogenesis

Determination of Fat Accumulation Reduction by Edible Fatty Acids and Natural Waxes .

Natural edible waxes mixed with plant oils, containing high levels of unsaturated fatty acids (FAs), are known as oleogels. Oleogels are used for replacing saturated FAs in animal-derived food with unsaturated FAs. However, the health effects of edible waxes are not yet clearly defined. The purpose of this study was to investigate the effect of FAs and natural waxes on the in 3T3-L1 cells. The 3T3-L1 cells were differentiated and treated with FAs and waxes. These FAs [ (PA), Stearic (SA), Oleic (OA), Linoleic (LA), and Alpha-linolenic (ALA)] and waxes [beeswax (BW) and carnauba wax (CW)] were prepared at varying concentrations, and cell toxicity, triglyceride accumulation, lipid droplets size, and distribution inside of cells were determined. Adipogenic gene expression including , , , , and was determined. Results showed that increasing the concentration of FAs and waxes led to a decrease in the adipocyte cells viability and metabolic performance. SA showed the highest level of triglyceride accumulation (p<0.05), whereas ALA showed the lowest (p<0.05). Both BW and CW at 3.0 ppm showed significantly higher lipid accumulation than in the control and other groups (p<0.05). ALA had significantly downregulated adipogenic gene expression levels, excluding those of , compared to the other treatment groups (p<0.05). Moreover, BW demonstrated similar adipogenic gene expression levels as ALA compared to CW. Consequently, ALA and BW may have health benefits by reducing and can be used in processed meat.

Keyword: lipogenesis

Effect of conjugated linoleic and acetate on milk fat synthesis and adipose in lactating dairy cows.

During biohydrogenation-induced milk fat depression (MFD), nutrients are spared from milk fat synthesis and are available for other metabolic uses. Acetate is the major carbon source spared and it may increase lipid synthesis in adipose tissue during MFD. The objective of this study was to compare the effect of trans-10,cis-12 conjugated linoleic (CLA) and the amount of acetate spared during CLA-induced MFD on adipose tissue . Nine multiparous, lactating, ruminally cannulated Holstein cows (244 ± 107 d in milk; 25 ± 8.4 kg of milk/d; mean ± standard deviation) were randomly assigned to treatments in a 3 × 3 Latin square design. Experimental periods were 4 d followed by a 10-d washout. Treatments were control (CON), ruminal infusion of acetate (AC; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), or abomasal infusion of CLA (10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA). Dry matter intake, milk yield, and milk protein yield and percentage were not affected by treatments. Compared with CON, milk fat yield decreased 23% and fat percent decreased 28% in CLA, and milk fat yield increased 20% in AC. Concentration and yield of milk de novo synthesized fatty acids (C16) was increased by CLA, compared with CON. Yield of de novo synthesized fatty acids and was increased by AC, compared with CON. capacity of adipose tissue explants was decreased 72% by CLA, but was not affected by AC. Acetate oxidation by adipose explants was not affected by treatments. Treatments had no effect on expression of key lipogenic factors, lipogenic enzymes, and leptin; however, expression of fatty binding protein 4 was reduced in CLA compared with CON. Additionally, hormone-sensitive lipase and perilipin 1 were decreased by CLA and acetate. Plasma glucose and glucagon concentrations were not affected by treatments; however, CLA increased nonesterified fatty acids 17.7%, β-hydroxybutyrate 16.1%, and insulin 27.8% compared with CON, and AC increased plasma β-hydroxybutyrate 18%. In conclusion, during CLA-induced MFD in low-producing cow adipose tissue was sensitive to the anti-lipogenic effects of CLA, while spared acetate did not stimulate adipose . However, acetate may play an important role in stimulating and improving energy status in the mammary gland under normal conditions.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: lipogenesis

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited -related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: lipogenesis

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: lipogenesis

α/β-hydrolase domain containing protein 15 (ABHD15)--an adipogenic protein protecting from apoptosis.

Our knowledge about adipocyte metabolism and development is steadily growing, yet many players are still undefined. Here, we show that α/β-hydrolase domain containing protein 15 (Abhd15) is a direct and functional target gene of peroxisome proliferator-activated receptor gamma (PPARγ), the master regulator of . In line, Abhd15 is mainly expressed in brown and white adipose tissue and strongly upregulated during in various murine and human cell lines. Stable knockdown of Abhd15 in 3T3-L1 cells evokes a striking differentiation defect, as evidenced by low lipid accumulation and decreased expression of adipocyte marker genes. In preconfluent cells, knockdown of Abhd15 leads to impaired proliferation, which is caused by apoptosis, as we see an increased SubG1 peak, caspase 3/7 activity, and BAX protein expression as well as a reduction in anti-apoptotic BCL-2 protein. Furthermore, apoptosis-inducing amounts of evoke a massive increase of Abhd15 expression, proposing an apoptosis-protecting role for ABHD15. On the other hand, in mature adipocytes physiological (i.e. non-apoptotic) concentrations of down-regulate Abhd15 expression. Accordingly, we found that the expression of Abhd15 in adipose tissue is reduced in physiological situations with high free fatty levels, like high-fat diet, fasting, and aging as well as in genetically obese mice. Collectively, our results position ABHD15 as an essential component in the development of adipocytes as well as in apoptosis, thereby connecting two substantial factors in the regulation of adipocyte number and size. Together with its intricate regulation by free fatty acids, ABHD15 might be an intriguing new target in obesity and diabetes research.

Keyword: lipogenesis

Altered composition of fatty acids exacerbates hepatotumorigenesis during activation of the phosphatidylinositol 3-kinase pathway.

Some clinical findings have suggested that systemic metabolic disorders accelerate in vivo tumor progression. Deregulation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway is implicated in both metabolic dysfunction and carcinogenesis in humans; however, it remains unknown whether the altered metabolic status caused by abnormal activation of the pathway is linked to the protumorigenic effect.We established hepatocyte-specific Pik3ca transgenic (Tg) mice harboring N1068fs*4 mutation.The Tg mice exhibited hepatic steatosis and tumor development. PPARγ-dependent was accelerated in the Tg liver, and the abnormal profile of accumulated fatty (FA) composition was observed in the tumors of Tg livers. In addition, the Akt/mTOR pathway was highly activated in the tumors, and in turn, the expression of tumor suppressor genes including Pten, Xpo4, and Dlc1 decreased. Interestingly, we found that the suppression of those genes and the enhanced in vitro colony formation were induced in the immortalized hepatocytes by the treatment with oleic (OA), which is one of the FAs that accumulated in tumors.Our data suggest that the unusual FA accumulation has a possible role in promoting in vivo hepato-tumorigenesis under constitutive activation of the PI3K pathway. The Pik3ca Tg mice might help to elucidate molecular mechanisms by which metabolic dysfunction contributes to in vivo tumor progression.Copyright © 2011 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Thioredoxin-interacting protein mediates hepatic and inflammation via PRMT1 and PGC-1α regulation in vitro and in vivo.

Non-alcoholic fatty liver disease (NAFLD) is strongly associated with obesity and type 2 diabetes. Thioredoxin-interacting protein (TXNIP) regulates the cellular redox state and metabolism and has been linked to many diseases, including diabetes. Therefore, we examined the role of TXNIP in hepatic steatosis in vitro and in vivo.Lipogenic and inflammatory proteins produced by hepatocytes treated with (PA) or transfected with TXNIP or Txnip siRNA were measured by Western blotting. Lipid accumulation was assessed using Oil Red O staining. Protein interactions were assessed by immunoprecipitation and proximity ligation assay. Hepatic protein levels were measured by Western blotting from wild type or Txnip(-/-) mice fed a high-fat diet (HFD) or chow diet. Livers from NAFLD patients were compared with normal liver by immunohistochemistry.PA increased TXNIP, and inflammatory and lipogenic proteins in both AML12 and H4IIE cells. It also increased the peroxisome proliferator-activated receptor gamma co-activator-1α (PGC-1α), which mediated the expression of lipogenic markers and lipid accumulation. In addition, PA increased protein arginine methyltransferase-1 (PRMT1) and PRMT1 siRNA abolished the increase in lipogenic markers with PGC-1α. Furthermore, TXNIP interacted with PRMT1 in PA-treated hepatocytes. In vivo, levels of lipogenic proteins, inflammatory molecules, PGC-1α, and PRMT1 were increased in the livers of HFD mice compared with those fed a chow diet, and were ameliorated in HFD Txnip(-/-) mice. Moreover, TXNIP, PRMT1, and PGC-1α were elevated in the livers of human NAFLD patients.TXNIP mediates hepatic via PRMT1 and PGC-1α regulation and inflammation in vitro and in vivo, implying that targeting TXNIP and PRMT1 is a potential therapeutic approach for treatment of NAFLD.Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: lipogenesis

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells.

4EBP1 is a chief downstream factor of mTORC1, and PPARγ is a key -related transcription factor. mTORC1 and PPARγ are associated with lipid metabolism. However, it is unknown which effector protein connects mTORC1 and PPARγ. This study investigated the interaction between 4EBP1 with PPARγ as part of the underlying mechanism by which insulin-induced lipid synthesis and secretion are regulated by mTORC1 in primary bovine mammary epithelial cells (pBMECs). Rapamycin, a specific inhibitor of mTORC1, downregulated 4EBP1 phosphorylation and the expression of PPARγ and the following lipogenic genes: lipin 1, DGAT1, ACC, and FAS. Rapamycin also decreased the levels of intracellular triacylglycerol (TAG); 10 types of fatty ; and the accumulation of TAG, (PA), and stearic (SA) in the cell culture medium. Inactivation of mTORC1 by shRaptor or shRheb attenuated the synthesis and secretion of TAG and PA. In contrast, activation of mTORC1 by Rheb overexpression promoted 4EBP1 phosphorylation and PPARγ expression and upregulated the mRNA and protein levels of lipin 1, DGAT1, ACC, and FAS, whereas the levels of intracellular and extracellular TAG, PA, and SA also rose. Further, 4EBP1 interacted directly with PPARγ. Inactivation of mTORC1 by shRaptor prevented the nuclear location of PPARγ. These results demonstrate that mTORC1 regulates lipid synthesis and secretion by inducing the expression of lipin 1, DGAT1, ACC, and FAS, which is likely mediated by the 4EBP1/PPARγ axis. This finding constitutes a novel mechanism by which lipid synthesis and secretion are regulated in pBMECs.

Keyword: lipogenesis

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo .

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, , and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver-body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: lipogenesis

Association between Nicotinamide Phosphoribosyltransferase and de novo in Nonalcoholic Fatty Liver Disease.

This study explored the association between serum nicotinamide phosphoribosyltransferase (NAMPT) and hepatic de novo (DNL) in nonalcoholic fatty liver disease (NAFLD) and determined whether or not this association is sex dependent.In this cross-sectional study, 62 consecutive patients (32 males, 30 females) with NAFLD were recruited. Serum NAMPT (by ELISA), , and the DNL index of erythrocyte membranes as markers of hepatic DNL (by gas chromatography) were analyzed. The controlled attenuation parameter (CAP) and body impedance analyzer were used to assess hepatic and body fat, respectively. Univariate and multiple linear regressions (to adjust for confounders) were used to analyze the association of serum NAMPT with , DNL index, CAP, and body fat.The respective values of serum NAMPT (2.44 ± 1.03 vs. 2.45 ± 1.13 ng/mL, p = 0.98), DNL index (3.11 [2.60-3.71] vs. 3.05 [2.40-3.59], p = 0.90), and (20.55% [15.34-24.04] vs. 22.64% [21.15-25.95], p = 0.07) were not significantly different between men and women, but those of CAP (326 [300-340] vs. 300 [261.25-329], p = 0.002) and body fat (37.71 ± 3.80 vs. 26.60 ± 5.70, p < 0.001) were significantly higher in women. In women, serum NAMPT had a significant negative association with the DNL index (β = -0.56, p = 0.01). The DNL index also had a significant negative association with body fat (β = -0.46, p = 0.02). In men, the only significant association was the positive association between serum NAMPT and CAP (β = 0.35, p = 0.035).Higher serum NAMPT in women was associated with a lower hepatic DNL index, while in men it was associated with higher hepatic fat and had no association with the DNL index. Therefore, the serum NAMPT level interpretation for NAFLD prognosis is probably sex dependent.© 2017 S. Karger AG, Basel.

Keyword: lipogenesis

miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 diabetes mellitus through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 diabetes mellitus (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid metabolism were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of lipid droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and lipid levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic lipid droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid metabolism dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid metabolism in T2DM, which can inhibit hepatic lipogenesis and and elevate glycogen synthesis by targeting STAT3.

Keyword: lipogenesis

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Metabolism Through Inhibition of the and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic metabolism. DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: lipogenesis

Regulation of peroxisome proliferator-activated receptor gamma on milk fat synthesis in dairy cow mammary epithelial cells.

Peroxisome proliferator-activated receptor gamma (PPARγ) participates in in rats, goats, and humans. However, the exact mechanism of PPARγ regulation on milk fat synthesis in dairy cow mammary epithelial cells (DCMECs) remains largely unexplored. The aim of this study was to investigate the role of PPARγ regarding milk fat synthesis in DCMECs and to ascertain whether milk fat precursor acetic and could interact with PPARγ signaling to regulate milk fat synthesis. For this study, we examined the effects of PPARγ overexpression and gene silencing on cell growth, triacylglycerol synthesis, and the messenger RNA (mRNA) and protein expression levels of genes involved in milk fat synthesis in DCMECs. In addition, we investigated the influences of acetic and on the mRNA and protein levels of milk lipogenic genes and triacylglycerol synthesis in DCMECs transfected with PPARγ small interfering RNA (siRNA) and PPARγ expression vector. The results showed that when PPARγ was silenced, cell viability, proliferation, and triacylglycerol secretion were obviously reduced. Gene silencing of PPARγ significantly downregulated the expression levels of milk fat synthesis-related genes in DCMECs. PPARγ overexpression improved cell viability, proliferation, and triacylglycerol secretion. The expression levels of milk lipogenic genes were significantly increased when PPARγ was overexpressed. Acetic and could markedly improve triacylglycerol synthesis and upregulate the expression levels of PPARγ and other lipogenic genes in DCMECs. These results suggest that PPARγ is a positive regulator of milk fat synthesis in DCMECs and that acetic and could partly regulate milk fat synthesis in DCMECs via PPARγ signaling.

Keyword: lipogenesis

GADD34-deficient mice develop obesity, nonalcoholic fatty liver disease, hepatic carcinoma and insulin resistance.

The prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing in parallel with the prevalence of obesity. DNA damage-inducible protein 34 (GADD34/Ppp1r15a), originally isolated from UV-inducible transcripts in Chinese hamster ovary (CHO) cells, dephosphorylates several kinases that function in important signaling cascades, including dephosphorylation of eIF2α. We examined the effects of GADD34 on natural life span by using GADD34-deficient mice. Here we observed for the first time that with age GADD34-deficient mice become obese, developing fatty liver followed by liver cirrhosis, hepatocellular carcinoma, and insulin resistance. We found that myofibroblasts and immune cells infiltrated the portal veins of aged GADD34-deficient mouse livers. A high-fat diet (HFD) induced a higher level of steatosis in young GADD34-deficient mice compared with WT mice. Differentiation into fat is dependent on insulin signaling. Insulin signaling in young GADD34-deficient mice was higher than that in WT mice, which explained the higher fat differentiation of mouse embryonic fibroblasts (MEFs) observed in GADD34-deficient mice. Through aging or a HFD, insulin signaling in GADD34-deficient liver converted to be down regulated compared with WT mice. We found that a HFD or palmitate treatment converted insulin signaling by up-regulating TNF-α and JNK.

Keyword: lipogenesis

Reduced miR-200b and miR-200c expression contributes to abnormal hepatic lipid accumulation by stimulating JUN expression and activating the transcription of srebp1.

Previous studies indicated that miR-200s participated in IL-6-induced hepatic insulin resistance. However, the role of miR-200s in hepatic lipid accumulation has not been elucidated. Here we found that miR-200b and miR-200c were reduced in the steatotic livers of mice fed a high-fat diet (HFD) and patients with nonalcoholic fatty liver disease. This down-regulation was accompanied by an increase in the expression of lipogenic proteins such as sterol regulatory element-binding protein 1 (SREBP1) and fatty synthase (FAS). The suppression of miR-200b and miR-200c in Hep1-6 and NCTC1469 hepatocytes enhanced intracellular triglyceride levels, which were associated with increased SREBP-1 and FAS protein levels. In contrast, the over-expression of miR-200b and miR-200c suppressed lipid accumulation and reduced the expression of SREBP1 and FAS in Hep1-6 and NCTC1469 cells transfected with miR-200b or miR-200c mimics. Importantly, the up-regulation of miR-200b and miR-200c could reverse oleic /-induced lipid accumulation in hepatocytes. A luciferase reporter assay identified that miR-200b and miR-200c could directly bind the 3\'UTR of jun. JUN activated the transcription of srebp1 to increase lipid accumulation. The data also demonstrated that increased miR-200b and miR-200c expression might be associated with sitagliptin-reduced hepatic lipid accumulation in mice fed a HFD. These findings suggest, for the first time, that reduced miR-200b and miR-200c expression contributes to abnormal hepatic lipid accumulation by stimulating JUN expression and activating the transcription of srebp1.

Keyword: lipogenesis

[Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: lipogenesis

Cis-9, trans-11 conjugated linoleic is endogenously synthesized from palmitelaidic (C16:1 trans-9) in bovine adipocytes.

Palmitelaidic (C16:1 trans-9) has been suggested to have beneficial effects on human health, including reduced adiposity. Objectives of this research were to quantify the amounts of palmitelaidic in beef samples and determine the effect of palmitelaidic supplementation on in bovine preadipocytes and adipocytes in vitro. For the first objective, palmitelaidic content of LM samples from steers finished on forage or concentrate systems was determined. Palmitelaidic in LM samples from forage-finished beef ranged from 10 to 17 mg/100 g of muscle corresponding to 0.52% to 0.65% of total fatty acids. Forage species grazed during finishing, and animal age at harvest also altered palmitelaidic concentrations and contents in the LM of forage-finished beef. Palmitelaidic concentration of concentrate-finished beef was lower (P < 0.05; 0.25% vs. 0.56%); however, because of increased (P < 0.05) total fatty content with concentrate finishing, amount of palmitelaidic was similar (P > 0.05) to beef from steers finished on pearl millet and greater (P < 0.05) than those finished on alfalfa. For the second objective, undifferentiated preadipocytes and differentiated adipocytes were supplemented with 0 to 300 μM of palmitelaidic . Palmitelaidic supplementation reduced (P < 0.05) cell viability of undifferentiated preadipocytes at greater levels (150 and 300 μM) but did not affect (P > 0.05) the viability of differentiated adipocytes. In preadipocytes, palmitelaidic increased (P < 0.05) palmitelaidic and trans-11 vaccenic (C18:1 trans-11) acids at high levels of supplementation (300 μM). In adipocytes, palmitelaidic supplementation increased (P < 0.05) palmitelaidic , trans-11 vaccenic , and total fatty content. In addition, cis-9, trans-11 CLA also increased (P < 0.05) with palmitelaidic supplementation in adipocytes. These results indicate that palmitelaidic can be elongated in both preadipocytes and adipocytes and desaturated in adipocytes to generate trans-11 vaccenic and cis-9, trans-11 CLA, respectively. Beef products are a source of palmitelaidic in the human diet, which can be elongated and desaturated to produce trans-11 vaccenic and cis-9, trans-11 CLA.

Keyword: lipogenesis

Prepartum fatty supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb liver and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb liver and adipose tissue fatty (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and liver and adipose tissue collected for FA and gene expression analysis. Liver concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in liver tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), fatty synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in liver of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain fatty 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that decreased, and lipolysis increased in lamb liver with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not liver, to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: lipogenesis

Fatty acids from fat cell lipolysis do not activate an inflammatory response but are stored as triacylglycerols in adipose tissue macrophages.

Activation of macrophages by fatty acids (FAs) is a potential mechanism linking obesity to adipose tissue (AT) inflammation and insulin resistance. Here, we investigated the effects of FAs released during adipocyte lipolysis on AT macrophages (ATMs).Human THP-1 macrophages were treated with media from human multipotent adipose-derived stem (hMADS) adipocytes stimulated with lipolytic drugs. Macrophages were also treated with mixtures of FAs and an inhibitor of Toll-like receptor 4, since this receptor is activated by saturated FAs. Levels of mRNA and the secretion of inflammation-related molecules were measured in macrophages. FA composition was determined in adipocytes, conditioned media and macrophages. The effect of chronic inhibition or acute activation of fat cell lipolysis on ATM response was investigated in vivo in mice.Whereas alone activates THP-1, conditioned media from hMADS adipocyte lipolysis had no effect on IL, chemokine and cytokine gene expression, and secretion by macrophages. Mixtures of FAs representing de novo or habitual dietary conditions also had no effect. FAs derived from adipocyte lipolysis were taken up by macrophages and stored as triacylglycerol droplets. In vivo, chronic treatment with an antilipolytic drug did not modify gene expression and number of ATMs in mice with intact or defective Tlr4. Stimulation of adipocyte lipolysis increased storage of neutral lipids by macrophages without change in number and phenotype.Our data suggest that adipocyte lipolysis does not activate inflammatory pathways in ATMs, which instead may act as scavengers of FAs.

Keyword: lipogenesis

Increased lipid synthesis and decreased β-oxidation in the liver of SHR/NDmcr-cp (cp/cp) rats, an animal model of metabolic syndrome.

SHR/NDmcr-cp (cp/cp) rats (SHR/NDcp) are an animal model of metabolic syndrome. A previous study of ours revealed drastic increases in the mass of (16:0), oleic (18:1n-9), palmitoleic (16:1n-7), cis-vaccenic (18:1n-7) and 5,8,11-eicosatrienoic acids in the liver of SHR/NDcp. However, detailed information on the class of lipid accumulated and the mechanism responsible for the overproduction of the accumulated lipid in the liver was not obtained. This study aimed to characterize the class of lipid accumulated and to explore the mechanism underlying the lipid accumulation in the liver of SHR/NDcp, in comparison with SHR/NDmcr-cp (+/+) (lean hypertensive littermates of SHR/NDcp) and Wistar Kyoto rats. In the liver of SHR/NDcp, de novo synthesis of fatty acids (16:0, 18:1n-9 and 16:1n-7) and triacylglycerol (TAG) synthesis were up-regulated and fatty β-oxidation was down-regulated. These perturbations of lipid metabolism caused fat accumulation in hepatocytes and accumulation of TAG, which were enriched with 16:0, 18:1n-9 and 16:1n-7, in the liver of SHR/NDcp. On the other hand, no changes were found in hepatic contents of diacylglycerol and unesterified fatty (FFA); among FFA, there were no differences in the hepatic concentrations of unesterified 16:0 and stearic between SHR/NDcp and two other groups of rats. Moreover, little change was brought about in the expression of genes responsive to endoplasmic reticulum stress in the liver of SHR/NDcp. These results may reinforce the pathophysiological role of stearoyl-CoA desaturase 1 and fatty elongase 6 in the liver of SHR/NDcp.

Keyword: lipogenesis

De novo in Atlantic salmon adipocytes.

Carnivorous teleost fish utilize glucose poorly, and the reason for this is not known. It is possible that the capacity of adipocytes to synthesize lipids from carbohydrate precursors through a process known as "de novo " (DNL) is one of the factors that contributes to glucose intolerance in Atlantic salmon.Primary adipocytes from Atlantic salmon differentiated in vitro were incubated with radiolabelled glucose in order to explore the capacity of salmon adipocytes to synthesize and deposit lipids from glucose through DNL. The lipid-storage capacity of adipocytes incubated with glucose was compared with that of cells incubated with the fatty . Quantitative PCR and immunohistochemistry were used to assess changes of genes and proteins involved in glucose and lipid transport and metabolism.Less than 0.1% of the radiolabelled glucose was metabolized to the fatty acids 16:0 and the stearoyl-CoA desaturase products 16:1 and 18:1 by DNL, whereas approximately 40% was converted to glycerol to form the triacylglycerol backbone of lipids. Transcriptional analysis indicated that adipocytes ensure the availability of necessary cofactors and other substrates for lipid synthesis and storage from glycolysis, the pentose phosphate pathway and glyceroneogenesis.We have shown for the first time that the DNL pathway is active in fish adipocytes. The capacity of the pathway to convert glucose into cellular lipids for storage is relatively low.The limited capacity of adipocytes to utilize glucose as a substrate for lipid deposition may contribute to glucose intolerance in salmonids.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Linking diet to acne metabolomics, inflammation, and comedogenesis: an update.

Acne vulgaris, an epidemic inflammatory skin disease of adolescence, is closely related to Western diet. Three major food classes that promote acne are: 1) hyperglycemic carbohydrates, 2) milk and dairy products, 3) saturated fats including trans-fats and deficient ω-3 polyunsaturated fatty acids (PUFAs). Diet-induced insulin/insulin-like growth factor (IGF-1)-signaling is superimposed on elevated IGF-1 levels during puberty, thereby unmasking the impact of aberrant nutrigenomics on sebaceous gland homeostasis. Western diet provides abundant branched-chain amino acids (BCAAs), glutamine, and . Insulin and IGF-1 suppress the activity of the metabolic transcription factor forkhead box O1 (FoxO1). Insulin, IGF-1, BCAAs, glutamine, and palmitate activate the nutrient-sensitive kinase mechanistic target of rapamycin complex 1 (mTORC1), the key regulator of anabolism and . FoxO1 is a negative coregulator of androgen receptor, peroxisome proliferator-activated receptor-γ (PPARγ), liver X receptor-α, and sterol response element binding protein-1c (SREBP-1c), crucial transcription factors of sebaceous . mTORC1 stimulates the expression of PPARγ and SREBP-1c, promoting sebum production. SREBP-1c upregulates stearoyl-CoA- and Δ6-desaturase, enhancing the proportion of monounsaturated fatty acids in sebum triglycerides. Diet-mediated aberrations in sebum quantity (hyperseborrhea) and composition (dysseborrhea) promote Propionibacterium acnes overgrowth and biofilm formation with overexpression of the virulence factor triglyceride lipase increasing follicular levels of free palmitate and oleate. Free palmitate functions as a "danger signal," stimulating toll-like receptor-2-mediated inflammasome activation with interleukin-1β release, Th17 differentiation, and interleukin-17-mediated keratinocyte proliferation. Oleate stimulates P. acnes adhesion, keratinocyte proliferation, and comedogenesis via interleukin-1α release. Thus, diet-induced metabolomic alterations promote the visible sebofollicular inflammasomopathy acne vulgaris. Nutrition therapy of acne has to increase FoxO1 and to attenuate mTORC1/SREBP-1c signaling. Patients should balance total calorie uptake and restrict refined carbohydrates, milk, dairy protein supplements, saturated fats, and trans-fats. A paleolithic-like diet enriched in vegetables and fish is recommended. Plant-derived mTORC1 inhibitors and ω-3-PUFAs are promising dietary supplements supporting nutrition therapy of acne vulgaris.

Keyword: lipogenesis

Nuclear factor-κB is a common upstream signal for growth differentiation factor-5 expression in brown adipocytes exposed to pro-inflammatory cytokines and palmitate.

We have previously demonstrated that genetic and acquired obesity similarly led to drastic upregulation in brown adipose tissue (BAT), rather than white adipose tissue, of expression of both mRNA and corresponding protein for the bone morphogenic protein/growth differentiation factor (GDF) member GDF5 capable of promoting brown . In this study, we evaluated expression profiles of GDF5 in cultured murine brown pre-adipocytes exposed to pro-inflammatory cytokines and free fatty acids (FFAs), which are all shown to play a role in the pathogenesis of obesity. Both interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were effective in up-regulating GDF5 expression in a concentration-dependent manner, while similar upregulation was seen in cells exposed to the saturated FFA palmitate, but not to the unsaturated FFA oleate. In silico analysis revealed existence of the putative nuclear factor-κB (NF-κB) binding site in the 5\'-flanking region of mouse GDF5, whereas introduction of NF-κB subunits drastically facilitated both promoter activity and expression of GDF5 in brown pre-adipocytes. Chromatin immunoprecipitation analysis confirmed significant facilitation of the recruitment of NF-κB to the GDF5 promoter in lysed extracts of BAT from leptin-deficient ob/ob obese mice. Upregulation o GDF5 expression was invariably inhibited by an NF-κB inhibitor in cultured brown pre-adipocytes exposed to IL-1β, TNF-α and palmitate. These results suggest that obesity leads to upregulation of GDF5 expression responsible for the promotion of brown through a mechanism relevant to activation of the NF-κB pathway in response to particular pro-inflammatory cytokines and/or saturated FFAs in BAT.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Palmitoleic (n-7) increases white adipocyte lipolysis and lipase content in a PPARα-dependent manner.

We investigated whether palmitoleic , a fatty that enhances whole body glucose disposal and suppresses hepatic steatosis, modulates triacylglycerol (TAG) metabolism in adipocytes. For this, both differentiated 3T3-L1 cells treated with either palmitoleic (16:1n7, 200 μM) or (16:0, 200 μM) for 24 h and primary adipocytes from wild-type or PPARα-deficient mice treated with 16:1n7 (300 mg·kg(-1)·day(-1)) or oleic (18:1n9, 300 mg·kg(-1)·day(-1)) by gavage for 10 days were evaluated for lipolysis, TAG, and glycerol 3-phosphate synthesis and gene and protein expression profile. Treatment of differentiated 3T3-L1 cells with 16:1n7, but not 16:0, increased basal and isoproterenol-stimulated lipolysis, mRNA levels of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) and protein content of ATGL and pSer(660)-HSL. Such increase in lipolysis induced by 16:1n7, which can be prevented by pharmacological inhibition of PPARα, was associated with higher rates of PPARα binding to DNA. In contrast to lipolysis, both 16:1n7 and 16:0 increased fatty incorporation into TAG and glycerol 3-phosphate synthesis from glucose without affecting glyceroneogenesis and glycerokinase expression. Corroborating in vitro findings, treatment of wild-type but not PPARα-deficient mice with 16:1n7 increased primary adipocyte basal and stimulated lipolysis and ATGL and HSL mRNA levels. In contrast to lipolysis, however, 16:1n7 treatment increased fatty incorporation into TAG and glycerol 3-phosphate synthesis from glucose in both wild-type and PPARα-deficient mice. In conclusion, palmitoleic increases adipocyte lipolysis and lipases by a mechanism that requires a functional PPARα.

Keyword: lipogenesis

Reduction in liver fat by dietary MUFA in type 2 diabetes is helped by enhanced hepatic fat oxidation.

The aim of this work was to investigate hepatic lipid metabolic processes possibly involved in the reduction of liver fat content (LF) observed in patients with type 2 diabetes after an isoenergetic diet enriched in monounsaturated fatty acids (MUFAs).This is an ancillary analysis of a published study. In a parallel-group design, 30 men and eight women, aged 35-70\xa0years, with type 2 diabetes and whose blood glucose was controlled satisfactorily (HbA\u2009<\u20097.5% [58\xa0mmol/mol]) by diet or diet plus metformin, were randomised by MINIM software to follow either a high-carbohydrate/high-fibre/low-glycaemic index diet (CHO/fibre diet, n\u2009=\u200920) or a high-MUFA diet (MUFA diet, n\u2009=\u200918) for 8\xa0weeks. The assigned diets were known for the participants and blinded for people doing measurements. Before and after intervention, LF was measured by H-MRS (primary outcome) and indirect indices of de novo (DNL) (serum triacylglycerol :linoleic ratio), stearoyl-CoA desaturase activity (SCD-1) (serum triacylglycerol palmitoleic: ratio) and hepatic β-oxidation of fatty acids (β-hydroxybutyrate plasma concentrations) were measured.LF was reduced by 30% after the MUFA diet, as already reported. Postprandial β-hydroxybutyrate incremental AUC (iAUC) was significantly less suppressed after the MUFA diet (n\u2009=\u200916) (-2504\u2009±\u20094488\xa0μmol/l\u2009×\u2009360\xa0min vs baseline -9021\u2009±\u20096489\xa0μmol/l\u2009×\u2009360\xa0min) while it was unchanged after the CHO/fibre diet (n\u2009=\u200917) (-8168\u2009±\u20099827\xa0μmol/l\u2009×\u2009360\xa0min vs baseline -7206\u2009±\u200910,005\xa0μmol/l\u2009×\u2009360\xa0min, p\u2009=\u20090.962) (mean\u2009±\u2009SD, p\u2009=\u20090.043). In the participants assigned to the MUFA diet, the change in postprandial β-hydroxybutyrate iAUC was inversely associated with the change in LF (r\u2009=\u2009-0.642, p\u2009=\u20090.010). DNL and SCD-1 indirect indices did not change significantly after either of the dietary interventions.Postprandial hepatic oxidation of fatty acids is a metabolic process possibly involved in the reduction of LF by a MUFA-rich diet in patients with type 2 diabetes.ClinicalTrials.gov FUNDING : The study was funded by Ministero Istruzione Università e Ricerca and Italian Minister of Health.

Keyword: lipogenesis

Markers of de novo in adipose tissue: associations with small adipocytes and insulin sensitivity in humans.

Previous studies have shown relationships between fatty ratios in adipose tissue triacylglycerol (TG), adipocyte size and measures of insulin sensitivity. We hypothesised that variations in adipose tissue de novo (DNL) in relation to adiposity might explain some of these observations.In a cross-sectional study, subcutaneous abdominal adipose tissue biopsies from 59 people were examined in relation to fasting and post-glucose insulin sensitivity. Adipocyte size, TG fatty composition and mRNA expression of lipogenic genes were determined.We found strong positive relationships between adipose tissue TG content of the fatty acids myristic (14:0) and stearic (18:0) with insulin sensitivity (HOMA model) (p < 0.01 for each), and inverse relationships with adipocyte size (p < 0.01, p < 0.05, respectively). Variation in 18:0 content was the determinant of the adipose tissue TG 18:1 n-9/18:0 ratio, which correlated negatively with insulin sensitivity (p < 0.01), as observed previously. Adipose tissue 18:0 content correlated positively with the mRNA expression of lipogenic genes (e.g. FASN, p < 0.01). Lipogenic gene expression (a composite measure derived from principal components analysis) was inversely correlated with adipocyte cell size (p < 0.001). There was no relationship between dietary saturated fatty intake and adipose tissue 18:0 content.Our data suggest a physiological mechanism whereby DNL is downregulated as adipocytes expand. Taken together with other data, they also suggest that hepatic and adipose tissue DNL are not regulated in parallel. We also confirm a strong relationship between small adipocytes and insulin sensitivity, which is independent of BMI.

Keyword: lipogenesis

Effects of berberine and cinnamic on -induced intracellular triglyceride accumulation in NIT-1 pancreatic β cells.

To investigate the effects of berberine (BBR) and cinnamic (CA), the main active components in Jiaotai Pill (, JTP), on (PA)-induced intracellular triglyceride (TG) accumulation in NIT-1 pancreatic β cells.Cells were incubated in culture medium containing PA (0.25 mmol/L) for 24 h. Then treatments with BBR (10 μmol/L), CA (100 μmol/L) and the combination of BBR and CA (BBR+CA) were performed respectively. Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by colorimetric assay. The expression of adenosine monophosphate-activated protein kinase (AMPK) protein and its downstream lipogenic and fatty oxidation genes, including fatty synthase (FAS), acetyl-coA carboxylase (ACC), phosphorylation acetyl-coA carboxylase (pACC), carnitine acyl transferase 1 (CPT-1) and sterol regulating element binding protein 1c (SREBP-1c) were determined by Western blot or real time polymerase chain reaction.PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 cells. PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as pACC and CPT-1. Meanwhile, AMPK downstream lipogenic genes including SREBP-1c mRNA, FAS and ACC protein expressions were increased. Treatments with BBR and BBR+CA, superior to CA, significantly reversed the above genes changes in NIT-1 pancreatic β cells. However, the synergistic effect of BBR and CA on intracellular TG content was not observed in the present study.It can be concluded that in vitro, BBR and BBR+CA could inhibit PA-induced lipid accumulation by decreasing and increasing lipid oxidation in NIT-1 pancreatic β cells.

Keyword: lipogenesis

Nrf2-Mediated Antioxidant Defense and Peroxiredoxin 6 Are Linked to Biosynthesis of Ester of 9-Hydroxystearic .

Fatty esters of hydroxy fatty acids (FAHFAs) are lipid mediators with promising antidiabetic and anti-inflammatory properties that are formed in white adipose tissue (WAT) via de novo , but their biosynthetic enzymes are unknown. Using a combination of lipidomics in WAT, quantitative trait locus mapping, and correlation analyses in rat BXH/HXB recombinant inbred strains, as well as response to oxidative stress in murine models, we elucidated the potential pathway of biosynthesis of several FAHFAs. Comprehensive analysis of WAT samples identified ∼160 regioisomers, documenting the complexity of this lipid class. The linkage analysis highlighted several members of the nuclear factor, erythroid 2 like 2 ()-mediated antioxidant defense system (), lipid-handling proteins (), and the family of flavin containing monooxygenases () as the positional candidate genes. Transgenic expression of and deletion of genes resulted in reduction of ester of 9-hydroxystearic (9-PAHSA) and 11-PAHSA levels, while oxidative stress induced by an inhibitor of glutathione synthesis increased PAHSA levels nonspecifically. Our results indicate that the synthesis of FAHFAs via carbohydrate-responsive element-binding protein-driven de novo depends on the adaptive antioxidant system and suggest that FAHFAs may link activity of this system with insulin sensitivity in peripheral tissues.© 2018 by the American Diabetes Association.

Keyword: lipogenesis

Increasing intake enhances milk production and prevents glucose-stimulated fatty disappearance without modifying systemic glucose tolerance in mid-lactation dairy cows.

Feeding saturated fatty acids may enhance milk yield in part by decreasing insulin sensitivity and shifting glucose utilization toward the mammary gland. Our objective was to evaluate the effects of (C16:0) on milk production and insulin sensitivity in cows. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows received a common sorghum silage-based diet and were randomly assigned to a diet containing no supplemental fat (control; n=10; 138±45d in milk) or C16:0 at 4% of ration DM (PALM; 98% C16:0; n=10; 136±44d in milk). Blood and milk were collected at routine intervals. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield by wk 7. Furthermore, PALM increased milk fat yield and energy-corrected milk at wk 3 and 7. Changes in milk production occurred in parallel with enhanced energy intake. Increased milk fat yield during PALM treatment was due to increased C16:0 and C16:1 incorporation; PALM had no effect on concentration of milk components, BW, or body condition score. Two weeks posttreatment, energy-corrected milk and milk fat yield remained elevated in PALM-fed cows whereas yields of milk were similar between treatments. Increased milk fat yield after PALM treatment was due to increased de novo and uptake of preformed fatty acids. The basal concentration of nonesterified fatty acids (NEFA) in plasma increased by d 4, 6, and 8 of PALM treatment, a response not observed thereafter. Although PALM supplementation did not modify insulin, glucose, or triacylglycerol levels in plasma, total cholesterol in plasma was elevated by wk 3. Estimated insulin sensitivity was lower during the first week of PALM treatment; however, glucose disposal following glucose tolerance tests was not modified. In contrast, C16:0 feeding reduced glucose-stimulated NEFA disappearance by wk 7. Results demonstrate that increasing dietary energy from C16:0 for 7wk improves milk yield and milk composition without modifying systemic glucose tolerance. Reduced glucose-stimulated NEFA disappearance with C16:0 feeding and elevated circulating NEFA may reflect changes in adipose tissue insulin sensitivity.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Protective effect of resveratrol derivatives on high-fat diet induced fatty liver by activating AMP-activated protein kinase.

Non-alcoholic fatty liver disease is associated with inhibited AMP-activated kinase (AMPK) and activation of sterol regulatory element binding protein 1 (SREBP-1). AMPK phosphorylation inhibits SREBP-1, a major transcription factor of de novo , by inhibiting the liver X receptor (LXR) or by direct phosphorylation. Resveratrol, a polyphenol, has regulatory effects on hepatic lipid metabolism as a potent AMPK activator. In this study, we evaluated the anti-steatogenic effects of resveratrol and its derivatives and identified the molecular mechanism in vitro and in vivo. Resveratrol and its derivatives decreased lipid accumulation by free fatty acids (FFA mixture; 0.5 mM, oleic : = 2: 1) in H4IIEC3 cells. Synthesized derivatives of resveratrol had lower cytotoxicity than the parental molecule with similar potency. SY-102 suppressed SREBP-1 maturation by T0901317, an LXR agonist, and decreased SRE luciferase activity and the mRNA levels of lipogenic genes. Inhibition of AMPK by pre-treatment with compound C completely blocked the effects of SY-102. To evaluate their efficacy in vivo, mice were fed a high-fat diet for 5 days, and resveratrol or SY-102 was administered orally for the last 2 days. Oral administration of the SY-102 increased AMPK phosphorylation, followed by reduced hepatic triglyceride accumulation to a similar extent as resveratrol. These data demonstrate that SY-102, a synthesized derivative of resveratrol, might provide a promising therapeutic effect against fatty liver disease.

Keyword: lipogenesis

mTORC2 Regulates Lipogenic Gene Expression through PPAR to Control Lipid Synthesis in Bovine Mammary Epithelial Cells.

The mechanistic target of rapamycin complex 2 (mTORC2) primarily functions as an effector of insulin/PI3K signaling to regulate cell proliferation and is associated with cell metabolism. However, the function of mTORC2 in lipid metabolism is not well understood. In the present study, mTORC2 was inactivated by the ATP-competitive mTOR inhibitor AZD8055 or shRNA targeting in primary bovine mammary epithelial cells (pBMECs). MTT assay was performed to examine the effect of AZD8055 on cell proliferation. ELISA assay and GC-MS analysis were used to determine the content of lipid. The mRNA and protein expression levels were investigated by RT/real-time PCR and western blot analysis, respectively. We found that cell proliferation, mTORC2 activation, and lipid secretion were inhibited by AZD8055. was knocked down and mTORC2 activation was specifically attenuated by the shRNA. Compared to control cells, the expression of the transcription factor gene and the lipogenic genes , , , and was downregulated in silencing cells. As a result, the content of intracellular triacylglycerol (TAG), (PA), docosahexaenoic (DHA), and other 16 types of fatty was decreased in the treated cells; the accumulation of TAG, PA, and DHA in cell culture medium was also reduced. Overall, mTORC2 plays a critical role in regulating lipogenic gene expression, lipid synthesis, and secretion in pBMECs, and this process probably is through PPAR. This finding provides a model by which is regulated in pBMECs.

Keyword: lipogenesis

follows a different metabolic pathway than oleic in human skeletal muscle cells; lower lipolysis rate despite an increased level of adipose triglyceride lipase.

Development of insulin resistance is positively associated with dietary saturated fatty acids and negatively associated with monounsaturated fatty acids. To clarify aspects of this difference we have compared the metabolism of oleic (OA, monounsaturated) and acids (PA, saturated) in human myotubes. Human myotubes were treated with 100μM OA or PA and the metabolism of [(14)C]-labeled fatty was studied. We observed that PA had a lower lipolysis rate than OA, despite a more than two-fold higher protein level of adipose triglyceride lipase after 24h incubation with PA. PA was less incorporated into triacylglycerol and more incorporated into phospholipids after 24h. Supporting this, incubation with compounds modifying lipolysis and reesterification pathways suggested a less influenced PA than OA metabolism. In addition, PA showed a lower accumulation than OA, though PA was oxidized to a relatively higher extent than OA. Gene set enrichment analysis revealed that 24h of PA treatment upregulated and fatty β-oxidation and downregulated oxidative phosphorylation compared to OA. The differences in lipid accumulation and lipolysis between OA and PA were eliminated in combination with eicosapentaenoic (polyunsaturated fatty ). In conclusion, this study reveals that the two most abundant fatty acids in our diet are partitioned toward different metabolic pathways in muscle cells, and this may be relevant to understand the link between dietary fat and skeletal muscle insulin resistance.Copyright © 2012 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Zinc alpha2 glycoprotein alleviates -induced intracellular lipid accumulation in hepatocytes.

Zinc alpha2 glycoprotein (ZAG) plays an important role in stimulating fat mobilization and lipolysis in adipose tissue, but its role in hepatic lipid metabolism remains unclear. (PA) was used to stimulate HepG2 cells with ZAG overexpression or ZAG knock down (shRNA). Overexpression of ZAG significantly inhibited , promoted lipolysis and fatty β-oxidation, and attenuated PA-induced intracellular fat accumulation. Moreover, ZAG overexpression dramatically stimulated adiponectin expression in HepG2 cells. In contrast, knockdown of ZAG notably inhibited fatty β-oxidation, increased and lipid accumulation. Collectively, these data suggest that ZAG has the potential to alleviate hepatosteatosis, making it a promising therapeutic target for fatty liver.Copyright © 2016. Published by Elsevier Ireland Ltd.

Keyword: lipogenesis

: Physiological Role, Metabolism and Nutritional Implications.

(PA) has been for long time negatively depicted for its putative detrimental health effects, shadowing its multiple crucial physiological activities. PA is the most common saturated fatty accounting for 20-30% of total fatty acids in the human body and can be provided in the diet or synthesized endogenously via lipogenesis (DNL). PA tissue content seems to be controlled around a well-defined concentration, and changes in its intake do not influence significantly its tissue concentration because the exogenous source is counterbalanced by PA endogenous biosynthesis. Particular physiopathological conditions and nutritional factors may strongly induce DNL, resulting in increased tissue content of PA and disrupted homeostatic control of its tissue concentration. The tight homeostatic control of PA tissue concentration is likely related to its fundamental physiological role to guarantee membrane physical properties but also to consent protein palmitoylation, palmitoylethanolamide (PEA) biosynthesis, and in the lung an efficient surfactant activity. In order to maintain membrane phospholipids (PL) balance may be crucial an optimal intake of PA in a certain ratio with unsaturated fatty acids, especially PUFAs of both n-6 and n-3 families. However, in presence of other factors such as positive energy balance, excessive intake of carbohydrates (in particular mono and disaccharides), and a sedentary lifestyle, the mechanisms to maintain a steady state of PA concentration may be disrupted leading to an over accumulation of tissue PA resulting in dyslipidemia, hyperglycemia, increased ectopic fat accumulation and increased inflammatory tone via toll-like receptor 4. It is therefore likely that the controversial data on the association of dietary PA with detrimental health effects, may be related to an excessive imbalance of dietary PA/PUFA ratio which, in certain physiopathological conditions, and in presence of an enhanced DNL, may further accelerate these deleterious effects.

Keyword: lipogenesis

Metabolomic elucidation of the effects of media and carbon sources on fatty production by Yarrowia lipolytica.

Lipid production by oleaginous Yarrowia lipolytica depends highly on culture environments, such as carbon sources, carbon/nitrogen (C/N) ratios, types of media, and cellular growth phases. In this study, the effects of media and carbon sources on lipid and metabolite production were investigated by profiling fatty acids and intracellular metabolites of Y. lipolytica grown in various media. The highest total fatty yield 114.04\u202f±\u202f6.23\u202fmg/g dry cell weight was achieved by Y. lipolytica grown in minimal medium with glycerol (SCG) in the exponential phase. The high lipid production by Y. lipolytica in SCG was presumed to be due to the higher C/N ratio in SCG than in the complex media. Moreover, glycerol promoted lipid production better than glucose in both complex and minimal media because glycerol can easily incorporate into the core of triglycerides. Metabolite profiling revealed that levels of long-chain fatty acids, such as stearic , , and arachidic , increased in SCG medium. Meanwhile, in complex media supplemented with either glucose or glycerol, levels of amino acids, such as cysteine, methionine, and glycine, highly increased. This metabolomic approach could be applied to modulate the global metabolic network of Y. lipolytica for producing lipids and other valuable products.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

TRIF-dependent Toll-like receptor signaling suppresses transcription in hepatocytes and prevents diet-induced hepatic steatosis.

Nonalcoholic fatty liver disease (NAFLD) includes a spectrum of diseases that ranges in severity from hepatic steatosis to steatohepatitis, the latter of which is a major predisposing factor for liver cirrhosis and cancer. Toll-like receptor (TLR) signaling, which is critical for innate immunity, is generally believed to aggravate disease progression by inducing inflammation. Unexpectedly, we found that deficiency in TIR domain-containing adaptor-inducing interferon-β (TRIF), a cytosolic adaptor that transduces some TLR signals, worsened hepatic steatosis induced by a high-fat diet (HFD) and that such exacerbation was independent of myeloid cells. The aggravated steatosis in mice was due to the increased hepatocyte transcription of the gene encoding stearoyl-coenzyme A (CoA) desaturase 1 (SCD1), the rate-limiting enzyme for . Activation of the TRIF pathway by polyinosinic:polycytidylic [poly(I:C)] suppressed the increase in SCD1 abundance induced by or an HFD and subsequently prevented lipid accumulation in hepatocytes. Interferon regulatory factor 3 (IRF3), a transcriptional regulator downstream of TRIF, acted as a transcriptional suppressor by directly binding to the promoter. These results suggest an unconventional metabolic function for TLR/TRIF signaling that should be taken into consideration when seeking to pharmacologically inhibit this pathway.Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Keyword: lipogenesis

Differential alterations of the concentrations of endocannabinoids and related lipids in the subcutaneous adipose tissue of obese diabetic patients.

The endocannabinoids, anandamide and 2-AG, are produced by adipocytes, where they stimulate via cannabinoid CB1 receptors and are under the negative control of leptin and insulin. Endocannabinoid levels are elevated in the blood of obese individuals and nonobese type 2 diabetes patients. To date, no study has evaluated endocannabinoid levels in subcutaneous adipose tissue (SAT) of subjects with both obesity and type 2 diabetes (OBT2D), characterised by similar adiposity and whole body insulin resistance and lower plasma leptin levels as compared to non-diabetic obese subjects (OB).The levels of anandamide and 2-AG, and of the anandamide-related PPARalpha ligands, oleoylethanolamide (OEA) and palmitoylethanolamide (PEA), in the SAT obtained by abdominal needle biopsy in 10 OBT2D, 11 OB, and 8 non-diabetic normal-weight (NW) subjects, were measured by liquid chromatography-mass spectrometry. All subjects underwent a hyperinsulinaemic euglycaemic clamp.As compared to NW, anandamide, OEA and PEA levels in the SAT were 2-4.4-fold elevated (p < 0.05), and 2-AG levels 2.3-fold reduced (p < .05), in OBT2D but not in OB subjects. Anandamide, OEA and PEA correlated positively (p < .05) with SAT leptin mRNA and free fatty during hyperinsulinaemic clamp, and negatively with SAT LPL activity and plasma HDL-cholesterol, which were all specifically altered in OBT2D subjects.The observed alterations emphasize, for the first time in humans, the potential different role and regulation of adipose tissue anandamide (and its congeners) and 2-AG in obesity and type 2 diabetes.

Keyword: lipogenesis

Cancer cells incorporate and remodel exogenous palmitate into structural and oncogenic signaling lipids.

De novo is considered the primary source of fatty acids for lipid synthesis in cancer cells, even in the presence of exogenous fatty acids. Here, we have used an isotopic fatty labeling strategy coupled with metabolomic profiling platforms to comprehensively map incorporation into complex lipids in cancer cells. We show that cancer cells and tumors robustly incorporate and remodel exogenous palmitate into structural and oncogenic glycerophospholipids, sphingolipids, and ether lipids. We also find that fatty incorporation into oxidative pathways is reduced in aggressive human cancer cells, and instead shunted into pathways for generating structural and signaling lipids. Our results demonstrate that cancer cells do not solely rely on de novo , but also utilize exogenous fatty acids for generating lipids required for proliferation and protumorigenic lipid signaling. This article is part of a special issue entitled Lipid Metabolism in Cancer.© 2013.

Keyword: lipogenesis

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: lipogenesis

Quercetin inhibits fatty and triacylglycerol synthesis in rat-liver cells.

Quercetin plays a cardiovascular protective role because of its antioxidant capacity and ability to modulate dyslipidemia. As alterations in hepatic lipid synthesis are crucial to the regulation of serum lipid levels, we investigated the quercetin effect on in rat liver cells.The effect of quercetin on the rate of synthesis of fatty acids, cholesterol, neutral lipids, phospholipids and very-low-density lipoproteins (VLDL) was investigated in rat hepatocyte suspensions following [1-(14)C]acetate incorporation into these lipid fractions. Enzyme activities of acetyl-CoA carboxylase (ACC) and fatty synthase (FAS) as well as diacylglycerol acyltransferase (DGAT) and 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA-R), pace-setting steps of de novo fatty , triacylglycerol (TAG) and cholesterol synthesis respectively were assayed in digitonin-permeabilized hepatocytes.Within 30 min of quercetin addition to the hepatocytes, inhibition (IC50 approximately 25 microM) of fatty synthesis occurred. A reduction in label incorporation mainly into TAG was observed. Among neosynthesized fatty acids, formation was greatly reduced, suggesting that enzymatic step(s) of de novo fatty synthesis was affected. Only ACC activity was noticeably reduced, but no change in FAS activity was observed. DGAT activity was also inhibited. The decreased intracellular TAG content was paralleled by a reduction in acetate incorporation into VLDL-TAG. Conversely, cholesterol synthesis and HMG-CoA-R were not significantly affected by quercetin.In hepatocytes from normal rats, the quercetin-induced decrease in both de novo fatty and TAG synthesis, with a consequent reduction in VLDL-TAG formation, may represent a potential mechanism contributing to the reported hypotriacylglycerolemic effect of quercetin.

Keyword: lipogenesis

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic insulin resistance and Type 2 diabetes (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of insulin resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of lipid species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with insulin resistance and risk of developing T2D.

Keyword: lipogenesis

Genome-wide association study identifies novel loci associated with concentrations of four plasma phospholipid fatty acids in the de novo pathway: results from the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium.

BACKGROUND- (16:0), stearic (18:0), palmitoleic (16:1n-7), and oleic (18:1n-9) are major saturated and monounsaturated fatty acids that affect cellular signaling and metabolic pathways. They are synthesized via de novo and are the main saturated and monounsaturated fatty acids in the diet. Levels of these fatty acids have been linked to diseases including type 2 diabetes mellitus and coronary heart disease. METHODS AND RESULTS- Genome-wide association studies were conducted in 5 population-based cohorts comprising 8961 participants of European ancestry to investigate the association of common genetic variation with plasma levels of these 4 fatty acids. We identified polymorphisms in 7 novel loci associated with circulating levels of ≥1 of these fatty acids. ALG14 (asparagine-linked glycosylation 14 homolog) polymorphisms were associated with higher 16:0 (P=2.7×10(-11)) and lower 18:0 (P=2.2×10(-18)). FADS1 and FADS2 (desaturases) polymorphisms were associated with higher 16:1n-7 (P=6.6×10(-13)) and 18:1n-9 (P=2.2×10(-32)) and lower 18:0 (P=1.3×10(-20)). LPGAT1 (lysophosphatidylglycerol acyltransferase) polymorphisms were associated with lower 18:0 (P=2.8×10(-9)). GCKR (glucokinase regulator; P=9.8×10(-10)) and HIF1AN (factor inhibiting hypoxia-inducible factor-1; P=5.7×10(-9)) polymorphisms were associated with higher 16:1n-7, whereas PKD2L1 (polycystic kidney disease 2-like 1; P=5.7×10(-15)) and a locus on chromosome 2 (not near known genes) were associated with lower 16:1n-7 (P=4.1×10(-8)). CONCLUSIONS- Our findings provide novel evidence that common variations in genes with diverse functions, including protein-glycosylation, polyunsaturated fatty metabolism, phospholipid modeling, and glucose- and oxygen-sensing pathways, are associated with circulating levels of 4 fatty acids in the de novo pathway. These results expand our knowledge of genetic factors relevant to de novo and fatty biology.

Keyword: lipogenesis

[THE EXCESS OF FATTY IN FOOD AS MAIN CAUSE OF LIPOIDOSIS OF INSULIN-DEPENDENT CELLS: SKELETAL MYOCYTES, CARDIO-MYOCYTES, PERIPORTAL HEPATOCYTES, KUPFFER MACROPHAGES AND B-CELLS OF PANCREAS].

In phylogenesis, becoming of biologicalfunctions and biological reactions proceeds with the purpose ofpermanent increasing of "kinetic perfection ". The main role belongs to factors ofphysical, chemical and biological kinetics, their evaluation using systemic approach technique under permanent effect of natural selection. The late-in-phylogenesis insulin, proceeded with, in development of biological function of locomotion, specialization of insulin-dependent cells: skeletal myocytes, syncytium of cardiomyocytes, subcutaneous adipocytes, periportal hepatocytes, Kupffer\'s macrophages and β-cells of islets of pancreas. The insulin initiated formation of new, late in phylogenesis, large pool of fatty cells-subcutaneous adipocytes that increased kinetic parameters of biological function of locomotion. In realization of biological function of locomotion only adipocytes absorb exogenous mono unsaturated and saturated fatty acids in the form of triglycerides in composition of oleic and lipoproteins of very low density using apoE/B-100 endocytosis. The rest of insulin-dependent cells absorb fatty acids in the form of unesterified fatty acids from associates with albumin and under effect of CD36 of translocase offatty acids. The insulin in all insulin-depended cells inhibits biological reaction of lipolysis enhancing contributing into development of lipoidosis. The insulin expresses transfer offatty acids in the form of unsaturated fatty acids from adipocytes into matrix of mitochondria. The insulin supplies insulin-dependent cells with substrates for acquiring energy subject to that in pool of unsaturated fatty acids in adipocytes prevails hydrophobic unsaturated fatiy that slowly passes into matrix through external membrane ofmitochondria; oxidases of mitochondria so slowly implement its β-oxidation that content of exogenous unsaturatedfatty can\'t be higher than phylogenetic, physiological level - 15% of all amount offatty acids transferring to insulin-dependent cells. The insulin can\'t both to decrease content of exogenous fatty and inhibit lipolysis in visceral fatty cells of omentum.

Keyword: lipogenesis

Palmitoleic is elevated in fatty liver disease and reflects hepatic .

Biochemical evidence has linked the coordinate control of fatty (FA) synthesis with the activity of stearoyl-CoA desaturase-1 (SCD1). The ratio of 16:1n-7 to 16:0 [SCD1₁₆] in plasma triacylglycerol FA has been used as an index to reflect liver SCD1₁₆ activity and has been proposed as a biomarker of FA synthesis, although this use has not been validated by comparison with isotopically measured de novo (DNL(Meas)).We investigated plasma lipid 16:1n-7 and FA indexes of elongation and desaturation in relation to .In this cross-sectional investigation of metabolism, 24 overweight adults, who were likely to have elevated DNL, consumed D2O for 10 d and had liver fat (LF) measured by magnetic resonance spectroscopy. Very-low-density lipoprotein (VLDL)-triacylglycerols and plasma free FA [nonesterified fatty acids (NEFAs)] were analyzed by using gas chromatography for the FA composition (molar percentage) and gas chromatography-mass spectrometry and gas chromatography-combustion isotope ratio mass spectrometry for deuterium enrichment.In all subjects, VLDL-triacylglycerol 16:1n-7 was significantly (P < 0.01) related to DNL(Meas) (r = 0.56), liver fat (r = 0.53), and adipose insulin resistance (r = 0.56); similar positive relations were shown with the SCD1₁₆ index, and the pattern in NEFAs echoed that of VLDL-triacylglycerols. Compared with subjects with low LF (3.1 ± 2.7%; n = 11), subjects with high LF (18.4 ± 3.6%; n = 13) exhibited a 45% higher VLDL-triacylglycerol 16:1n-7 molar percentage (P < 0.01), 16% of subjects had lower 18:2n-6 (P = 0.01), and 27% of subjects had higher DNL as assessed by using a published DNL index (ratio of 16:0 to 18:2n-6; P = 0.03), which was isotopically confirmed by DNL(Meas) (increased 2.5-fold; P < 0.01). Compared with 16:0 in the diet, the low amount of dietary 16:1n-7 in VLDL-triacylglycerols corresponded to a stronger signal of elevated DNL.The current data provide support for the use of the VLDL-triacylglycerol 16:1n-7 molar percentage as a biomarker for elevated liver fat when isotope use is not feasible; however, larger-scale confirmatory studies are needed.ClinicalTrials.gov .© 2015 American Society for Nutrition.

Keyword: lipogenesis

Free fatty -induced histone acetyltransferase activity accelerates lipid accumulation in HepG2 cells.

Non-alcoholic fatty liver disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and lipid metabolism in the liver have not been thoroughly investigated.Following oleic- (OPA)-induced lipid accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between lipid accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and lipid accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: lipogenesis

Fatty acids in the de novo pathway and risk of coronary heart disease: the Cardiovascular Health Study.

De novo (DNL) is an endogenous pathway whereby carbohydrates and proteins are converted to fatty acids. DNL could affect coronary heart disease (CHD) or sudden cardiac arrest (SCA) via generation of specific fatty acids. Whether these fatty acids are prospectively associated with SCA or other CHD events is unknown.The objective was to investigate the relations of 4 fatty acids in the DNL pathway- (16:0), palmitoleic (16:1n-7), 7-hexadecenoic (16:1n-9), and cis-vaccenic (18:1n-7)-with incident CHD, including fatal CHD, nonfatal myocardial infarction (NFMI), and SCA.A community-based prospective study was conducted in 2890 men and women aged ≥65 y, who were free of known CHD at baseline and who were followed from 1992 to 2006. Cardiovascular disease risk factors and plasma phospholipid fatty acids were measured at baseline by using standardized methods. Incident CHD was ascertained prospectively and was centrally adjudicated by using medical records. Risk was assessed by using multivariable-adjusted Cox proportional hazards.During 29,835 person-years of follow-up, 631 CHD and 71 SCA events occurred. Both 18:1n-7 and 16:1n-9 were associated with a higher risk of SCA [multivariable-adjusted hazard ratio (95% CI) for the interquintile range: 7.63 (2.58, 22.6) for 18:1n-7 and 2.30 (1.16, 4.55) for 16:1n-9] but not of total CHD, fatal CHD, or NFMI. In secondary analyses censored to mid-follow-up (7 y) to minimize the effects of changes in concentrations over time, 16:1n-9 was also associated with a significantly higher risk of total CHD (2.11; 1.76, 2.54), including a higher risk of CHD death, NFMI, and SCA; 16:0 and 16:1n-7 were not associated with clinical CHD outcomes.Higher plasma phospholipid 18:1n-7 and 16:1n-9 concentrations were prospectively associated with an elevated risk of SCA but not of other CHD events, except in secondary analyses.

Keyword: lipogenesis

Palmitoleic (16:1n7) increases oxygen consumption, fatty oxidation and ATP content in white adipocytes.

We have recently demonstrated that palmitoleic (16:1n7) increases lipolysis, glucose uptake and glucose utilization for energy production in white adipose cells. In the present study, we tested the hypothesis that palmitoleic modulates bioenergetic activity in white adipocytes.For this, 3\xa0T3-L1 pre-adipocytes were differentiated into mature adipocytes in the presence (or absence) of (16:0) or palmitoleic (16:1n7) at 100 or 200\xa0μM. The following parameters were evaluated: lipolysis, , fatty (FA) oxidation, ATP content, oxygen consumption, mitochondrial mass, citrate synthase activity and protein content of mitochondrial oxidative phosphorylation (OXPHOS) complexes.Treatment with 16:1n7 during 9\xa0days raised basal and isoproterenol-stimulated lipolysis, FA incorporation into triacylglycerol (TAG), FA oxidation, oxygen consumption, protein expression of subunits representing OXPHOS complex II, III, and V and intracellular ATP content. These effects were not observed in adipocytes treated with 16:0.Palmitoleic , by concerted action on lipolysis, FA esterification, mitochondrial FA oxidation, oxygen consumption and ATP content, does enhance white adipocyte energy expenditure and may act as local hormone.

Keyword: lipogenesis

Increased FNDC5/Irisin expression in human hepatocellular carcinoma.

The fibronectin type III domain containing 5 (FNDC5)/Irisin, a novel energy-regulating hormone, is associated with lipid and carbohydrate metabolism. It is produced in low amounts by normal hepatic tissue, while in human hepatocellular carcinoma (HCC), in which aberrant de novo (DNL) occurs, the hepatic expression of FNDC5/Irisin is still unknown. The gene expression of FNDC5/Irisin, associated to key regulators of DNL, inflammation and cancer progression was evaluated in liver tissue of 18 patients with HCC undergoing liver transplantation and of 18 deceased donors. Hepatic mRNA expression of FNDC5/Irisin and stearoyl-CoA desaturase (SCD-1), main enzymatic regulator of DNL, were significantly higher in HCC patients than in donors (p<0.0001 and p=0.015, respectively). The hepatic mRNA expression of the neurogenic locus notch homolog protein 1 (NOTCH1) tended to be higher in HCC patients than in donors (p=0.06). Only in HCC patients, hepatic FNDC5/Irisin strongly correlated with the transcription factor sterol regulatory element-binding factor 1, SCD-1, NOTCH1, tumor necrosis factor-α and Interleukin-6 mRNA expression. Further, in HCC patients, FNDC5/Irisin mRNA tended to correlate to plasma lipid profile namely triglycerides, /linoleic and polyunsaturated fatty /saturated fatty ratios. In conclusion, HCC-liver tissue over-expressed FNDC5/Irisin in association with gene expression of mediators involved in , inflammation and cancer, suggesting a possible protective role of the hormone from the liver damage.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Global metabolic profiling of infection by an oncogenic virus: KSHV induces and requires for survival of latent infection.

Like cancer cells, virally infected cells have dramatically altered metabolic requirements. We analyzed global metabolic changes induced by latent infection with an oncogenic virus, Kaposi\'s Sarcoma-associated herpesvirus (KSHV). KSHV is the etiologic agent of Kaposi\'s Sarcoma (KS), the most common tumor of AIDS patients. Approximately one-third of the nearly 200 measured metabolites were altered following latent infection of endothelial cells by KSHV, including many metabolites of anabolic pathways common to most cancer cells. KSHV induced pathways that are commonly altered in cancer cells including glycolysis, the pentose phosphate pathway, amino production and fatty synthesis. Interestingly, over half of the detectable long chain fatty acids detected in our screen were significantly increased by latent KSHV infection. KSHV infection leads to the elevation of metabolites involved in the synthesis of fatty acids, not degradation from phospholipids, and leads to increased lipid droplet organelle formation in the infected cells. Fatty synthesis is required for the survival of latently infected endothelial cells, as inhibition of key enzymes in this pathway led to apoptosis of infected cells. Addition of to latently infected cells treated with a fatty synthesis inhibitor protected the cells from death indicating that the products of this pathway are essential. Our metabolomic analysis of KSHV-infected cells provides insight as to how oncogenic viruses can induce metabolic alterations common to cancer cells. Furthermore, this analysis raises the possibility that metabolic pathways may provide novel therapeutic targets for the inhibition of latent KSHV infection and ultimately KS tumors.

Keyword: lipogenesis

Lipid accumulation stimulates the cap-independent translation of SREBP-1a mRNA by promoting hnRNP A1 binding to its 5\'-UTR in a cellular model of hepatic steatosis.

Non-alcoholic fatty liver disease (NAFLD) is a chronic disease characterized by accumulation of lipid droplets in hepatocytes. Enhanced release of non-esterified fatty acids from adipose tissue accounts for a remarkable fraction of accumulated lipids. However, the de novo (DNL) is also implicated in the etiology of the NAFLD. Sterol Regulatory Element-Binding Protein-1 (SREBP-1) is a transcription factor modulating the expression of several lipogenic enzymes. In the present study, in order to investigate the effect of lipid droplet accumulation on DNL, we used a cellular model of steatosis represented by HepG2 cells cultured in a medium supplemented with free oleic and fatty acids (FFAs). We report that FFA supplementation induces the expression of genes coding for enzymes involved in the DNL as well as for the transcription factor SREBP-1a. The SREBP-1a mRNA translation, dependent on an internal ribosome entry site (IRES), and the SREBP-1a proteolytic cleavage are activated by FFAs. Furthermore, FFA treatment enhances the expression and the nucleus-cytosolic shuttling of hnRNP A1, a trans-activating factor of SREBP-1a IRES. The binding of hnRNP A1 to the SREBP-1a IRES is also increased upon FFA supplementation. The relocation of hnRNP A1 and the consequent increase of SREBP-1a translation are dependent on the p38 MAPK signal pathway, which is activated by FFAs. By RNA interference approach, we demonstrate that hnRNP A1 is implicated in the FFA-induced expression of SREBP-1a and of its target genes as well as in the lipid accumulation in cells.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Upregulation of lipid synthesis in small rat adipocytes by microvesicle-associated CD73 from large adipocytes.

Filling-up lipid stores is critical for size increase of mammalian adipocytes. The glycosylphosphatidylinositol (GPI)-anchored protein, CD73, is released from adipocytes into microvesicles in response to the lipogenic stimuli, palmitate, the antidiabetic sulfonylurea drug glimepiride, phosphoinositolglycans (PIG), and H(2)O(2). Upon incubation of microvesicles with adipocytes, CD73 is translocated to cytoplasmic lipid droplets (LD) and esterification is upregulated. The role of CD73-harboring microvesicles in coordinating esterification between differently sized adipocytes was studied here. Populations consisting of either small or large or of both small and large isolated rat adipocytes as well as native adipose tissue pieces from young and old rats were incubated with or depleted of endogenous microvesicles and analyzed for translocation of CD73 and esterification in response to the lipogenic stimuli. Large adipocytes exhibited higher and lower efficacy in releasing CD73 into microvesicles and in translocating CD73 to LD, respectively, compared to small adipocytes. Populations consisting of both small and large adipocytes were more active in esterification in response to the lipogenic stimuli than either small or large adipocytes. With both adipocytes and adipose tissue pieces from young rats esterification stimulation by the lipogenic stimuli was abrogated by depletion of CD73-harboring microvesicles from the incubation medium and interstitial spaces, respectively. In conclusion, stimulus-induced lipid synthesis between differently sized adipocytes is controlled by the release of microvesicle-associated CD73 from large cells and its subsequent translocation to LD of small cells. This information transfer via microvesicles harboring GPI-anchored proteins may shift the burden of triacylglycerol storage from large to small adipocytes.

Keyword: lipogenesis

Liver MicroRNA-291b-3p Promotes Hepatic through Negative Regulation of Adenosine 5\'-Monophosphate (AMP)-activated Protein Kinase α1.

In a microarray study, we found that hepatic miR-291b-3p was significantly increased in leptin-receptor-deficient type 2 mice (db/db), a mouse model of diabetes. The function of miR-291b-3p is unknown. The potential role of miR-291b-3p in regulating hepatic lipid metabolism was explored in this study. High-fat diet (HFD)- and chow-fed mice were injected with an adenovirus expressing a miR-291b-3p inhibitor and a miR-291b-3p mimic through the tail vein. Hepatic lipids and lipogenic gene expression were analyzed. Additionally, gain- and loss-of-function studies were performed in vitro to identify direct targets of miR-291b-3p. MiR-291b-3p expression and the protein levels of sterol regulatory element-binding protein 1 (SREBP1) and fatty synthase (FAS) were increased in the steatotic liver of db/db mice and HFD-fed mice versus their respective controls. Inhibition of hepatic miR-291b-3p expression prevented increases in hepatic and steatosis in HFD-fed mice. The opposite was observed when miR-291b-3p was overexpressed in the livers of chow-fed C57BL/6J wild-type mice. In vitro studies revealed that silencing of miR-291b-3p in NCTC1469 hepatic cells ameliorated oleic / mixture-induced elevation of cellular triglycerides. Importantly, we identified AMP-activated protein kinase (AMPK)-α1 as a direct target of miR-291b-3p. Using metformin, an activator of AMPK, we showed that AMPK activation-induced inhibition of hepatic lipid accumulation was accompanied by reduced expression of miR-291b-3p in the liver. Liver miR-291b-3p promoted hepatic and lipid accumulation in mice. AMPKα1 is a direct target of miR-291b-3p. In conclusion, our findings indicate that miR-291b-3p promotes hepatic by suppressing AMPKα1 expression and activity, indicating the therapeutic potential of miR-291b-3p inhibitors in fatty liver disease.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: lipogenesis

Dimethylarginine Dimethylaminohydrolase 1 Protects Against High-Fat Diet-Induced Hepatic Steatosis and Insulin Resistance in Mice.

High plasma concentrations of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, are associated with hepatic dysfunction in patients with nonalcoholic fatty liver disease (NAFLD). However, it is unknown whether ADMA is involved in the pathogenesis of NAFLD. Dimethylarginine dimethylaminohydrolase 1 (DDAH1) is an enzyme that degrades ADMA. In this study, we used Ddah1 mice to investigate the effects of the ADMA/DDAH1 pathway on high-fat diet (HFD)-induced hepatic steatosis.After HFD feeding for 20 weeks, Ddah1 mice were more obese and had developed more severe hepatic steatosis and worse insulin resistance compared with wild-type (WT) mice. In the livers of HFD-fed mice, loss of DDAH1 resulted in higher levels of lipogenic genes, lower expression of β-oxidation genes, and greater induction of oxidative stress, endoplasmic reticulum stress, and inflammation than in the WT livers. Furthermore, ADMA treatment in HepG2 cells led to oxidative stress and steatosis, whereas overexpression of DDAH1 attenuated -induced steatosis, oxidative stress, and inflammation. Innovation and Conclusion: Our results provide the first direct evidence that the ADMA/DDAH1 pathway has a marked effect on hepatic and steatosis induced by HFD feeding. Our findings suggest that strategies to increase DDAH1 activity in hepatocytes may provide a novel approach to attenuate NAFLD development. Antioxid. Redox Signal. 26, 598-609.

Keyword: lipogenesis

High α-tocopherol dosing increases lipid metabolism by changing redox state in damaged rat gastric mucosa and liver after ethanol treatment.

Regeneration of ethanol-injured rat gastric mucosa must undergo changes in major metabolic pathways to achieve DNA replication and cell proliferation. These events are highly dependent on glucose utilization and inhibited by vitamin E (VE) (α-tocopherol) administration. Therefore, the present study aimed at assessing lipid metabolism in the gastric mucosa and ethanol-induced gastric damage and the effect of α-tocopherol administration. For this, rates of fatty β-oxidation and were tested in gastric mucosa samples. Through histological analysis, we found loss of the mucosa\'s superficial epithelium, which became gradually normalized during the recovery period. Proliferation of gastric mucosa occurred with augmented formation of β-oxidation by-products, diminished synthesis of triacylglycerols (TGs), as well as of phospholipids, and a reduced cytoplasmic NAD/NADH ratio, whereas the mitochondrial redox NAD/NADH ratio was much less affected. In addition, α-tocopherol increased utilization in the gastric mucosa, which was accompanied by the induction of \'mirror image\' effects on the cell redox state, reflected in an inhibited cell gastric mucosa proliferation by the vitamin administration. In conclusion, the present study shows, for the first time, the role of lipid metabolism in the adaptive cell gastric mucosa changes that drive proliferation after a chronic insult. Moreover, α-tocopherol increased gastric mucosa utilization of associated with energy production. These events could be associated with its antioxidant properties in co-ordination with regulation of genes and cell pathways, including changes in the cell NAD/NADH redox state.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: lipogenesis

Decreased lipid metabolism but increased FA biosynthesis are coupled with changes in liver microRNAs in obese subjects with NAFLD.

Many controversies regarding the association of liver miRNAs with obesity and nonalcoholic fatty liver diseases (NAFLD) call for additional validations. This study sought to investigate variations in genes and hepatic miRNAs in a sample of obese patients with or without NAFLD and human hepatocytes (HH).A total of 60 non-consecutive obese women following bariatric surgery were recruited. Subjects were classified as NAFLD (n=17), borderline (n=24) and controls (n=19) with normal enzymatic profile, liver histology and ultrasound assessments. Profiling of 744 miRNAs was performed in 8 obese women with no sign of hepatic disease and 11 NAFLD patients. Additional validation and expression of genes related to de novo fatty (FA) biosynthesis, uptake, transport and β-oxidation; glucose metabolism, and inflammation was tested in the extended sample. Induction of NAFLD-related genes and miRNAs was examined in HepG2 cells and primary HH treated with (PA), a combination of palmitate and oleic , or high glucose, and insulin (HG) mimicking insulin resistance in NAFLD.In the discovery sample, 14 miRNAs were associated with NAFLD. Analyses in the extended sample confirmed decreased miR-139-5p, miR-30b-5p, miR-122-5p and miR-422a, and increased miR-146b-5p in obese subjects with NAFLD. Multiple linear regression analyses disclosed that NAFLD contributed independently to explain miR-139-5p (P=0.005), miR-30b-5p (P=0.005), miR-122-5p (P=0.021), miR-422a (P=0.007) and miR-146a (P=0.033) expression variance after controlling for confounders. Decreased miR-122-5p in liver was associated with impaired FA usage. Expression of inflammatory and macrophage-related genes was opposite to decreased miR-30b-5p, miR-139-5p and miR-422a, whereas increased miR-146b-5p was associated with FABP4 and decreased glucose metabolism and FA mobilization. In partial agreement, PA (but not HG) led to decreased miR-139-5p, miR-30b-5p, miR-422a and miR-146a in vitro, in parallel with increased and FA transport, decreased glucose metabolism and diminished FA oxidation.This study confirms decreased liver glucose and lipid metabolism but increased FA biosynthesis coupled with changes in five unique miRNAs in obese patients with NAFLD.

Keyword: lipogenesis

Dose- and type-dependent effects of long-chain fatty acids on and of bovine adipocytes.

Differentiation and lipid metabolism of adipocytes have a great influence on milk performance, health, and feed efficiency of dairy cows. The effects of dietary long-chain fatty acids (FA) on and of dairy cows are often confounded by other nutritional and physiological factors in vivo. Therefore, this study used an in vitro approach to study the effect of dose and type of long-chain FA on and of bovine adipocytes. Stromal vascular cells were isolated from adipose tissue of dairy cows and induced into mature adipocytes in the presence of various long-chain FA including myristic, , stearic, oleic, or linoleic . When concentrations of myristic, , and oleic acids in adipogenic mediums were 150 and 200 μM, the induced mature adipocytes had greater lipid content compared with other concentrations of FA. In addition, mature adipocytes induced at 100 μM stearic and 300 μM linoleic had the greatest content of lipid than at other concentrations. High concentrations of saturated FA were more toxic for cells than the same concentration of unsaturated FA during the induction. When commitment stage was solely treated with FA, the number of differentiated mature adipocytes was greater for oleic and linoleic acids than other FA. When the maturation stage was treated with FA, the number of mature adipocytes was not affected, but the lipid content in adipocytes was affected and ranked oleic > linoleic > myristic > stearic > . In summary, this study showed that and of bovine adipocytes were differentially affected by long-chain FA, with unsaturated FA more effective than saturated FA.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Microalgal lipids biochemistry and biotechnological perspectives.

In the last few years, there has been an intense interest in using microalgal lipids in food, chemical and pharmaceutical industries and cosmetology, while a noteworthy research has been performed focusing on all aspects of microalgal lipid production. This includes basic research on the pathways of solar energy conversion and on lipid biosynthesis and catabolism, and applied research dealing with the various biological and technical bottlenecks of the lipid production process. In here, we review the current knowledge in microalgal lipids with respect to their metabolism and various biotechnological applications, and we discuss potential future perspectives. The committing step in fatty biosynthesis is the carboxylation of acetyl-CoA to form malonyl-CoA that is then introduced in the fatty synthesis cycle leading to the formation of and stearic acids. Oleic may also be synthesized after stearic desaturation while further conversions of the fatty acids (i.e. desaturations, elongations) occur after their esterification with structural lipids of both plastids and the endoplasmic reticulum. The aliphatic chains are also used as building blocks for structuring storage acylglycerols via the Kennedy pathway. Current research, aiming to enhance in the microalgal cell, is focusing on over-expressing key-enzymes involved in the earlier steps of the pathway of fatty synthesis. A complementary plan would be the repression of lipid catabolism by down-regulating acylglycerol hydrolysis and/or β-oxidation. The tendency of oleaginous microalgae to synthesize, apart from lipids, significant amounts of other energy-rich compounds such as sugars, in processes competitive to , deserves attention since the lipid yield may be considerably increased by blocking competitive metabolic pathways. The majority of microalgal production occurs in outdoor cultivation and for this reason biotechnological applications face some difficulties. Therefore, algal production systems need to be improved and harvesting systems need to be more effective in order for their industrial applications to become more competitive and economically viable. Besides, a reduction of the production cost of microalgal lipids can be achieved by combining lipid production with other commercial applications. The combined production of bioactive products and lipids, when possible, can support the commercial viability of both processes. Hydrophobic compounds can be extracted simultaneously with lipids and then purified, while hydrophilic compounds such as proteins and sugars may be extracted from the defatted biomass. The microalgae also have applications in environmental biotechnology since they can be used for bioremediation of wastewater and to monitor environmental toxicants. Algal biomass produced during wastewater treatment may be further valorized in the biofuel manufacture. It is anticipated that the high microalgal lipid potential will force research towards finding effective ways to manipulate biochemical pathways involved in lipid biosynthesis and towards cost effective algal cultivation and harvesting systems, as well.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

GLUT4 Expression in Adipocytes Regulates De Novo and Levels of a Novel Class of Lipids With Antidiabetic and Anti-inflammatory Effects.

Adipose tissue (AT) regulates systemic insulin sensitivity through multiple mechanisms, and alterations in de novo appear to contribute. Mice overexpressing GLUT4 in adipocytes (AG4OX) have elevated AT and enhanced glucose tolerance despite being obese and having elevated circulating fatty acids. Lipidomic analysis of AT identified a structurally unique class of lipids, branched fatty esters of hydroxy-fatty acids (FAHFAs), which were elevated in AT and serum of AG4OX mice. esters of hydroxy-stearic acids (PAHSAs) are among the most upregulated FAHFA families in AG4OX mice. Eight PAHSA isomers are present in mouse and human tissues. PAHSA levels are reduced in insulin resistant people, and levels correlate highly with insulin sensitivity. PAHSAs have beneficial metabolic effects. Treatment of obese mice with PAHSAs lowers glycemia and improves glucose tolerance while stimulating glucagon-like peptide 1 and insulin secretion. PAHSAs also reduce inflammatory cytokine production from immune cells and ameliorate adipose inflammation in obesity. PAHSA isomer concentrations are altered in physiological and pathophysiological conditions in a tissue- and isomer-specific manner. The mechanisms most likely involve changes in PAHSA biosynthesis, degradation, and secretion. The discovery of PAHSAs reveals the existence of previously unknown endogenous lipids and biochemical pathways involved in metabolism and inflammation, two fundamental physiological processes.© 2016 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

Keyword: lipogenesis

Associations of erythrocyte fatty acids in the de novo pathway with risk of metabolic syndrome in a cohort study of middle-aged and older Chinese.

Experimental studies suggest that elevated de novo (DNL) might be involved in the pathogenesis of metabolic disorders. Few prospective studies have been conducted, especially among populations with a high carbohydrate intake, to determine whether DNL fatty acids are associated with the risk of the metabolic syndrome (MetS).We aimed to investigate associations of erythrocyte fatty acids in the DNL pathway-including myristic (14:0), (16:0), palmitoleic (16:1n-7), hexadecenoic (16:1n-9), stearic (18:0), vaccenic (18:1n-7), and oleic (18:1n-9)-with the risk of MetS in a Chinese population with an average carbohydrate intake of >60% of energy.A total of 1176 free-living Chinese men and women aged 50-70 y from Beijing and Shanghai were included in our analysis, giving rise to 412 incident MetS cases during 6 y of follow-up. Erythrocyte fatty acids and metabolic traits were measured in these participants.Erythrocyte fatty acids in the DNL pathway were correlated with a high ratio of carbohydrate-to-fat intake, less favorable lipid profiles, and elevated liver enzymes at baseline. In comparison with the lowest quartile, RRs (95% CIs) of MetS in the highest quartile were 1.30 (1.04, 1.62; P-trend = 0.007) for 16:1n-7, 1.48 (1.17, 1.86; P-trend < 0.001) for 16:1n-9, 1.26 (1.01, 1.56; P-trend = 0.06) for 18:1n-7, and 1.51 (1.19, 1.92; P-trend < 0.001) for 18:1n-9 after multivariate adjustment for lifestyle factors and body mass index. Moreover, 16:0 and 16:1n-7 were associated with an elevated risk of diabetes.Our findings suggest that fatty acids in the DNL pathway are independently associated with an elevated risk of metabolic disorders.

Keyword: lipogenesis

Quercetin induces HepG2 cell apoptosis by inhibiting fatty biosynthesis.

Quercetin can inhibit the growth of cancer cells with the ability to act as a \'chemopreventer\'. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell-cycle arrest and/or apoptosis, as well as its antioxidant functions. Quercetin can also reduce . Previous studies have shown that quercetin has potent inhibitory effects on animal fatty synthase (FASN). In the present study, activity of quercetin was evaluated in human liver cancer HepG2 cells. Intracellular FASN activity was calculated by measuring the absorption of NADPH via a spectrophotometer. MTT assay was used to test the cell viability, immunoblot analysis was performed to detect FASN expression levels and the apoptotic effect was detected by Hoechst 33258 staining. In the present study, it was found that quercetin could induce apoptosis in human liver cancer HepG2 cells with overexpression of FASN. This apoptosis was accompanied by the reduction of intracellular FASN activity and could be rescued by 25 or 50 μM exogenous acids, the final product of FASN-catalyzed synthesis. These results suggested that the apoptosis induced by quercetin was via the inhibition of FASN. These findings suggested that quercetin may be useful for preventing human liver cancer.

Keyword: lipogenesis

Chronic administration of saturated fats and fructose differently affect SREBP activity resulting in different modulation of Nrf2 and Nlrp3 inflammasome pathways in mice liver.

The overconsumption of both saturated fats and fructose in the modern society has been related to the development of nonalcoholic fatty liver disease (NAFLD). However, the specific contribution of individual dietary components on the progression of NAFLD to nonalcoholic steatohepatitis (NASH) has been poorly investigated. Therefore, the aim of our study was to investigate the dissimilar effects of these two dietary components on selected proinflammatory and antioxidant pathways in the liver of C57BL/6 mice fed a standard (SD), a 45% saturated fat (HFAT) or a 60% fructose (HFRT) diet for 12 weeks. HFAT diet evoked systemic metabolic alterations and overweight, not observed in HFRT mice. However, HFRT mice had a greater hepatic triglyceride deposition with increased ratio of triacylglycerols containing the compared to HFAT, as assessed by liquid chromatography-mass spectrometry analysis. This effect is due to the higher activation of the SCAP/SREBP1c lipogenic pathway by HFRT feeding. In addition, we found inhibition of Keap1/Nrf2 antioxidant signaling and more robust stimulation of the Nlrp3 inflammasome pathway in the livers of HFRT-fed mice when compared with HFAT-fed mice, which is consistent with the recent finding that palmitate and SREBP1c are implicated in hepatic oxidative stress and inflammation. These effects were associated with increased hepatic inflammation, as confirmed by high expression of markers of leukocyte infiltration in the HFRT group. Thus, we hypothesize an amplifying loop among , palmitate, Nrf2 and Nlrp3 that leads to a higher risk of NAFLD progression to NASH in a high-fructose diet compared to a high-saturated fat intake.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells.

Acetyl-CoA carboxylases (ACC) 1 and 2 catalyze the carboxylation of acetyl-CoA to malonyl-CoA and depend on biotin as a coenzyme. ACC1 localizes in the cytoplasm and produces malonyl-CoA for fatty (FA) synthesis. ACC2 localizes in the outer mitochondrial membrane and produces malonyl-CoA that inhibits FA import into mitochondria for subsequent oxidation. We hypothesized that ACCs are checkpoints in adipocyte differentiation and tested this hypothesis using the ACC1 and ACC2 inhibitor soraphen A (SA) in murine 3T3-L1 preadipocytes. When 3T3-L1 cells were treated with 100nM SA for 8 days after induction of differentiation, the expression of PPARγ mRNA and FABP4 mRNA decreased by 40% and 50%, respectively, compared with solvent controls; the decrease in gene expression was accompanied by a decrease in FABP4 protein expression and associated with a decrease in lipid droplet accumulation. The rate of FA oxidation was 300% greater in SA-treated cells compared with vehicle controls. Treatment with exogenous palmitate restored PPARγ and FABP4 mRNA expression and FABP4 protein expression in SA-treated cells. In contrast, SA did not alter lipid accumulation if treatment was initiated on day eight after induction of differentiation. We conclude that loss of ACC1-dependent FA synthesis and loss of ACC2-dependent inhibition of FA oxidation prevent lipid accumulation in adipocytes and inhibit early stages of adipocyte differentiation.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Altered lipid metabolism in residual white adipose tissues of Bscl2 deficient mice.

Mutations in BSCL2 underlie human congenital generalized lipodystrophy type 2 disease. We previously reported that Bscl2 (-/-) mice develop lipodystrophy of white adipose tissue (WAT) due to unbridled lipolysis. The residual epididymal WAT (EWAT) displays a browning phenotype with much smaller lipid droplets (LD) and higher expression of brown adipose tissue marker proteins. Here we used targeted lipidomics and gene expression profiling to analyze lipid profiles as well as genes involved in lipid metabolism in WAT of wild-type and Bscl2(-/-) mice. Analysis of total saponified fatty acids revealed that the residual EWAT of Bscl2(-/-) mice contained a much higher proportion of oleic 18:1n9 concomitant with a lower proportion of 16:0 , as well as increased n3- polyunsaturated fatty acids (PUFA) remodeling. The acyl chains in major species of triacylglyceride (TG) and diacylglyceride (DG) in the residual EWAT of Bscl2(-/-) mice were also enriched with dietary fatty acids. These changes could be reflected by upregulation of several fatty elongases and desaturases. Meanwhile, Bscl2(-/-) adipocytes from EWAT had increased gene expression in lipid uptake and TG synthesis but not de novo . Both mitochondria and peroxisomal β-oxidation genes were also markedly increased in Bscl2(-/-) adipocytes, highlighting that these machineries were accelerated to shunt the lipolysis liberated fatty acids through uncoupling to dissipate energy. The residual subcutaneous white adipose tissue (ScWAT) was not browning but displays similar changes in lipid metabolism. Overall, our data emphasize that, other than being essential for adipocyte differentiation, Bscl2 is also important in fatty remodeling and energy homeostasis.

Keyword: lipogenesis

Preservation of high-fat diet-induced femoral trabecular bone loss through genetic target of TNF-α.

Obesity and osteoporosis are two common chronic diseases, however, the basis for the correlation between them remains largely unknown. The pro-inflammation cytokine tumor necrosis factor-alpha (TNF-α) plays important roles in lipid and bone metabolisms, which may be a good candidate in the correlation between obesity and osteoporosis. We investigated the pathological roles of TNF-α in high-fat diet (HFD)-induced bone loss. Wild-type (WT) mice and TNF-α knockout (TNF-α(-/-)) mice were fed with the standard diet or the HFD for 12 weeks. Bone marrow stromal cells (BMSCs) from both genotypes were induced to differentiate into osteoblasts and treated with (PA). Bone mass and microstructure of femurs were evaluated by micro-CT. Lipid and bone metabolisms were investigated by histological and plasma analyses, and real-time PCR. On the HFD, both TNF-α(-/-) and WT mice presented notable visceral obesity, dyslipidemia. and osteoclastogenesis were enhanced, while osteoblastogenesis was reduced in both genotypes. However, the changes were significantly different between TNF-α(-/-) and WT mice after the HFD. The gain of body and fat-pad weight was less and adipocyte area was smaller by 22 % in TNF-α(-/-) mice. Osteoclast numbers and plasma CTX level were lower by 40 % and by 23 % in TNF-α(-/-) mice. There were more ALP positive cells in the PA-treated TNF-α(-/-) BMSCs. mRNA expression of PPAR-γ was lower while that of Runx2 was higher in the bone from TNF-α(-/-) HFD group and in the PA-treated TNF-α(-/-) BMSCs, compared to WT on the same treatment. Furthermore, femoral trabecular bone mass and trabecular bone number were significantly decreased in WT mice on the HFD, whereas they were increased by 1.56-fold and 1.53-fold, respectively, in TNF-α(-/-) mice on the same diet (P < 0.05). Our results demonstrated that TNF-α gene knockout retained HFD-induced femoral trabecular bone loss mainly by suppressing and osteoclastogenesis, and enhancing osteoblastogenesis, which suggests that TNF-α plays a critical role in the development of HFD-related bone metabolic disorders and it may be a new potential therapeutic target for obesity-related bone loss.

Keyword: lipogenesis

Lipopolysaccharide binding protein is an adipokine involved in the resilience of the mouse adipocyte to inflammation.

Lipopolysaccharide (LPS) binding protein (LBP) is a novel 65 kDa adipokine, linked to adipose tissue (AT) inflammation, obesity and insulin resistance, that inhibits adipocyte differentiation. Here, we investigated the molecular mechanisms behind these detrimental effects on through whole-genome transcriptomics and in vitro experiments.Permanent and transient knockdown (KD) and co-culture experiments were performed in 3T3-L1 and 3T3-F442A cell lines during adipocyte differentiation. Microarray gene expression was performed using Genechip Affymetrix technology and validated by real-time PCR.LBP KD of 3T3-L1 cells led to a potentiated adipocyte differentiation with a dose-response relationship; genes involved in mitochondrial biogenesis, fatty metabolism and peroxisome proliferator-activated receptor γ (PPAR-γ) action were dramatically upregulated in parallel to increased insulin signalling. Cells with LBP KD became refractory to proinflammatory cytokines and other inflammatory stimuli (LPS and palmitate). This phenotype, mediated through disrupted nuclear factor κB (NFκB) signalling, was reversed by a soluble factor present in a co-culture with native cells and by exogenous LBP. Double-silencing of LBP and toll-like receptor 4 (TLR4) again rendered these cells insensitive to co-culture, LBP and inflammatory factors.In summary, LBP is a proinflammatory soluble adipokine that acts as a brake for , strengthening the negative effects of palmitate and LPS on adipocyte differentiation.

Keyword: lipogenesis

Chalcones suppress fatty -induced lipid accumulation through a LKB1/AMPK signaling pathway in HepG2 cells.

Excessive lipid accumulation in the liver has been proposed to cause hyperlipidemia, diabetes and fatty liver disease. 4-Hydroxyderricin (4HD), xanthoangelol (XAG), cardamonin (CAR) and flavokawain B (FKB) are chalcones that have exhibited various biological effects against obesity, inflammation, and diabetes; however, little is known about the inhibitory effects of these chalcones on fatty liver disease. In the present study, we investigated the ability of 4HD, XAG, CAR, and FKB to reduce lipid accumulation in hepatocytes. When HepG2 cells were treated with a mixture of fatty acids (FAs; : oleic = 1 : 2 ratio), significant lipid accumulation was observed. Under the same experimental conditions, addition of chalcones at 5 μM significantly suppressed the FA-induced lipid accumulation. We found that the expression of sterol regulatory element-binding protein-1 (SREBP-1), a key molecule involved in , was decreased in these chalcone-treated cells. We also found that these chalcones increased the expression of peroxisome proliferator-activated receptor α (PPARα), which is involved in FA oxidation. Moreover, these chalcones increased phosphorylation of AMP-activated protein kinase (AMPK) and liver kinase B1 (LKB1), upstream regulators of SREBP-1 and PPARα. We confirmed that an AMPK inhibitor, compound C, reversed chalcone-induced changes in SREBP-1 and PPARα expression in the HepG2 cells. Collectively, we found that 4HD, XAG, CAR, and XAG attenuated lipid accumulation through activation of the LKB1/AMPK signaling pathway in HepG2 cells.

Keyword: lipogenesis

The activation of peroxisome proliferator-activated receptor γ is regulated by Krüppel-like transcription factors 6 & 9 under steatotic conditions.

Liver steatosis is characterised by lipid droplet deposition in hepatocytes that can leads to an inflammatory and fibrotic phenotype. Peroxisome proliferator-activated receptors (PPARs) play key roles in energetic homeostasis by regulating lipid metabolism in hepatic tissue. In adipose tissue PPARγ regulates the adipocyte differentiation by promoting the expression of lipid-associated genes. Within the liver PPARγ is up-regulated under steatotic conditions; however, which transcription factors participate in its expression is not completely understood. Krüppel-like transcription factors (KLFs) regulate various cellular mechanisms, such as cell proliferation and differentiation. KLFs are key components of by regulating the expression of PPARγ and other proteins such as the C-terminal enhancer binding protein (C/EBP). Here, we demonstrate that the transcript levels of Klf6, Klf9 and Pparγ are increased in response to a steatotic insult in vitro. Chromatin immunoprecipitation (ChIp) experiments showed that klf6 and klf9 are actively recruited to the Pparγ promoter region under these conditions. Accordingly, the loss-of-function experiments reduced cytoplasmic triglyceride accumulation. Here, we demonstrated that KLF6 and KLF9 proteins directly regulate PPARγ expression under steatotic conditions.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Risk of diabetes associated with fatty acids in the de novo pathway is independent of insulin sensitivity and response: the Insulin Resistance Atherosclerosis Study (IRAS).

To examine the associations of fatty acids in the de novo (DNL) pathway, specifically myristic (14:0), (16:0), palmitoleic (c16:1\u2009n-7), myristoleic (c14:1n5), stearic (18:0) and oleic (c18:1\u2009n-9), with 5-year risk of type 2 diabetes. We hypothesized that DNL fatty acids are associated with risk of type 2 diabetes independent of insulin sensitivity.We evaluated 719 (mean age 55.1±8.5 years, 44.2% men, 42.3% Caucasians) participants from the Insulin Resistance Atherosclerosis Study. Multivariable logistic regression models with and without adjustment of insulin sensitivity were used to assess prospective associations of DNL fatty acids with incident type 2 diabetes.Type 2 diabetes incidence was 20.3% over 5\u2009years. In multivariable regression models, , palmitoleic, myristic, myristoleic and oleic acids were associated with increased risk of type 2 diabetes (p<0.05). had the strongest association (OR per standard unit of 1.46; 95%\u2009CI 1.23 to 1.76; p<0.001), which remained similar with addition of insulin sensitivity and acute insulin response (AIR) to the model (OR 1.36; 95%\u2009CI 1.09 to 1.70, p=0.01). Oleic and palmitoleic acids were also independently associated with incident type 2 diabetes. In multivariable models, ratios of fatty acids corresponding to stearoyl CoA desaturase-1 and Elovl6 enzymatic activity were significantly associated with risk of type 2 diabetes independent of insulin sensitivity and AIR.We observed associations of DNL fatty acids with type 2 diabetes incidence independent of insulin sensitivity.

Keyword: lipogenesis

De novo biosynthesis of fatty acids from α-D-glucose in parasitoid wasps of the Nasonia group.

Fatty acids are indispensable primary metabolites for virtually any organism on earth and thus enzymatic machinery enabling de novo production of fatty acids from carbohydrates is highly conserved. A series of studies has questioned the ubiquity of in parasitoid wasps suggesting that the vast majority of species have lost the ability to synthesize fatty acids de novo. One such species is Nasonia vitripennis, which, like the congeneric species N. giraulti and N. longicornis, uses a fatty -derived male sex pheromone for sexual communication. Here we demonstrate by feeding fully C-labeled α-D-glucose and analyzing insect-derived fatty methyl esters and the male sex pheromone by coupled gas chromatography/mass spectrometry that both males and females of N. vitripennis as well as N. giraulti and N. longicornis are capable of synthesizing fatty acids de novo. We furthermore show by a proteomics approach that predicted fatty synthase, ATP-citrate synthase, and acetyl-CoA carboxylase, key enzymes of , are expressed in the male pheromone gland of N. vitripennis and N. giraulti. Labeling experiments with Urolepis rufipes, a closely related species producing a male sex pheromone independently of fatty acids via the mevalonate pathway, revealed that both sexes are likewise able to synthesize fatty acids de novo. We conclude that the parasitoid wasp species studied here, irrespective of the biosynthetic origin of their sex pheromones, are capable of responding flexibly to lipid shortage during their adult life by keeping enzymatic machinery for running.Copyright © 2019. Published by Elsevier Ltd.

Keyword: lipogenesis

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low fat diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic lipid accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and diabetes, and (3) dietary SFA type impacts LCPUFA metabolism.Published by Elsevier Inc.

Keyword: lipogenesis

miR-192-5p regulates lipid synthesis in non-alcoholic fatty liver disease through SCD-1.

To evaluate the levels of miR-192-5p in non-alcoholic fatty liver disease (NAFLD) models and demonstrate the role of miR-192-5p in lipid accumulation.Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet (HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic miR-192-5p and stearoyl-CoA desaturase 1 (SCD-1) levels were measured. MiR-192-5p mimic and inhibitor and SCD-1 siRNA were transfected into Huh7 cells exposed to (PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic miR-192-5p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection ( < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of miR-192-5p and SCD-1 protein levels, respectively ( < 0.01). Transfection with miR-192-5p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively ( < 0.01). Luciferase activity was suppressed and enhanced by miR-192-5p mimic and inhibitor, respectively, in wild-type SCD-1 ( < 0.01) but not in mutant SCD-1. MiR-192-5p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 siRNA transfection abrogated the lipid deposition aggravated by miR-192-5p inhibitor ( < 0.01).This study demonstrates that miR-192-5p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.

Keyword: lipogenesis

Depot-specific differences in fatty composition and distinct associations with lipogenic gene expression in abdominal adipose tissue of obese women.

Cardiometabolic diseases are primarily linked to enlarged visceral adipose tissue (VAT). However, some data suggest heterogeneity within the subcutaneous adipose tissue (SAT) depot with potential metabolic differences between the superficial SAT (sSAT) and deep SAT (dSAT) compartments. We aimed to investigate the heterogeneity of these three depots with regard to fatty (FA) composition and gene expression. Adipose tissue biopsies were collected from 75 obese women undergoing laparoscopic gastric bypass surgery. FA composition and gene expression were determined with gas chromatography and quantitative real-time-PCR, respectively. Stearoyl CoA desaturase-1 (SCD-1) activity was estimated by product-to-precursor FA ratios. All polyunsaturated FAs (PUFA) with 20 carbons were consistently lower in VAT than either SAT depots, whereas essential PUFA (linoleic , 18:2n-6 and α-linolenic , 18:3n-3) were similar between all three depots. Lauric and were higher and lower in VAT, respectively. The SCD-1 product palmitoleic as well as estimated SCD-1 activity was higher in VAT than SAT. Overall, there was a distinct association pattern between lipid metabolizing genes and individual FAs in VAT. In conclusion, SAT and VAT are two distinct depots with regard to FA composition and expression of key lipogenic genes. However, the small differences between sSAT and dSAT suggest that FA metabolism of SAT is rather homogenous.

Keyword: lipogenesis

Acanthoic modulates in nonalcoholic fatty liver disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic fatty liver disease (NAFLD) associated with lipid accumulation by activating Farnesoid X receptor (FXR) and liver X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high-fat (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on lipid accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, liver index, and liver lipid droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated lipid accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating lipid accumulation and fatty synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Desaturation index versus isotopically measured de novo as an indicator of acute systemic .

High carbohydrate feeding is known to increase plasma triglycerides as well as hepatic de novo (DNL) and may be implicated in the development of hepatic insulin resistance and fatty liver. Unfortunately, it is technically challenging to determine what proportion of circulating plasma triglycerides have been derived from the newly synthesized fatty acids in the postprandial period. The aims of this study were to 1) characterize the changes in the plasma postprandial total fatty pool in beagles following the consumption of meals containing 44% (Control) and 74% (High Sucrose) carbohydrate and 2) determine if changes in plasma fatty concentration and delta-9 desaturation index (DI) would be useful as simple and easy to measure biomarkers of systemic DNL.No differences in plasma total (16:0), stearic (18:0) and oleic (18:1) concentrations or delta-9 DI for the total 18:0 and 18:1 pools between High Sucrose and Controls were observed. However, newly synthesized 16:0 (2.6 ± 0.2% vs. 8.8 ± 2.0%; p = 0.016), 18:0 (0.93 ± 0.2% vs. 4.1 ± 1.7%; p = 0.007) and 18:1 (0.29 ± 0.09% vs. 3.5 ± 1.2%; p = 0.017) were higher in High Sucrose versus Control animals, respectively. Also, the delta-9 DI for the newly synthesized 18:0 and 18:1 pools was higher at 2 and 6 hours postprandial, with a pattern of change which supports the increased stearoyl-CoA desaturase (SCD-1) activity following high carbohydrate feeding followed by a down regulation of this enzyme.Our data show that high sucrose meals increase the relative contribution of systemic DNL produced fatty acids to the total postprandial plasma fatty pool. These data also show that a different pattern of both fatty synthesis and disposal occurs depending on energy and macronutrient profile of the meal. These changes are in spite of no observable changes in the plasma concentrations or ratios of the total fatty pool opposed to the observed changes in the newly synthesized fatty pool.

Keyword: lipogenesis

Discovery of a class of endogenous mammalian lipids with anti-diabetic and anti-inflammatory effects.

Increased adipose tissue is associated with enhanced insulin sensitivity. Mice overexpressing the Glut4 glucose transporter in adipocytes have elevated and increased glucose tolerance despite being obese with elevated circulating fatty acids. Lipidomic analysis of adipose tissue revealed the existence of branched fatty esters of hydroxy fatty acids (FAHFAs) that were elevated 16- to 18-fold in these mice. FAHFA isomers differ by the branched ester position on the hydroxy fatty (e.g., -9-hydroxy-stearic-, 9-PAHSA). PAHSAs are synthesized in vivo and regulated by fasting and high-fat feeding. PAHSA levels correlate highly with insulin sensitivity and are reduced in adipose tissue and serum of insulin-resistant humans. PAHSA administration in mice lowers ambient glycemia and improves glucose tolerance while stimulating GLP-1 and insulin secretion. PAHSAs also reduce adipose tissue inflammation. In adipocytes, PAHSAs signal through GPR120 to enhance insulin-stimulated glucose uptake. Thus, FAHFAs are endogenous lipids with the potential to treat type 2 diabetes.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

MicroRNA-199a-3p attenuates hepatic by targeting Sp1.

Emerging studies have demonstrated that microRNAs (miRs) are profoundly involved in non-alcoholic fatty liver disease (NAFLD) and related metabolic diseases. Previously, we revealed a repertoire of miRs dysregulated in NAFLD by high-throughput sequencing. Here, we showed that microRNA-199a-3p was down-regulated in the livers of C57BL/6J mice fed a high-fat-diet (HFD) and oleic /-induced Hepa1-6 cells. Gain-of-function and loss-of-function studies demonstrated that microRNA-199a-3p exhibited a suppressive role in hepatic . Adenoviral mediated microRNA-199a-3p expression in C57BL/6J mice largely attenuated triglyceride (TG) accumulation and expression of lipogenic genes. Furthermore, we identified Specificity Protein 1 (Sp1) as the functional target of miR-124. Restoration of Sp1 expression largely compromised the effect of microRNA-199a-3p on hepatic TG metabolism. Taken together, our findings uncover a novel function of microRNA-199a-3p/Sp1 axis in NAFLD and provide a mechanism underlying perturbations of hepatic TG homeostasis.

Keyword: lipogenesis

Synthesis and biological evaluations of marine oxohexadecenoic acids: PPARα/γ dual agonism and anti-diabetic target gene effects.

Obesity and associated disorders such as metabolic syndrome and type 2 diabetes (T2D) have reached epidemic proportions. Several natural products have been reported as Peroxisome Proliferator-Activated Receptor (PPAR) agonists, functioning as lead compounds towards developing new anti-diabetic drugs due to adverse side effects of existing PPAR drugs. We recently isolated and identified (7E)-9-oxohexadec-7-enoic (1) and (10E)-9-oxohexadec-10-enoic (2) from the marine algae Chaetoceros karianus. Herein we report the total synthesis, pharmacological characterization, and biological evaluations of these naturally occurring oxo-fatty acids (oFAs). The syntheses of 1 and 2 afforded sufficient material for extensive biological evaluations. Both oFAs show an appreciable dose-dependent activation of PPARα and -γ, with EC values in the micromolar range, and an ability to regulate important PPAR target genes in hepatocytes and adipocytes. Moreover, both 1 and 2 are able to drive when evaluated in the Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocyte cell model, but with lowered expression of adipocyte markers and reduced lipid accumulation compared to the drug rosiglitazone. This seems to be caused by a transient upregulation of PPARγ and C/EBPα expression. Importantly, whole transcriptome analysis shows that both compounds induce anti-diabetic gene programs in adipocytes by upregulating insulin-sensitizing adipokines and repressing pro-inflammatory cytokines.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: lipogenesis

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid Metabolism, Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free fatty (FFA)- and oleic -and high-fat diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice liver were measured, and the expression levels of genes associated with , fatty oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of -related genes, but upregulated the expression of genes associated with fatty oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of lipid metabolism, inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: lipogenesis

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the liver was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and obesity in male mice, whereas their metabolites inhibit and lipid uptake in vitro.

Keyword: lipogenesis

Palmitoleic (16:1 cis-9) and cis-vaccenic (18:1 cis-11) alter in bovine adipocyte cultures.

Our objectives were to: (1) confirm elongation products of palmitoleic (16:1 cis-9) elongation in vitro using stable isotopes and (2) evaluate if exogenous supplementation of palmitoleic , elongation products, or both are responsible for decreased desaturation and rates observed with palmitoleic supplementation in bovine adipocytes. Stromal vascular cultures were isolated from adipose tissue of two beef carcasses, allowed to reach confluence, held for 2 days, and differentiated with a standard hormone cocktail (day 0). On day 2, secondary differentiation media containing 1 of 4 fatty treatments [0 μM fatty (control), or 150 μM (16:0), palmitoleic, or cis-vaccenic (18:1 cis-11)] was added for 4 days. On day 6, cells were incubated with [(13)C] 16:1, [(13)C] 2, or [(13)C] 18:0 to estimate elongation, lipogenic, and desaturation rates using gas chromatography-mass spectrometry. Enrichment of [(13)C] 18:1 cis-11 confirmed 18:1 cis-11 is an elongation product of 16:1. Additionally, [(13)C] label was seen in 20:1 cis-13 and cis-9, cis-11 CLA. Synthesis of [(13)C] 16:0 from [(13)C] 2 was reduced (P < 0.05) in palmitoleic and cis-vaccenic -treated compared with control cells following 36 h incubation. By 12 h of [(13)C] 18:0 incubation, cells supplemented with palmitoleic had reduced (P < 0.05) [(13)C] 18:1 cis-9 compared with all other treatments. Gene expression and fatty results support isotopic data for and desaturation. Therefore, palmitoleic is actively elongated in vitro and its elongation product, cis-vaccenic , can also reduce . However, inhibition of desaturation can be directly attributed to palmitoleic and not its elongation products, 18:1 cis-11 or 20:1 cis-13.

Keyword: lipogenesis

Alternation of plasma fatty acids composition and desaturase activities in children with liver steatosis.

The aim of this study was to investigate changes in plasma fatty acids proportions and estimated desaturase activities for variable grading of liver steatosis in children.In total, 111 schoolchildren (aged 8-18 years) were included in the analysis from March 2015 to August 2016. Anthropometric evaluation, liver ultrasound examination and scoring for nonalcoholic fatty liver disease (NAFLD score = 0-6), and biochemical and plasma fatty acids analysis were performed. We compared the composition ratio of fatty acids between children with high-grade liver steatosis (NAFLD score = 4-6), low-grade liver steatosis (NAFLD score = 1-3), and healthy controls (NAFLD score = 0). In addition, correlation coefficients (r) between NAFLD score, metabolic variables, and estimated activity of desaturase indices (stearoyl-coenzyme A desaturase-1 (SCD1), delta-5 and delta-6 desaturase) were calculated.Compared with healthy controls, children with liver steatosis showed a higher proportion of monounsaturated fatty acids (21.16 ± 2.81% vs. 19.68 ± 2.71%, p = 0.024). In addition, children with high- grade liver steatosis exhibited higher proportions of (C16:0), palmitoleic (C16:1n-7), dihomo-γ-linolenic (C20:3n-6), adrenic (C22:4n-6), and docosapentaenoic (C22:5n-6); and lower proportions of eicosapentaenoic (C20:5n-3) (P< 0.05). In all subjects, the NAFLD score was positively correlated with body mass index (BMI) (kg/m2) (r = 0.696), homeostasis model of assessment ratio-index (HOMA-IR) (r = 0.510), SCD1(16) (r = 0.273), and the delta-6 index (r = 0.494); and inversely associated with the delta-5 index (r = -0.443).Our current data suggested that children with liver steatosis was highly associated with obesity, and insulin resistance. In addition, increased endogenous through altered desaturase activity may contribute to the progression of liver steatosis in children.

Keyword: lipogenesis

Suppression of by valproic through repression of USF1-activated fatty synthesis in adipocytes.

VPA (valproic ), a short-chain fatty that is a HDAC (histone deacetylase) inhibitor, is known to suppress . In the present study, we identified the molecular mechanism of VPA-mediated suppression of in adipocytes. VPA suppressed the accumulation of intracellular triacylglycerol. The expression levels of PPARγ (peroxisome-proliferator-activated receptor γ) and C/EBPα (CCAAT/enhancer-binding protein α), which are key regulators of , as well as the expression of SCD (stearoyl-CoA desaturase), were decreased by the treatment with VPA. Moreover, glycerol release was decreased in the VPA-treated cells, even though the transcription levels of ATGL (adipose triacylglycerol lipase), HSL (hormone-sensitive lipase) and MGL (monoacylglycerol lipase), all of which are involved in lipolysis, were elevated by the treatment with VPA. It is noteworthy that the expression level of FAS (fatty synthase) was significantly suppressed when the cells were cultured in medium containing VPA. Furthermore, VPA-mediated suppression of the accumulation of the intracellular triacylglycerols was prevented by the treatment with , a major product of FAS. The results of promoter-luciferase and chromatin immunoprecipitation assays demonstrated that USF1(upstream stimulating factor 1) bound to the E-box of the promoter region of the FAS gene. In addition, the expression of USF1 was decreased by the treatment with VPA. siRNA-mediated knockdown of the expression of the USF1 gene repressed along with the decreased expression of the FAS gene. The overexpression of USF1 enhanced both and the expression of FAS in VPA-treated cells. These results indicate that VPA suppressed through the down-regulation of USF1-activated fatty synthesis in adipocytes.

Keyword: lipogenesis

Insulin-independent regulation of hepatic triglyceride synthesis by fatty acids.

A central paradox in type 2 diabetes is the apparent selective nature of hepatic insulin resistance--wherein insulin fails to suppress hepatic glucose production yet continues to stimulate , resulting in hyperglycemia, hyperlipidemia, and hepatic steatosis. Although efforts to explain this have focused on finding a branch point in insulin signaling where hepatic glucose and lipid metabolism diverge, we hypothesized that hepatic triglyceride synthesis could be driven by substrate, independent of changes in hepatic insulin signaling. We tested this hypothesis in rats by infusing [U-(13)C] palmitate to measure rates of fatty esterification into hepatic triglyceride while varying plasma fatty and insulin concentrations independently. These experiments were performed in normal rats, high fat-fed insulin-resistant rats, and insulin receptor 2\'-O-methoxyethyl chimeric antisense oligonucleotide-treated rats. Rates of fatty esterification into hepatic triglyceride were found to be dependent on plasma fatty infusion rates, independent of changes in plasma insulin concentrations and independent of hepatocellular insulin signaling. Taken together, these results obviate a paradox of selective insulin resistance, because the major source of hepatic lipid synthesis, esterification of preformed fatty acids, is primarily dependent on substrate delivery and largely independent of hepatic insulin action.

Keyword: lipogenesis

Prospective association of fatty acids in the de novo pathway with risk of type 2 diabetes: the Cardiovascular Health Study.

Experimental evidence suggests that hepatic de novo (DNL) affects insulin homeostasis via synthesis of saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs). Few prospective studies have used fatty biomarkers to assess associations with type 2 diabetes.We investigated associations of major circulating SFAs [ (16:0) and stearic (18:0)] and MUFA [oleic (18:1n-9)] in the DNL pathway with metabolic risk factors and incident diabetes in community-based older U.S. adults in the Cardiovascular Health Study. We secondarily assessed other DNL fatty biomarkers [myristic (14:0), palmitoleic (16:1n-7), 7-hexadecenoic (16:1n-9), and vaccenic (18:1n-7)] and estimated dietary SFAs and MUFAs.In 3004 participants free of diabetes, plasma phospholipid fatty acids were measured in 1992, and incident diabetes was identified by medication use and blood glucose. Usual diets were assessed by using repeated food-frequency questionnaires. Multivariable linear and Cox regression were used to assess associations with metabolic risk factors and incident diabetes, respectively.At baseline, circulating and stearic were positively associated with adiposity, triglycerides, inflammation biomarkers, and insulin resistance (P-trend < 0.01 each), whereas oleic showed generally beneficial associations (P-trend < 0.001 each). During 30,763 person-years, 297 incident diabetes cases occurred. With adjustment for demographics and lifestyle, (extreme-quintile HR: 1.89; 95% CI: 1.27, 2.83; P-trend = 0.001) and stearic (HR: 1.62; 95% CI: 1.09, 2.41; P-trend = 0.006) were associated with higher diabetes risk, whereas oleic was not significantly associated. In secondary analyses, vaccenic was inversely associated with diabetes (HR: 0.56; 95% CI: 0.38, 0.83; P-trend = 0.005). Other fatty biomarkers and estimated dietary SFAs or MUFAs were not significantly associated with incident diabetes.In this large prospective cohort, circulating and stearic were associated with higher diabetes risk, and vaccenic was associated with lower diabetes risk. These results indicate a need for additional investigation of biological mechanisms linking specific fatty acids in the DNL pathway to the pathogenesis of diabetes.ClinicalTrials.gov .© 2015 American Society for Nutrition.

Keyword: lipogenesis

Increased intramuscular lipid synthesis and low saturation relate to insulin sensitivity in endurance-trained athletes.

Intramuscular triglyceride (IMTG) has received considerable attention as a potential mechanism promoting insulin resistance. Endurance-trained athletes have high amounts of IMTG but are insulin sensitive, suggesting IMTG content alone does not change insulin action. Recent data suggest increased muscle lipid synthesis protects against fat-induced insulin resistance. We hypothesized that rates of IMTG synthesis at rest would be increased in athletes compared with controls. Eleven sedentary men and 11 endurance-trained male cyclists participated in this study. An intravenous glucose tolerance test was performed to assess insulin action. After 3 days of dietary control and an overnight fast, [13C16]palmitate was infused at 0.0174 micromol.kg(-1).min(-1) for 4 h, followed by a muscle biopsy to measure isotope incorporation into IMTG and diacylglycerol. Compared with controls, athletes were twice as insulin sensitive (P=0.004) and had a significantly greater resting IMTG concentration (athletes: 20.4+/-1.6 microg IMTG/mg dry wt, controls: 14.5+/-1.8 microg IMTG/mg dry wt, P=0.04) and IMTG fractional synthesis rate (athletes: 1.56+/-0.37%/h, controls: 0.61+/-0.15%/h, P=0.03). Stearoyl-CoA desaturase 1 mRNA expression (P=0.02) and protein content (P=0.03) were also significantly greater in athletes. Diacylglycerol, but not IMTG, saturation was significantly less in athletes compared with controls (P=0.002). These data indicate endurance-trained athletes have increased synthesis rates of skeletal muscle IMTG and decreased saturation of skeletal muscle diacylglycerol. Increased synthesis rates are not due to recovery from exercise and are likely adaptations to chronic endurance exercise training.

Keyword: lipogenesis

Macrophages with a deletion of the () gene have a more proinflammatory phenotype.

Phosphoenolpyruvate carboxykinase (Pck1) is a metabolic enzyme that is integral to the gluconeogenic and glyceroneogenic pathways. However, Pck1\'s role in macrophage metabolism and function is unknown. Using stable isotopomer MS analysis in a mouse model with a myeloid cell-specific deletion, we show here that this deletion increases the proinflammatory phenotype in macrophages. Incubation of LPS-stimulated bone marrow-derived macrophages (BMDM) with [U-C]glucose revealed reduced C labeling of citrate and malate and increased C labeling of lactate in Pck1-deleted bone marrow-derived macrophages. We also found that the Pck1 deletion in the myeloid cells increases reactive oxygen species (ROS). Of note, this altered macrophage metabolism increased expression of the M1 cytokines TNFα, IL-1β, and IL-6. We therefore conclude that contributes to M1 polarization in macrophages. Our findings provide important insights into the factors determining the macrophage inflammatory response and indicate that Pck1 activity contributes to metabolic reprogramming and polarization in macrophages.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: lipogenesis

Overexpression of apolipoprotein A-I alleviates endoplasmic reticulum stress in hepatocytes.

Abnormal lipid metabolism may contribute to an increase in endoplasmic reticulum (ER) stress, resulting in the pathogenesis of non-alcoholic steatohepatitis. Apolipoprotein A-I (apoA-I) accepts cellular free cholesterol and phospholipids transported by ATP-binding cassette transporter A1 to generate nascent high density lipoprotein particles. Previous studies have revealed that the overexpression of apoA-I alleviated hepatic lipid levels by modifying lipid transport. Here, we examined the effects of apoA-I overexpression on ER stress and genes involved in in both HepG2 cells and mouse hepatocytes.Human apoA-I was overexpressed in HepG2 hepatocytes, which were then treated with 2\xa0μg/mL tunicamycin or 500\xa0μM . Eight-week-old male apoA-I transgenic or C57BL/6 wild-type mice were intraperitoneally injected with 1\xa0mg/kg body weight tunicamycin or with saline. At 48\u2009h after injecting, blood and liver samples were collected.The overexpression of apoA-I in the models above resulted in decreased protein levels of ER stress makers and lipogenic gene products, including sterol regulatory element binding protein 1, fatty synthase, and acetyl-CoA carboxylase 1. In addition, the cellular levels of triglycerides and free cholesterol also decreased. Some of gene products which are related to ER stress-associated apoptosis were also affected by apoA-I overexpression. These results suggested that apoA-I overexpression could reduce steatosis by decreasing lipid levels and by suppressing ER stress and in hepatocytes.ApoA-I expression could significantly reduce hepatic ER stress and in hepatocytes.

Keyword: lipogenesis

Changes in lipid metabolism associated gene transcripts during porcine .

Pigs are recognised as suitable biomedical models to study obesity and obesity-related diseases; however, little is known about adipose tissue development and in pigs. In this study, the temporal expression of key genes involved in lipid metabolism was investigated during porcine and the metabolic fate of exogenously administered (16:0) was examined in differentiating preadipocytes. The expression of genes encoding elongases and desaturases increased simultaneously with those involved in fatty and triacylglycerol synthesis during porcine , and a high biosynthesis of monounsaturated fatty acids was measured prior to storage in differentiating preadipocytes. Although the total fatty oxidation in differentiating preadipocytes was low, differentiating cells showed increased expression of hormone-sensitive lipase and mitochondrial and peroxisomal genes. These data provide new insight into the temporal expression of genes involved in lipid metabolism during porcine and suggest a possible role of elongation and desaturation events prior to lipid accumulation in porcine adipocytes.

Keyword: lipogenesis

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile Metabolism and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) metabolism. However, whether TMAO aggravates liver steatosis by modulating BA metabolism and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA metabolism. Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: lipogenesis

Metabolic pathways promoting intrahepatic fatty accumulation in methionine and choline deficiency: implications for the pathogenesis of steatohepatitis.

The pathological mechanisms that distinguish simple steatosis from steatohepatitis (or NASH, with consequent risk of cirrhosis and hepatocellular cancer) remain incompletely defined. Whereas both a methionine- and choline-deficient diet (MCDD) and a choline-deficient diet (CDD) lead to hepatic triglyceride accumulation, MCDD alone is associated with hepatic insulin resistance and inflammation (steatohepatitis). We used metabolic tracer techniques, including stable isotope ([¹³C₄]palmitate) dilution and mass isotopomer distribution analysis (MIDA) of [¹³C₂]acetate, to define differences in intrahepatic fatty metabolism that could explain the contrasting effect of MCDD and CDD on NASH in C57Bl6 mice. Compared with control-supplemented (CS) diet, liver triglyceride pool sizes were similarly elevated in CDD and MCDD groups (24.37 ± 2.4, 45.94 ± 3.9, and 43.30 ± 3.5 μmol/liver for CS, CDD, and MCDD, respectively), but intrahepatic neutrophil infiltration and plasma alanine aminotransferase (31 ± 3, 48 ± 4, 231 ± 79 U/l, P < 0.05) were elevated only in MCDD mice. However, despite loss of peripheral fat in MCDD mice, neither the rate of appearance of palmitate (27.2 ± 3.5, 26.3 ± 2.3, and 28.3 ± 3.5 μmol·kg⁻¹·min⁻¹) nor the contribution of circulating fatty acids to the liver triglyceride pool differed between groups. Unlike CDD, MCDD had a defect in hepatic triglyceride export that was confirmed using intravenous tyloxapol (142 ± 21, 122 ± 15, and 80 ± 7 mg·kg⁻¹·h⁻¹, P < 0.05). Moreover, hepatic de novo was significantly elevated in the MCDD group only (1.4 ± 0.3, 2.3 ± 0.4, and 3.4 ± 0.4 μmol/day, P < 0.01). These findings suggest that important alterations in hepatic fatty metabolism may promote the development of steatohepatitis. Similar mechanisms may predispose to hepatocyte damage in human NASH.

Keyword: lipogenesis

A brominated flame retardant 2,2\',4,4\' tetrabrominated diphenyl ether (BDE-47) leads to in the copepod Tigriopus japonicus.

De novo (DNL) is a fatty synthesis process that requires several genes, including sterol regulatory element binding protein (SREBP), ATP-citrate lyase (ACLY), and acetyl-CoA carboxylase (ACC). DNL up-regulation is able to induce fat accumulation through an increase in fatty acids. To investigate the relationship between DNL up-regulation and the accumulation of fatty acids and lipid droplets in response to 2,2\',4,4\' tetrabrominated diphenyl ether (BDE-47), we examined DNL in the copepod Tigriopus japonicus. Transcription levels of DNL-related genes were increased after exposure to 2.5μg/L BDE-47 for 24h. After exposure to 2.5μg/L BDE-47, was significantly increased (P<0.05) at days 1 and 4, along with upregulation of fatty synthesis-related genes (e.g., desaturases and elongases). However, docosahexaenoic and arachidonic were down-regulated at days 1 and 4, showing an antagonistic effect. Lipid droplet area significantly increased in Nile red staining analysis after 24h of exposure to 2.5μg/L BDE-47 in T. japonicus, while DNL was down-regulated in response to 500μM salicylate (a inhibitor), indicating that BDE-47 exposure is closely associated with an increase in fatty acids in this copepod. This study provides a better understanding of the effects of BDE-47 on DNL in copepods.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

High oleic/stearic fatty- desaturation index in cord plasma from infants of mothers with gestational diabetes.

Enhanced fatty- desaturation by stearoyl-CoA desaturase enzyme-1 (SCD1) is associated with obesity. This study determined desaturation in the cord plasma of newborns of mothers with and without gestational diabetes (GDM).Newborns of mothers with GDM (n=21) and without (control, n=22) were recruited. Cord plasma fatty- desaturation indices (palmitoleic/, oleic/stearic ratios) were compared, and correlated with anthropometrics and biochemical measures. A subset of very low-density lipoprotein (VLDL) desaturation indices were determined to approximate the liver SCD1 activity.The total oleic/stearic index was higher in GDM, despite adjustment for cord glucose concentrations. Among GDM and controls, the oleic/stearic index correlated with cord glucose concentrations (rs=0.36, P=0.02). Both palmitoleic/ and oleic/stearic indices correlated with waist circumference (r=0.47, P=0.001; r=0.37, P=0.01). The VLDL oleic/stearic index was higher in GDM.The elevated total oleic/stearic index suggests increased in GDM newborns. Factors in addition to glucose supply may influence fetal SCD1 activity.

Keyword: lipogenesis

Soraphen, an inhibitor of the acetyl-CoA carboxylase system, improves peripheral insulin sensitivity in mice fed a high-fat diet.

Inhibition of the acetyl-CoA carboxylase (ACC) system, consisting of the isozymes ACC1 and ACC2, may be beneficial for treatment of insulin resistance and/or obesity by interfering with de novo and beta-oxidation. We have evaluated effects of pharmacological inhibition of ACC by soraphen (SP) on high fat (HF) diet-induced insulin resistance in mice.Male C57Bl6/J mice were fed control chow, a HF diet or a HF diet supplemented with SP (50 or 100 mg/kg/day).Body weight gain and total body fat content of SP-treated animals were significantly reduced compared with HF-fed mice. Fractional synthesis of palmitate was significantly reduced in mice treated with SP, indicative for ACC1 inhibition. Plasma beta-hydroxybutyrate levels were significantly elevated by SP, reflecting simultaneous inhibition of ACC2 activity. Mice treated with SP showed improved peripheral insulin sensitivity, as assessed by hyperinsulinaemic euglycaemic clamps.Pharmacological inhibition of the ACC system is of potential use for treatment of key components of the metabolic syndrome.

Keyword: lipogenesis

Natural Abundance Carbon Isotopic Analysis Indicates the Equal Contribution of Local Synthesis and Plasma Uptake to Palmitate Levels in the Mouse Brain.

Saturated fatty acids are the most abundant fatty acids in the brain, however, there has been some debate regarding the ability of intact dietary saturated fatty acids to be incorporated into the brain. In the present study, we use compound specific isotope analysis to measure the natural abundance carbon isotopic signature of brain, liver, and blood (PAM) and compare it to the dietary PAM and sugar isotopic signatures to calculate the relative contribution of both the incorporation of intact and endogenously synthesized PAM to these pools. Mice were equilibrated to the study diet, and extracted fatty acids were analyzed with gas chromatography isotope ratio mass spectrometry to determine the carbon isotopic signature of PAM (δ C ). Liver, serum total, and serum unesterified fatty δ C ranged between -20.6 and -21.1 mUr and were approximately 8.5 mUr more enriched in C when compared to the dietary PAM signature. Brain δ C was found to be more enriched than liver or blood pools (-16.7 ± 0.2 mUr, mean ± SD). Two end-member-mixed modeling using the carbon isotopic signature of dietary PAM and dietary sugars determined the contribution of synthesis to the total tissue PAM pool to range between 44% and 48%. This suggests that endogenous synthesis and dietary PAM are near equal contributors to brain, liver, and blood PAM pools. In conclusion, our data provide evidence that brain PAM levels are maintained by both local endogenous synthesis and through the uptake of intact PAM from the blood.© 2018 AOCS.

Keyword: lipogenesis

Inhibitory effects of high stability fucoxanthin on -induced lipid accumulation in human adipose-derived stem cells through modulation of long non-coding RNA.

Obesity is a serious worldwide disease, which is growing in epidemic proportions. Adipose-derived stem cells (ADSCs) are characterized as a source of mesenchymal stem cells that have acted as a potential application for regeneration. Recently, seaweeds rich in flavonoids and polysaccharides have been supposed to show the ability to modulate risk factors for obesity and related diseases. In the present study, we investigated the anti-obesity properties of high stability fucoxanthin (HS-Fx) derived from brown seaweeds on the of ADSCs upon treatment with (PA). First, we showed the differentiation capability of ADSCs from morbid obesity patients to transform into different cell types. Second, we found that the co-treatment of ADSCs with HS-Fx and PA showed no significant cytotoxicity against ADSCs, but PA induced the elevation of reactive oxygen species (ROS) and lipid droplet accumulation was abolished. Thirdly, the PA-mediated down-regulation of lipid metabolism genes was reversed by the treatment of HS-Fx. By long non-coding RNAs (lncRNAs) screening, we found that PA-induced increases in the targeted lncRNAs were also decreased upon treatment with HS-Fx. On Silencing, these lncRNAs corresponded to the decrease in the lipid droplet accumulation of ADSCs induced by PA. ADSCs from obese patients would be direct and meaningful model cells to investigate the development of obesity-related diseases and their treatments, rather than cell lines from other species. HS-Fx showed anti-obesity capability through modulating the elevation of ROS, down-regulation of lipid metabolism genes induced by PA, and upstream signaling, which might be critically resulted from the expression of lncRNAs.

Keyword: lipogenesis

Effect of linseed addition on the expression of some lipid metabolism genes in the adipose tissue of young Italian Simmental and Holstein bulls.

The objective of this trial was to determine the effect of breed and long-term dietary linseed addition on composition of fatty acids and expression of some genes involved in the lipid metabolism within subcutaneous (s.c.) adipose tissue of young bulls. Italian Simmental and Holstein bulls (n=16/breed) were fed a corn silage-grass hay diet with or without 8% (DM basis) whole ground linseed. Inclusion of linseed, rich in α-linoleic (C18:3n-3), increased (P<0.05) the proportions of linolelaidic (C18:2n-6trans), γ-linolenic (C18:3n-6), C18:3n-3, and rumenic (cis9,trans11 conjugated linoleic ) acids, as well as total n-3 fatty , total PUFA, and PUFA:SFA, but decreased (P<0.05) weight percentages of myristic (C14:0), pentadecanoic (C15:0), (C16:0), palmitelaidic (C16:1n-9trans), and margaric (C17:0) acids, along with n-6:n-3, in the s.c. fat of young bulls. Even though PUFA were similar (P≥0.23) between bull breeds, s.c. fat from Holstein bulls had greater (P<0.05) proportions of tridecylic (C13:0), myristoleic (C14:1) and palmitoleic (C16:1n-9cis) acids and a lower (P<0.05) proportion of margaric (C17:0) than s.c. fat from Simmental bulls. Feeding linseed decreased (P<0.05) the expression of stearoyl CoA desaturase (SCD) and the lipoprotein lipase (LPL) gene without affecting (P≥0.19) fatty synthase (FASN), leptin (LEP), and PPARγ2 mRNA in the s.c. fat of bulls; however, there was no effect of bull breed (P≥0.11) or interactive effect of breed and linseed (P≥0.23) on gene expression. Expression of PPARγ2 was positively correlated with SCD (r=0.454; P=0.01), LEP (r=0.500; P<0.01), and LPL (r=0.531; P<0.01) mRNA, indicating that PPARγ2 increases the expression of genes involved in .

Keyword: lipogenesis

-induced apoptosis in pancreatic β-cells is increased by liver X receptor agonist and attenuated by eicosapentaenoate.

Saturated fatty acids are implicated in the development of diabetes via the impairment of pancreatic islet β-cell viability and function. Liver X receptors (LXRs) and eicosapentaenoate (EPA) are known regulators of fatty metabolism. However, their roles in the pathogenesis of diabetes remain incompletely understood. The aim of this study was to determine the effects of EPA and the LXR agonist T0901317 on saturated fatty ()-induced apoptosis in the insulinoma β-cell line INS-1, a model for insulin-secreting β-cells. T0901317 significantly promoted -induced apoptotic cell death in the INS-1 cells. Consistent with these results, caspase-3 activity and BAX and sterol regulatory element binding protein-1c (SREBP-1c) mRNA levels were markedly increased in INS-1 cells co-administered and T0901317. The production of reactive oxygen species was considerably higher in the cells cultured concurrently with T0901317 and than in the cells incubated with either agent alone. EPA treatment attenuated the cellular death promoted by and T0901317 in the INS-1 cells, disclosing a possible mediating mechanism involving the inhibition of SREBP-1c. Finally, T0901317 up-regulated the -induced expression of p27(KIP1), transforming growth factor beta 1, and SMAD3 proteins in INS-1 cells. These results demonstrate that -induced apoptosis in β-cells is enhanced by T0901317 via the activation of LXRs and is blocked by EPA via the inhibition of SREBP-1c, suggesting that the regulation of and lipotoxicity affecting pancreatic β-cell viability and insulin production may be a unique strategy for diabetes therapy.

Keyword: lipogenesis

Procyanidin B2 ameliorates free fatty acids-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state.

Procyanidin B2, a naturally occurring phenolic compound, has been reported to exert multiple beneficial functions. However, the effect of procyanidin B2 on free fatty acids (FFAs)-induced hepatic steatosis remains obscure. The present study is therefore aimed to elucidate the protective effect of procyanidin B2 against hepatic steatosis and its underlying mechanism. Herein, we reported that procyanidin B2 attenuated FFAs-induced lipid accumulation and its associated oxidative stress by scavenging excessive ROS and superoxide anion radicals, blocking loss of mitochondrial membrane potential, restoring glutathione content, and increasing activity of antioxidant enzymes (GPx, SOD and CAT) in hepatocytes. Procyanidin B2 mechanistically promoted lipid degradation via modulation of transcription factor EB (TFEB), a master regulator of lysosomal pathway. Molecular docking analysis indicated a possible ligand-binding position of procyanidin B2 with TFEB. In addition, administration of procyanidin B2 resulted in a significant reduction of hepatic fat accumulation in high-fat diet (HFD)-induced obese mice, and also ameliorated HFD-induced metabolic abnormalities, including hyperlipidemia and hyperglycemia. It was confirmed that procyanidin B2 prevented HFD-induced hepatic fat accumulation through down-regulating -related gene expressions (PPARγ, C/EBPα and SREBP-1c), inhibiting pro-inflammatory cytokines production (IL-6 and TNF-α) and increasing antioxidant enzymes activity (GPx, SOD and CAT). Moreover, hepatic fatty acids analysis indicated that procyanidin B2 caused a significant increase in the levels of , oleic and linoleic . Intriguingly, procyanidin B2 restored the decreased nuclear TFEB expression in HFD-induced liver steatosis and up-regulated its target genes involved in lysosomal pathway (Lamp1, Mcoln, Uvrag), which suggested a previously unrecognized mechanism of procyanidin B2 on ameliorating HFD-induced hepatic steatosis. Taken together, our results demonstrated that procyanidin B2 attenuated FFAs-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state, which had important implications that modulation of TFEB might be a potential therapeutic strategy for hepatic steatosis and procyanidin B2 could represent a promising novel agent in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD).Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Osteonecrosis of the Femoral Head: Lipotoxicity Exacerbation in MSC and Modifications of the Bone Marrow Fluid.

Osteonecrosis of the femoral head (ON) is a multifactorial bone disease that can evolve to a progressive destruction of the hip joint. Different pathogenic processes have been proposed, among them, an increase of bone marrow (BM) fat resulting from adipocyte accumulation. Marrow adipocytes are active BM residents that influence the microenvironment by releasing cytokines, adipokines, and free fatty acids (FA). We explored the impact of palmitate (Palm) and oleate on function and survival of BM-derived mesenchymal stromal cells (MSC) of osteonecrotic patients (ONMSC) and healthy volunteers. Moreover, we analyzed the FA profile of the serum and the BM supernatant fluid (BMSF). We demonstrated that exposure to the saturated FA Palm favored MSC differentiation through the adipogenic lineage at the expense of the osteoblastic phenotype. Moreover, was intensified in ONMSC. The susceptibility to Palm toxicity was aggravated in ONMSC concomitantly with a greater activation of the proapoptotic extracellular signal-regulated kinase pathway. Moreover, cellular mechanisms implicated in the protection against lipotoxicity, such as stearoyl-coenzyme A desaturase 1 and carnitine palmitoyl transferase 1 expression, were dysregulated in ONMSC. Palm-induced interleukin (IL)-6 and IL-8 secretion was also exacerbated in ONMSC. Our results established that, in the serum, the FA profiles were comparable in ON and healthy subjects. However, both the concentrations and the FA composition were modified in the BMSF of ON patients, highlighting a drastic change of the BM microenvironment in ON patients. Altogether, our work suggests that marrow adipocyte enlargement could affect the process of bone remodeling and, therefore, play a role in the pathogenesis of ON.Copyright © 2017 by the Endocrine Society.

Keyword: lipogenesis

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting and inflammation.

Non-alcoholic fatty liver disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited lipid accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), fatty synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased lipid accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: lipogenesis

Polymorphism in the ELOVL6 gene is associated with a major QTL effect on fatty composition in pigs.

The ELOVL fatty elongase 6 (ELOVL6), the only elongase related to de novo , catalyzes the rate-limiting step in the elongation cycle by controlling the fatty balance in mammals. It is located on pig chromosome 8 (SSC8) in a region where a QTL affecting , and palmitoleic composition was previously detected, using an Iberian x Landrace intercross. The main goal of this work was to fine-map the QTL and to evaluate the ELOVL6 gene as a positional candidate gene affecting the percentages of and palmitoleic fatty acids in pigs.The combination of a haplotype-based approach and single-marker analysis allowed us to identify the main, associated interval for the QTL, in which the ELOVL6 gene was identified and selected as a positional candidate gene. A polymorphism in the promoter region of ELOVL6, ELOVL6:c.-533C>T, was highly associated with the percentage of and palmitoleic acids in muscle and backfat. Significant differences in ELOVL6 gene expression were observed in backfat when animals were classified by the ELOVL6:c.-533C>T genotype. Accordingly, animals carrying the allele associated with a decrease in ELOVL6 gene expression presented an increase in C16:0 and C16:1(n-7) fatty content and a decrease of elongation activity ratios in muscle and backfat. Furthermore, a SNP genome-wide association study with ELOVL6 relative expression levels in backfat showed the strongest effect on the SSC8 region in which the ELOVL6 gene is located. Finally, different potential genomic regions associated with ELOVL6 gene expression were also identified by GWAS in liver and muscle, suggesting a differential tissue regulation of the ELOVL6 gene.Our results suggest ELOVL6 as a potential causal gene for the QTL analyzed and, subsequently, for controlling the overall balance of fatty composition in pigs.

Keyword: lipogenesis

Dietary carnosic suppresses hepatic steatosis formation via regulation of hepatic fatty metabolism in high-fat diet-fed mice.

In this study, we examined the hepatic anti-steatosis activity of carnosic (CA), a phenolic compound of rosemary (Rosmarinus officinalis) leaves, as well as its possible mechanism of action, in a high-fat diet (HFD)-fed mice model. Mice were fed a HFD, or a HFD supplemented with 0.01% (w/w) CA or 0.02% (w/w) CA, for a period of 12 weeks, after which changes in body weight, blood lipid profiles, and fatty mechanism markers were evaluated. The 0.02% (w/w) CA diet resulted in a marked decline in steatosis grade, as well as in homeostasis model assessment of insulin resistance (HOMA-IR) index values, intraperitoneal glucose tolerance test (IGTT) results, body weight gain, liver weight, and blood lipid levels (P < 0.05). The expression level of hepatic lipogenic genes, such as sterol regulating element binding protein-1c (SREBP-1c), liver-fatty binding protein (L-FABP), stearoyl-CoA desaturase 1 (SCD1), and fatty synthase (FAS), was significantly lower in mice fed 0.01% (w/w) CA and 0.02% (w/w) CA diets than that in the HFD group; on the other hand, the expression level of β-oxidation-related genes, such as peroxisome proliferator-activated receptor α (PPAR-α), carnitine palmitoyltransferase 1 (CPT-1), and acyl-CoA oxidase (ACO), was higher in mice fed a 0.02% (w/w) CA diet, than that in the HFD group (P < 0.05). In addition, the hepatic content of (C16:0), palmitoleic (C16:1), and oleic (C18:1) was significantly lower in mice fed the 0.02% (w/w) CA diet than that in the HFD group (P < 0.05). These results suggest that orally administered CA suppressed HFD-induced hepatic steatosis and fatty liver-related metabolic disorders through decrease of de novo and fatty elongation and increase of fatty β-oxidation in mice.

Keyword: lipogenesis

Fatty synthesis is a therapeutic target in human liposarcoma.

Liposarcomas (LS) are mesenchymal tumors that can recur after surgical resection and often do not respond to presently available medical therapies. This study demonstrates the dependence of LS on de novo long-chain fatty synthesis for growth. can be impaired by inhibiting the activities of lipogenic enzymes, including acetyl CoA-carboxylase (ACC) and fatty synthase (FASN), or by suppressing the expression of key genes involved in the pathway and its regulation. The FASN inhibitors cerulenin and orlistat reduced the growth of two LS cell lines (LiSa2, SW872), as did inhibition of ACC with soraphen A. CDDO-Me, a synthetic triterpenoid, suppressed expression of Spot 14 and FASN genes and likewise inhibited LS cell growth. Importantly, the anti-proliferative effect of each agent was prevented by the co-administration of palmitate, the major product of cellular long-chain fatty synthesis. In stark contrast to LS cells, these compounds had no effect on the growth of fibroblasts. Four biochemically distinct agents that target critical points in the fatty synthetic pathway exert anti-proliferative effects on LS cells, and rescue of cell growth by suggests that reduced tumor cell mediates the growth inhibition. These findings warrant further studies aimed at the clinical exploitation of the dependence of LS cell growth on fatty acids.

Keyword: lipogenesis

Using [H]water to quantify the contribution of de novo palmitate synthesis in plasma: enabling back-to-back studies.

An increased contribution of de novo (DNL) may play a role in cases of dyslipidemia and adipose accretion; this suggests that inhibition of fatty synthesis may affect clinical phenotypes. Since it is not clear whether modulation of one step in the lipogenic pathway is more important than another, the use of tracer methods can provide a deeper level of insight regarding the control of metabolic activity. Although [H]water is generally considered a reliable tracer for quantifying DNL in vivo (it yields a homogenous and quantifiable precursor labeling), the relatively long half-life of body water is thought to limit the ability of performing repeat studies in the same subjects; this can create a bottleneck in the development and evaluation of novel therapeutics for inhibiting DNL. Herein, we demonstrate the ability to perform back-to-back studies of DNL using [H]water. However, this work uncovered special circumstances that affect the data interpretation, i.e., it is possible to obtain seemingly negative values for DNL. Using a rodent model, we have identified a physiological mechanism that explains the data. We show that one can use [H]water to test inhibitors of DNL by performing back-to-back studies in higher species [i.e., treat nonhuman primates with platensimycin, an inhibitor of fatty synthase]; studies also demonstrate the unsuitability of [C]acetate.

Keyword: lipogenesis

Selective cannabinoid-1 receptor blockade benefits fatty and triglyceride metabolism significantly in weight-stable nonhuman primates.

The goal of this study was to determine whether administration of the CB₁ cannabinoid receptor antagonist rimonabant would alter fatty flux in nonhuman primates. Five adult baboons (Papio Sp) aged 12.1 ± 4.7 yr (body weight: 31.9 ± 2.1 kg) underwent repeated metabolic tests to determine fatty and TG flux before and after 7 wk of treatment with rimonabant (15 mg/day). Animals were fed ad libitum diets, and stable isotopes were administered via diet (d₃₁-tripalmitin) and intravenously (¹³C₄-palmitate, ¹³C₁-acetate). Plasma was collected in the fed and fasted states, and blood lipids were analyzed by GC-MS. DEXA was used to assess body composition and a hyperinsulinemic euglycemic clamp used to assess insulin-mediated glucose disposal. During the study, no changes were observed in food intake, body weight, plasma, and tissue endocannabinoid concentrations or the quantity of liver-TG fatty acids originating from de novo (19 ± 6 vs. 16 ± 5%, for pre- and posttreatment, respectively, P = 0.39). However, waist circumference was significantly reduced 4% in the treated animals (P < 0.04), glucose disposal increased 30% (P = 0.03), and FFA turnover increased 37% (P = 0.02). The faster FFA flux was consistent with a 43% reduction in these fatty acids used for TRL-TG synthesis (40 ± 3 vs. 23 ± 4%, P = 0.02) and a twofold increase in TRL-TG turnover (1.5 ± 0.9 vs. 3.1 ± 1.4 μmol·kg⁻¹·h⁻¹, P = 0.03). These data support the potential for a strong effect of CB₁ receptor antagonism at the level of adipose tissue, resulting in improvements in fasting turnover of fatty acids at the whole body level, central adipose storage, and significant improvements in glucose homeostasis.

Keyword: lipogenesis

Energy and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of energy from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body energy and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net energy and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible energy basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty constituents are , oleic, and linoleic ), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine energy and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross energy intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated energy retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk fat yield and milk energy output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable energy intake, increased milk fat yield and milk energy output, increased plasma triacylglycerides and long-chain fatty concentrations, and had no effect on energy retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable energy intake, and total milk, fat, protein, and lactose yields. Milk energy output increased at the HMP level, and protein level did not affect total energy retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted energy retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra energy intake into milk and had no effect on milk N efficiency.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Fructose and glucose combined with free fatty acids induce metabolic disorders in HepG2 cell: A new model to study the impacts of high-fructose/sucrose and high-fat diets in vitro.

This work investigated the underlying mechanism of high-fructose/sucrose and high-fat diets, which rapidly induce metabolic syndrome in vivo, via a new cell model.Glucose and/or fructose were used to induce the human hepatoma cell (HepG2) in the presence of , oleic , or combined fatty acids (CFA) for 24 h. The alterations in lipid and uric production, glucose metabolism, oxidative status, and related genes and proteins were monitored. The cell model that featured metabolic disorders was established by treatment of 10 mM glucose and 15 mM fructose plus 1 mM CFA. Results showed that mainly induced insulin resistance, oxidative stress, and triglyceride (TG) secretion, whereas oleic mainly contributed to intracellular TG. Fructose was mainly responsible for uric and cholesterol production. In addition, fructose synergistically elevated the intra- and extracellular TG and extracellular malonaldehyde with glucose and CFA. Regulations of genes and proteins associated with carbohydrate metabolism and partially explained the action of fructose in inducing the metabolic disorders in cell.The combination of glucose, fructose, and CFA could successfully induce metabolic disorders in HepG2 cells, including dyslipidemia, insulin resistance, hyperuricemia, and oxidative stress.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: lipogenesis

Optimization of pineapple pulp residue hydrolysis for lipid production by Rhodotorula glutinis TISTR5159 using as biodiesel feedstock.

The higher lipid productivity of Rhodotorula glutinis TISTR5159 was achieved by optimizing the pineapple pulp hydrolysis for releasing the high sugars content. The sequential simplex method operated by varied; solid-to-liquid ratio, sulfuric concentration, temperature, and hydrolysis time were successfully applied and the highest sugar content (83.2\xa0g/L) evaluated at a solid-to-liquid ratio of 1:10.8, 3.2% sulfuric , 105\xa0°C for 13.9\xa0min. Moreover, the (NH4)2SO4 supplement enhanced the lipid productivity and gave the maximum yields of biomass and lipid of 15.2\xa0g/L and 9.15\xa0g/L (60.2%), respectively. The C16 and C18 fatty acids were found as main components included oleic (55.8%), (16.6%), linoleic (11.9%), and stearic (7.8%). These results present the possibility to convert the sugars in pineapple pulp hydrolysate to lipids. The fatty profile was also similar to vegetable oils. Thus, it could be used as potential feedstock for biodiesel production.

Keyword: lipogenesis

Toyocamycin attenuates free fatty -induced hepatic steatosis and apoptosis in cultured hepatocytes and ameliorates nonalcoholic fatty liver disease in mice.

A high serum level of saturated free fatty acids (FFAs) is associated with the development of nonalcoholic fatty liver disease (NAFLD). X-box binding protein-1 (XBP-1) is activated by FFA treatment upon splicing. XBP-1 is a transcription factor induced by the endoplasmic reticulum (ER) stress sensor endoribonuclease inositol-requiring enzyme 1 alpha (IRE1α). However, the role of XBP-1 in NAFLD remains relatively unexplored. Toyocamycin was recently reported to attenuate the activation of XBP-1, possibly by inducing a conformational change in IRE1α. In this study, we examined the effect of toyocamycin on hepatocyte lipoapoptosis and steatosis. We also explored the effects of toyocamycin in a mouse model of NAFLD.Huh-7 cells and isolated rat primary hepatocytes were treated with (PA), which is a saturated FFA, in the presence or absence of toyocamycin. In addition, male C57BL/6J mice were fed a diet rich in saturated fat, fructose, and cholesterol (FFC) for 4 months, after which the effect of toyocamycin was assessed.Toyocamycin attenuated FFA-induced steatosis. It also significantly reduced PA-induced hepatocyte lipoapoptosis. In addition, toyocamycin reduced the expression of cytosine-cytosine-adenosine-adenosine-thymidine enhancer-binding protein homologous protein (CHOP), which is a key player in ER stress-mediated apoptosis, as well as its downstream cell death modulator, death receptor 5. In the in vivo study, toyocamycin ameliorated the liver injury caused by FFC-induced NAFLD. It also reduced hepatic steatosis and the expression of lipogenic genes.The data we obtained suggest that toyocamycin attenuates hepatocyte and ameliorates NAFLD in vivo and may therefore be beneficial in the treatment of NAFLD in humans.

Keyword: lipogenesis

(16:0) competes with omega-6 linoleic and omega-3 ɑ-linolenic acids for FADS2 mediated Δ6-desaturation.

Sapienic , 16:1n-10 is the most abundant unsaturated fatty on human skin where its synthesis is mediated by FADS2 in the sebaceous glands. The FADS2 product introduces a double bond at the Δ6, Δ4 and Δ8 positions by acting on at least ten substrates, including 16:0, 18:2n-6, and 18:3n-3. Our aim was to characterize the competition for accessing FADS2 mediated Δ6 desaturation between 16:0 and the most abundant polyunsaturated fatty acids (PUFA) in the human diet, 18:2n-6 and 18:3n-3, to evaluate whether competition may be relevant in other tissues and thus linked to metabolic abnormalities associated with FADS2 or fatty levels. MCF7 cells stably transformed with FADS2 biosynthesize 16:1n-10 from exogenous 16:0 in preference to 16:1n-7, the immediate product of SCD highly expressed in cancer cell lines, and 16:1n-9 via partial β-oxidation of 18:1n-9. Increasing availability of 18:2n-6 or 18:3n-3 resulted in decreased bioconversion of 16:0 to 16:1n-10, simultaneously increasing the levels of highly unsaturated products. FADS2 cells accumulate the desaturation-elongation products 20:3n-6 and 20:4n-3 in preference to the immediate desaturation products 18:3n-6 and 18:4n-3 implying prompt/coupled elongation of the nascent desaturation products. MCF7 cells incorporate newly synthesized 16:1n-10 into phospholipids. These data suggest that excess 16:0 due to, for instance, de novo from high carbohydrate or alcohol consumption, inhibits synthesis of highly unsaturated fatty acids, and may in part explain why supplemental preformed EPA and DHA in some studies improves insulin resistance and other factors related to diabetes and metabolic syndrome aggravated by excess calorie consumption.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Fatty composition, fat deposition, lipogenic gene expression and performance of broiler fed diet supplemented with different sources of oil.

The present study assessed the effect of feeding palm oil (PO), sunflower oil (SO) and their combination on performance, fat deposition, fatty composition and lipogenic gene expression of broilers reared for 42\xa0days. A total of 144 1-day-old broilers (Cobb500) were randomly allotted into four treatment diets with each having six replicates of six chicks in each replicate following a completely randomized design. Live weight gain and feed efficiency was significantly (P\xa0<\xa00.05) higher in birds fed with a combination of oil sources compared to controls. Birds fed with the combination of oil and SO alone had higher carcass yield and lower abdominal fat. Higher (P\xa0<\xa00.05) concentrations of unsaturated fatty acids (UFA) and lower concentrations of and saturated fatty (SFA) was found in birds fed SO alone and combinations of SO and PO. Furthermore, the outcomes showed that birds fed diet supplemented with SO and the combination of SO and PO down-regulated gene expression of key hepatic lipogenic enzymes of fatty acids synthase (FAS), acetyl-CoA carboxylase (ACC) and stearoyl-CoA desaturase (SCD). These findings suggest that the diet containing the combination of 2% PO and 4% SO may reduce hepatic , as well as lower abdominal fat content of broilers.© 2017 Japanese Society of Animal Science.

Keyword: lipogenesis

New role of irisin in hepatocytes: The protective effect of hepatic steatosis in vitro.

Irisin is a newly identified myokine related to exercise and the browning of white fat. Recently, it was reported that irisin serum levels are associated with intrahepatic triglyceride content, suggesting that it might have an important role in the liver. The aim of this study was to determine the role of irisin in hepatocytes. Specifically, the effect of recombinant irisin on (PA)-induced and its related signal pathways were examined in AML12 cells and mouse primary hepatocytes. In the present study, we observed the presence of irisin inside the cells in response to the treatment of recombinant irisin by flow cytometry and cell imaging technique. Recombinant irisin significantly inhibited the PA-induced increase in lipogenic markers ACC and FAS at the mRNA and protein levels, and prevented the PA-induced lipid accumulation in hepatocytes. Additionally, irisin inhibited the PA-induced increase in the expression, nuclear localization, and transcriptional activities of the master regulators of (LXRα and SREBP-1c). Moreover, irisin attenuated PA-induced oxidative stress, which was confirmed by measuring the expression of inflammatory markers (NFκB, COX-2, p38 MAPK, TNF, IL-6) and superoxide indicator (dihydroethidium). The preventive effects of irisin against and oxidative stress were mediated by the inhibition of protein arginine methyltransferase-3 (PRMT3). These findings suggested that irisin might have a beneficial role in the prevention of hepatic steatosis by altering the expression of lipogenic genes and attenuating oxidative stress in a PRMT3 dependent manner.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Exercise performed immediately after fructose ingestion enhances fructose oxidation and suppresses fructose storage.

Exercise prevents the adverse effects of a high-fructose diet through mechanisms that remain unknown.We assessed the hypothesis that exercise prevents fructose-induced increases in very-low-density lipoprotein (VLDL) triglycerides by decreasing the fructose conversion into glucose and VLDL-triglyceride and fructose carbon storage into hepatic glycogen and lipids.Eight healthy men were studied on 3 occasions after 4 d consuming a weight-maintenance, high-fructose diet. On the fifth day, the men ingested an oral (13)C-labeled fructose load (0.75 g/kg), and their total fructose oxidation ((13)CO2 production), fructose storage (fructose ingestion minus (13)C-fructose oxidation), fructose conversion into blood (13)C glucose (gluconeogenesis from fructose), blood VLDL-(13)C palmitate (a marker of hepatic de novo ), and lactate concentrations were monitored over 7 postprandial h. On one occasion, participants remained lying down throughout the experiment [fructose treatment alone with no exercise condition (NoEx)], and on the other 2 occasions, they performed a 60-min exercise either 75 min before fructose ingestion [exercise, then fructose condition (ExFru)] or 90 min after fructose ingestion [fructose, then exercise condition (FruEx)].Fructose oxidation was significantly (P < 0.001) higher in the FruEx (80% ± 3% of ingested fructose) than in the ExFru (46% ± 1%) and NoEx (49% ± 1%). Consequently, fructose storage was lower in the FruEx than in the other 2 conditions (P < 0.001). Fructose conversion into blood (13)C glucose, VLDL-(13)C palmitate, and postprandial plasma lactate concentrations was not significantly different between conditions.Compared with sedentary conditions, exercise performed immediately after fructose ingestion increases fructose oxidation and decreases fructose storage. In contrast, exercise performed before fructose ingestion does not significantly alter fructose oxidation and storage. In both conditions, exercise did not abolish fructose conversion into glucose or its incorporation into VLDL triglycerides. This trial was registered at clinicaltrials.gov as .© 2016 American Society for Nutrition.

Keyword: lipogenesis

Loss of biliverdin reductase-A promotes lipid accumulation and lipotoxicity in mouse proximal tubule cells.

Obesity and increased lipid availability have been implicated in the development and progression of chronic kidney disease. One of the major sites of renal lipid accumulation is in the proximal tubule cells of the kidney, suggesting that these cells may be susceptible to lipotoxicity. We previously demonstrated that loss of hepatic biliverdin reductase A (BVRA) causes fat accumulation in livers of mice on a high-fat diet. To determine the role of BVRA in mouse proximal tubule cells, we generated a CRISPR targeting BVRA for a knockout in mouse proximal tubule cells (BVRA KO). The BVRA KO cells had significantly less metabolic potential and mitochondrial respiration, which was exacerbated by treatment with , a saturated fatty . The BVRA KO cells also showed increased intracellular triglycerides which were associated with higher fatty uptake gene cluster of differentiation 36 as well as increased de novo as measured by higher neutral lipids. Additionally, neutrophil gelatinase-associated lipocalin 1 expression, annexin-V FITC staining, and lactate dehydrogenase assays all demonstrated that BVRA KO cells are more sensitive to -induced lipotoxicity than wild-type cells. Phosphorylation of BAD which plays a role in cell survival pathways, was significantly reduced in -treated BVRA KO cells. These data demonstrate the protective role of BVRA in proximal tubule cells against saturated fatty -induced lipotoxicity and suggest that activating BVRA could provide a benefit in protecting from obesity-induced kidney injury.

Keyword: lipogenesis

PKA/AMPK signaling in relation to adiponectin\'s antiproliferative effect on multiple myeloma cells.

Obesity increases the risk of developing multiple myeloma (MM). Adiponectin is a cytokine produced by adipocytes, but paradoxically decreased in obesity, that has been implicated in MM progression. Herein, we evaluated how prolonged exposure to adiponectin affected the survival of MM cells as well as putative signaling mechanisms. Adiponectin activates protein kinase A (PKA), which leads to decreased AKT activity and increased AMP-activated protein kinase (AMPK) activation. AMPK, in turn, induces cell cycle arrest and apoptosis. Adiponectin-induced apoptosis may be mediated, at least in part, by the PKA/AMPK-dependent decline in the expression of the enzyme acetyl-CoA-carboxylase (ACC), which is essential to . Supplementation with , the preliminary end product of fatty synthesis, rescues MM cells from adiponectin-induced apoptosis. Furthermore, 5-(tetradecyloxy)-2-furancarboxylic (TOFA), an ACC inhibitor, exhibited potent antiproliferative effects on MM cells that could also be inhibited by fatty supplementation. Thus, adiponectin\'s ability to reduce survival of MM cells appears to be mediated through its ability to suppress . Our findings suggest that PKA/AMPK pathway activators, or inhibitors of ACC, may be useful adjuvants to treat MM. Moreover, the antimyeloma effect of adiponectin supports the concept that hypoadiponectinemia, as occurs in obesity, promotes MM tumor progression.

Keyword: lipogenesis

Exendin-4 improves steatohepatitis by increasing Sirt1 expression in high-fat diet-induced obese C57BL/6J mice.

The effects of exendin-4 on Sirt1 expression as a mechanism of reducing fatty liver have not been previously reported. Therefore, we investigated whether the beneficial effects of exendin-4 treatment on fatty liver are mediated via Sirt1 in high-fat (HF) diet-induced obese C57BL/6J mice and related cell culture models. Exendin-4 treatment decreased body weight, serum free fatty (FA), and triglyceride levels in HF-induced obese C57BL/6J mice. Histological analysis showed that exendin-4 reversed HF-induced hepatic accumulation of lipids and inflammation. Exendin-4 treatment increased mRNA and protein expression of Sirt1 and its downstream factor, AMPK, in vivo and also induced genes associated with FA oxidation and glucose metabolism. In addition, a significant increase in the hepatic expression of Lkb1 and Nampt mRNA was observed in exendin-4-treated groups. We also observed increased expression of phospho-Foxo1 and GLUT2, which are involved in hepatic glucose metabolism. In HepG2 and Huh7 cells, mRNA and protein expressions of GLP-1R were increased by exendin-4 treatment in a dose-dependent manner. Exendin-4 enhanced protein expression of Sirt1 and phospho-AMPKα in HepG2 cells treated with 0.4 mM . We also found that Sirt1 was an upstream regulator of AMPK in hepatocytes. A novel finding of this study was the observation that expression of GLP-1R is proportional to exendin-4 concentration and exendin-4 could attenuate fatty liver through activation of Sirt1.

Keyword: lipogenesis

The fatty -rich fraction of Eruca sativa (rocket salad) leaf extract exerts antidiabetic effects in cultured skeletal muscle, adipocytes and liver cells.

Eruca sativa Mill. (Brassicaceae), commonly known as rocket salad, is a popular leafy-green vegetable with many health benefits.To evaluate the antidiabetic activities of this plant in major insulin-responsive tissues.Five E. sativa leaf extracts of varying polarity were prepared (aqueous extract, 70% and 95% ethanol extracts, the n-hexane-soluble fraction of the 95% ethanol extract (ES3) and the defatted 95% ethanol extract). Eruca sativa extracts were investigated through a variety of cell-based in vitro bioassays for antidiabetic activities in C2C12 skeletal muscle cells, H4IIE hepatocytes and 3T3-L1 adipocytes. Guided by the results of these bioassays, ES3 was fractionated into the saponifiable (SM) and the unspaonifiable (USM) fractions. Glucose uptake was measured using [H]-deoxy-glucose, while the effects on hepatic glucose-6-phosphatase (G6Pase) and were assessed using Wako AutoKit Glucose and AdipoRed assays, respectively.ES3 and its SM fraction significantly stimulated glucose uptake with EC values of 8.0 and 5.8\u2009μg/mL, respectively. Both extracts significantly inhibited G6Pase activity (IC values of 4.8 and 9.3\u2009μg/mL, respectively). Moreover, ES3 and SM showed significant adipogenic activities with EC of 4.3 and 6.1\u2009μg/mL, respectively. Fatty content of SM was identified by GC-MS. trans-Vaccenic and palmitoleic acids were the major unsaturated fatty acids, while and azelaic acids were the main saturated fatty acids.These findings indicate that ES3 and its fatty -rich fraction exhibit antidiabetic activities in insulin-responsive cell lines and may hence prove useful for the treatment of type 2 diabetes.

Keyword: lipogenesis

ToF-SIMS study of differentiation of human bone-derived stromal cells: new insights into osteoporosis.

Lipids have numerous important functions in the human body, as they form the cells\' plasma membranes and play a key role in many disease states, presumably also in osteoporosis. Here, the fatty composition of the outer plasma membranes of cells differentiated into the osteogenic and adipogenic direction is studied with surface-sensitive time-of-flight secondary ion mass spectrometry (ToF-SIMS). For data evaluation, principal component analysis (PCA) is applied. Human (bone-derived) mesenchymal stromal cells (hMSCs) from an osteoporotic donor and a control donor are compared to reveal differences in the fatty composition of the membranes. The chemical information is correlated to staining and real-time quantitative polymerase chain reaction (rt-qPCR) results to provide insight into the gene expression of several differentiation markers on the RNA level. Adipogenic differentiation of hMSCs from a non-osteoporotic donor correlates with increased relative intensities of all fatty acids under investigation. After osteogenic differentiation of non-osteoporotic cells, the relative mass signal intensities of unsaturated fatty acids such as oleic and linoleic acids are increased. However, the osteoporotic cells show increased levels of in the plasma membrane after exposure to osteogenic differentiation conditions, which correlates to an immature differentiation state relative to non-osteoporotic osteogenic cells. This immature differentiation state is confirmed by increased early osteogenic differentiation factor Runx2 on RNA level and by less calcium mineralization spots seen in von Kossa staining and ToF-SIMS images. Graphical abstract Time-of-flight secondary ion mass spectrometry is applied to analyze the fatty composition of the outer plasma membranes of cells differentiated into the adipogenic and osteogenic direction. Cells from an osteoporotic and a control donor are compared to reveal differences due to differentiation and disease stage of the cells.

Keyword: lipogenesis

Deletion of steroid receptor coactivator-3 gene ameliorates hepatic steatosis.

Excess dietary fat can cause hepatic steatosis, which can progress into severe liver disorders including steatohepatitis and cirrhosis. Steroid receptor coactivator-3 (SRC-3), a member of the p160 coactivator family, is reported as a key regulator of and energy homeostasis. We sought to determine the influence of SRC-3 on hepatic steatosis and the mechanism beneath.The influence of siRNA-mediated SRC-3 silencing on hepatic lipid accumulation was assessed in HepG2 cells. The molecular mechanism of SRC-3 regulation of hepatic lipid metabolism was also studied. Moreover, the effect of SRC-3 ablation on hepatic steatosis was examined in SRC-3 deficient mice.In this study, we report that SRC-3 ablation reduces -induced lipid accumulation in HepG2 cells. Moreover, deletion of SRC-3 ameliorates hepatic steatosis and inflammation response in mice fed a high fat diet (HFD). These metabolic improvements can presumably be explained by the reduction in chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) expression and the subsequent elevation in peroxisome proliferator-activated receptor α (PPARα) level. At the molecular level, SRC-3 interacts with retinoic receptor α (RARα) to activate COUP-TFII expression under all-trans retinoic (ARTA) treatment.These findings indicate a crucial role for SRC-3 in regulating hepatic lipid metabolism and provide the possible novel inner mechanisms.Copyright © 2011. Published by Elsevier B.V.

Keyword: lipogenesis

Loss of toll-like receptor 3 aggravates hepatic inflammation but ameliorates steatosis in mice.

The importance of toll-like receptor (TLR) 4 in the pathogenesis of steatohepatitis has been well documented; however, little is known about the role of TLR3. In this study, we determined whether the depletion of TLR3 modulated hepatic injury in mice and further aimed to provide mechanistic insights into the TLR3-mediated modulation of diet-induced hepatic inflammation and fat accumulation. Hepatic steatosis and inflammatory response were induced by feeding wild-type (WT) or TLR3 knockout mice a high-fat diet for 8 weeks. Primary liver resident cells, including hepatocytes, Kupffer cells, and hepatic stellate cells (HSCs), were treated with . TLR3 knockout mice fed a high-fat diet showed severe hepatic inflammation accompanied by nuclear factor-κB and IRF3 activation, which is mainly induced by the activation of Kupffer cells. Decreased TLR4 expression was restored in hepatic mononuclear cells and Kupffer cells in TLR3 knockout mice compared to that in the WT. Moreover, hepatic steatosis was decreased in TLR3 knockout mice. Hepatocytes from TLR3 knockout mice exhibited reduced expression of cannabinoid receptors. HSCs from TLR3 knockout mice showed decreased expression of the enzymes involved in endocannabinoid synthesis. In conclusion, this study suggests that the selective modulation of TLR3 could be a novel therapeutic target for the treatment of hepatic inflammation and steatosis.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Tannic , a novel histone acetyltransferase inhibitor, prevents non-alcoholic fatty liver disease both in\xa0vivo and in\xa0vitro model.

We examined the potential of tannic (TA) as a novel histone acetyltransferase inhibitor (HATi) and demonstrated that TA prevents non-alcoholic fatty liver disease (NAFLD) by inhibiting HAT activity.The anti-HAT activity of TA was examined using HAT activity assays. An in\xa0vitro NAFLD model was generated by treating HepG2 cells with oleic and acids. Male C57BL/6J mice were fed a control diet (CD) or Western diet (WD) with or without supplementation with either 1% or 3% TA (w/w) for 12 weeks. Finally, the possibility of interacting p300 and TA was simulated.TA suppressed HAT activity both in\xa0vitro and in\xa0vivo. Interestingly, TA abrogated occupancy of p300 on the sterol regulatory element in the fatty synthase and ATP-citrate lyase promoters, eventually inducing hypoacetylation of H3K9 and H3K36. Furthermore, TA decreased acetylation at lysine residues 9 and 36 of histone H3 protein and that of total proteins. Consequently, TA decreased the mRNA expression of lipogenesis-related genes and attenuated lipid accumulation in\xa0vivo. We observed that NAFLD features, including body weight, liver mass, fat mass, and lipid profile in serum, were improved by TA supplementation in\xa0vivo. Finally, we demonstrated the possibility that TA directly binds to p300 through docking simulation between ligand and protein.Our findings demonstrate that TA, a novel HATi, has potential application for the prevention of NAFLD.Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: lipogenesis

Polymorphisms in lipogenic genes and milk fatty composition in Holstein dairy cattle.

Changing bovine milk fatty (FA) composition through selection can decrease saturated FA (SFA) consumption, improve human health and provide a means for manipulating processing properties of milk. Our study determined associations between milk FA composition and genes from triacylglycerol (TAG) biosynthesis pathway. The GC dinucleotide allele of diacylglycerol O-acyltransferase 1:g.10433-10434AA >GC was associated with lower (16:0) concentration but higher oleic (18:1 cis-9), linoleic (18:2 cis-9, cis-12) concentrations, and elongation index. Accordingly, the GC dinucleotide allele was associated with lower milk fat percentage and SFA concentrations but higher monounsaturated FA and polyunsaturated FA (PUFA) concentrations. The glycerol-3-phosphate acyltransferase, mitochondrial haplotypes were associated with higher myristoleic (14:1 cis-9) concentration and C14 desaturation index. The 1-acylglycerol-3-phosphate acyltransferase 1 haplotypes were associated with higher PUFA and linoleic concentrations. The results of this study provide information for developing genetic tools to modify milk FA composition in dairy cattle.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Knockdown of triglyceride synthesis does not enhance palmitate lipotoxicity or prevent oleate-mediated rescue in rat hepatocytes.

Experiments in a variety of cell types, including hepatocytes, consistently demonstrate the acutely lipotoxic effects of saturated fatty acids, such as palmitate (PA), but not unsaturated fatty acids, such as oleate (OA). PA+OA co-treatment fully prevents PA lipotoxicity through mechanisms that are not well defined but which have been previously attributed to more efficient esterification and sequestration of PA into triglycerides (TGs) when OA is abundant. However, this hypothesis has never been directly tested by experimentally modulating the relative partitioning of PA/OA between TGs and other lipid fates in hepatocytes. In this study, we found that addition of OA to PA-treated hepatocytes enhanced TG synthesis, reduced total PA uptake and PA lipid incorporation, decreased phospholipid saturation and rescued PA-induced ER stress and lipoapoptosis. Knockdown of diacylglycerol acyltransferase (DGAT), the rate-limiting step in TG synthesis, significantly reduced TG accumulation without impairing OA-mediated rescue of PA lipotoxicity. In both wild-type and DGAT-knockdown hepatocytes, OA co-treatment significantly reduced PA lipid incorporation and overall phospholipid saturation compared to PA-treated hepatocytes. These data indicate that OA\'s protective effects do not require increased conversion of PA into inert TGs, but instead may be due to OA\'s ability to compete against PA for cellular uptake and/or esterification and, thereby, normalize the composition of cellular lipids in the presence of a toxic PA load.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: lipogenesis

Adipocyte-specific inactivation of Acyl-CoA synthetase fatty transport protein 4 (Fatp4) in mice causes adipose hypertrophy and alterations in metabolism of complex lipids under high fat diet.

Fatp4 exhibits acyl-CoA synthetase activity and is thereby able to catalyze the activation of fatty acids for further metabolism. However, its actual function in most tissues remains unresolved, and its role in cellular fatty uptake is still controversial. To characterize Fatp4 functions in adipocytes in vivo, we generated a mouse line with adipocyte-specific inactivation of the Fatp4 gene (Fatp4(A-/-)). Under standard conditions mutant mice showed no phenotypical aberrance. Uptake of radiolabeled and lignoceric into adipose tissue of Fatp4(A-/-) mice was unchanged. When exposed to a diet enriched in long chain fatty acids, Fatp4(A-/-) mice gained more body weight compared with control mice, although they were not consuming more food. Pronounced obesity was accompanied by a thicker layer of subcutaneous fat and greater adipocyte circumference, although expression of genes involved in de novo was not changed. However, the increase in total fat mass was contrasted by a significant decrease in various phospholipids, sphingomyelin, and cholesteryl esters in adipocytes. Livers of Fatp4-deficient animals under a high fat diet exhibited a higher degree of fatty degeneration. Nonetheless, no evidence for changes in insulin sensitivity and adipose inflammation was found. In summary, the results of this study confirm that Fatp4 is not crucial for fatty uptake into adipocytes. Instead, under the condition of a diet enriched in long chain fatty acids, adipocyte-specific Fatp4 deficiency results in adipose hypertrophy and profound alterations in the metabolism of complex lipids.

Keyword: lipogenesis

The plasma lipidomic signature of nonalcoholic steatohepatitis.

Specific alterations in hepatic lipid composition characterize the spectrum of nonalcoholic fatty liver disease (NAFLD), which extends from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (NASH). However, the plasma lipidome of NAFLD and whether NASH has a distinct plasma lipidomic signature are unknown. A comprehensive analysis of plasma lipids and eicosanoid metabolites quantified by mass spectrometry was performed in NAFL (n = 25) and NASH (n = 50) subjects and compared with lean normal controls (n = 50). The key findings include significantly increased total plasma monounsaturated fatty acids driven by palmitoleic (16:1 n7) and oleic (18:1 n9) acids content (P < 0.01 for both acids in both NAFL and NASH). The levels of palmitoleic , oleic , and palmitoleic to (16:0) ratio were significantly increased in NAFLD across multiple lipid classes. Linoleic (8:2n6) was decreased (P < 0.05), with a concomitant increase in gamma-linolenic (18:3n6) and dihomo gamma-linolenic (20:3n6) acids in both NAFL and NASH (P < 0.001 for most lipid classes). The docosahexanoic (22:6 n3) to docosapentenoic (22:5n3) ratio was significantly decreased within phosphatidylcholine (PC), and phosphatidylethanolamine (PE) pools, which was most marked in NASH subjects (P < 0.01 for PC and P < 0.001 for PE). The total plasmalogen levels were significantly decreased in NASH compared with controls (P < 0.05). A stepwise increase in lipoxygenase (LOX) metabolites 5(S)-hydroxyeicosatetraenoic (5-HETE), 8-HETE, and 15-HETE characterized progression from normal to NAFL to NASH. The level of 11-HETE, a nonenzymatic oxidation product of arachidonic (20:4) , was significantly increased in NASH only.Although increased , desaturases, and LOX activities characterize NAFL and NASH, impaired peroxisomal polyunsaturated fatty (PUFA) metabolism and nonenzymatic oxidation is associated with progression to NASH.

Keyword: lipogenesis

[The evaluation of unsaturation of blood lipids using methods of physical chemistry and clinical biochemistry. The insulin regulation of metabolism of fatty acids, number of double binds and cell absorption of glucose].

It is supposed that the main cause of insulin synthesis at late stages of phylogenesis became discrepancy between increase in vivo need in energy and physical chemical parameters of saturated fatty ; its transportation to cells in composition of lipoproteins in optimal quantity (more than 15% of all fatty acids) became in vivo unfeasible. The biological role of insulin consists in supporting of insulin-dependent cells (skeletal miocytes in the first place) with substrates for gaining energy. The hormone transforms all saturated fatty endogenously synthesized from glucose into specific for animal cells rn-9 C18:1 oleic mono unsaturated fatty . The endogenous mono unsaturated fatty is oxidized by mitohondria with the highest constant of reaction velocity gaining for cells optimal quantity of biotransforming energy in the form of ATP. The insulin expresses in hepatocytes synthesis of oleic triglycerides and formation of oleic lipoproteins of very low density that only insulin-dependent cells absorb using apoE/B-100-endocytosis. The insulin expresses synthesis of Palmitoyl-KoA-elongase, stearyl-KoA-desaturase and glucose transporters 4, activates glucose absorption by cells with the purpose of synthesis endogenous oleic saturated fatty . The insulin substitutes in vivo ineffective alternative of metabolism of fatty acids for potentially more effective oleic metabolism of fatty acids. The insulin increases unsaturation of fatty acids and number of double binds in them. This can be established by direct titration of double binds by ozone on the basis of quantitative detection of fatty acids using technique of gas chromatography and calculating ratio C16:1/C16:0, C18:1/C18:0 and C18:1/C16:0. The diabetes mellitus is a disorder of metabolism of mono unsaturated fatty in the first place and only in the second place pathology of glucose absorption by cells.

Keyword: lipogenesis

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: lipogenesis

Exendin-4 Inhibits Hepatic by Increasing β-Catenin Signaling.

The aim of this study is to investigate whether the beneficial effect of exendin-4 on hepatic steatosis is mediated by β-catenin signaling. After the HepG2 human hepatoma cells were treated with PA for 24 hours, total triglycerides levels were increased in a dose-dependent manner, and the expression levels of perilipin family members were upregulated in cells treated with 400 μM PA. For our in vitro model of hepatic steatosis, HepG2 cells were treated with 400 μM (PA) in the presence or absence of 100 nM exendin-4 for 24 hours. PA increased the expression of lipogenic genes, such as sterol regulatory element-binding protein 1c (SREBP-1c), peroxisome proliferator-activated receptor gamma (PPARγ), stearoyl-CoA desaturase 1 (SCD1), fatty synthase (FAS), and acetyl-CoA carboxylase (ACC) and triglyceride synthesis-involved genes, such as diacylglycerol acyltransferase 1 (DGAT1) and diacylglycerol acyltransferase 2 (DGAT2) in HepG2 cells, whereas exendin-4 treatment significantly prevented the upregulation of SREBP-1c, PPARγ, SCD1, FAS, ACC, DGAT1 and DGAT2. Moreover, exendin-4 treatment increased the expression of phosphorylated glycogen synthase kinase-3 beta (GSK-3β) in the cytosolic fraction and the expression of β-catenin and transcription factor 4 (TCF4) in the nuclear fraction. In addition, siRNA-mediated inhibition of β-catenin upregulated the expression of lipogenic transcription factors. The protective effects of exendin-4 on intracellular triglyceride content and total triglyceride levels were not observed in cells treated with the β-catenin inhibitor IWR-1. These data suggest that exendin-4 treatment improves hepatic steatosis by inhibiting via activation of Wnt/β-catenin signaling.

Keyword: lipogenesis

Stereospecificity of fatty 2-hydroxylase and differential functions of 2-hydroxy fatty enantiomers.

FA 2-hydroxylase (FA2H) is an NAD(P)H-dependent enzyme that initiates FA α oxidation and is also responsible for the biosynthesis of 2-hydroxy FA (2-OH FA)-containing sphingolipids in mammalian cells. The 2-OH FA is chiral due to the asymmetric carbon bearing the hydroxyl group. Our current study performed stereochemistry investigation and showed that FA2H is stereospecific for the production of (R)-enantiomers. FA2H knockdown in adipocytes increases diffusional mobility of raft-associated lipids, leading to reduced GLUT4 protein level, glucose uptake, and . The effects caused by FA2H knockdown were reversed by treatment with exogenous (R)-2-hydroxy , but not with the (S)-enantiomer. Further analysis of sphingolipids demonstrated that the (R)-enantiomer is enriched in hexosylceramide whereas the (S)-enantiomer is preferentially incorporated into ceramide, suggesting that the observed differential effects are in part due to synthesis of sphingolipids containing different 2-OH FA enantiomers. These results may help clarify the mechanisms underlying the recently identified diseases associated with FA2H mutations in humans and may lead to potential pharmaceutical and dietary treatments. This study also provides critical information to help study functions of 2-OH FA enantiomers in FA α oxidation and possibly other sphingolipid-independent pathways.

Keyword: lipogenesis

CHP1 Regulates Compartmentalized Glycerolipid Synthesis by Activating GPAT4.

Cells require a constant supply of fatty acids to survive and proliferate. Fatty acids incorporate into membrane and storage glycerolipids through a series of endoplasmic reticulum (ER) enzymes, but how these enzymes are regulated is not well understood. Here, using a combination of CRISPR-based genetic screens and unbiased lipidomics, we identified calcineurin B homologous protein 1 (CHP1) as a major regulator of ER glycerolipid synthesis. Loss of\xa0CHP1 severely reduces fatty incorporation and storage in mammalian cells and invertebrates. Mechanistically, CHP1 binds and activates GPAT4, which catalyzes the initial rate-limiting step in glycerolipid synthesis. GPAT4 activity requires CHP1 to be N-myristoylated, forming a key molecular interface between the two proteins. Interestingly, upon CHP1 loss, the peroxisomal enzyme, GNPAT, partially compensates for the loss of ER lipid synthesis, enabling cell proliferation. Thus, our work identifies a conserved regulator of glycerolipid metabolism and reveals plasticity in lipid synthesis of proliferating cells.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Reductive glutamine metabolism by IDH1 mediates under hypoxia.

Acetyl coenzyme A (AcCoA) is the central biosynthetic precursor for fatty- synthesis and protein acetylation. In the conventional view of mammalian cell metabolism, AcCoA is primarily generated from glucose-derived pyruvate through the citrate shuttle and ATP citrate lyase in the cytosol. However, proliferating cells that exhibit aerobic glycolysis and those exposed to hypoxia convert glucose to lactate at near-stoichiometric levels, directing glucose carbon away from the tricarboxylic cycle and fatty- synthesis. Although glutamine is consumed at levels exceeding that required for nitrogen biosynthesis, the regulation and use of glutamine metabolism in hypoxic cells is not well understood. Here we show that human cells use reductive metabolism of α-ketoglutarate to synthesize AcCoA for lipid synthesis. This isocitrate dehydrogenase-1 (IDH1)-dependent pathway is active in most cell lines under normal culture conditions, but cells grown under hypoxia rely almost exclusively on the reductive carboxylation of glutamine-derived α-ketoglutarate for de novo . Furthermore, renal cell lines deficient in the von Hippel-Lindau tumour suppressor protein preferentially use reductive glutamine metabolism for lipid biosynthesis even at normal oxygen levels. These results identify a critical role for oxygen in regulating carbon use to produce AcCoA and support lipid synthesis in mammalian cells.

Keyword: lipogenesis

High-Protein Intake during Weight Loss Therapy Eliminates the Weight-Loss-Induced Improvement in Insulin Action in Obese Postmenopausal Women.

High-protein (HP) intake during weight loss (WL) therapy is often recommended because it reduces the loss of lean tissue mass. However, HP intake could\xa0have adverse effects on metabolic function, because\xa0protein ingestion reduces postprandial insulin sensitivity. In this study, we compared the effects of ∼10% WL with a hypocaloric diet containing 0.8\xa0g protein/kg/day and a hypocaloric diet containing 1.2\xa0g protein/kg/day on muscle insulin action in\xa0postmenopausal women with obesity. We found that HP intake reduced the WL-induced decline in lean tissue mass by ∼45%. However, HP intake also prevented the WL-induced improvements in muscle insulin signaling and insulin-stimulated glucose uptake, as well as the WL-induced adaptations in oxidative stress and cell structural biology pathways. Our data demonstrate that the protein content of a WL diet can have profound effects on metabolic function and underscore the importance of considering dietary macronutrient composition during WL therapy for people with obesity.Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

Accumulation of lipids and oxidatively damaged DNA in hepatocytes exposed to particles.

Exposure to particles has been suggested to generate hepatosteatosis by oxidative stress mechanisms. We investigated lipid accumulation in cultured human hepatocytes (HepG2) and rat liver after exposure to four different carbon-based particles. HepG2 cells were exposed to particles for 3h and subsequently incubated for another 18h to manifest lipid accumulation. In an animal model of metabolic syndrome we investigated the association between intake of carbon black (CB, 14nm) particles and hepatic lipid accumulation, inflammation and gene expression of Srebp-1, Fasn and Scd-1 involved in lipid synthesis. There was a concentration-dependent increase in intracellular lipid content after exposure to CB in HepG2 cells, which was only observed after co-exposure to oleic/. Similar results were observed in HepG2 cells after exposure to diesel exhaust particles, fullerenes C60 or pristine single-walled carbon nanotubes. All four types of particles also generated oxidatively damaged DNA, assessed as formamidopyrimidine DNA glycosylase (FPG) sensitive sites, in HepG2 cells after 3h exposure. The animal model of metabolic syndrome showed increased lipid load in the liver after one oral exposure to 6.4mg/kg of CB in lean Zucker rats. This was not associated with increased iNOS staining in the liver, indicating that the oral CB exposure was associated with hepatic steatosis rather than steatohepatitis. The lipid accumulation did not seem to be related to increased because there were unaltered gene expression levels in both the HepG2 cells and rat livers. Collectively, exposure to particles is associated with oxidative stress and steatosis in hepatocytes.© 2013. Published by Elsevier Inc. All rights reserved.

Keyword: lipogenesis

miR-1224-5p Enhances Hepatic by Targeting Adenosine Monophosphate-Activated Protein Kinase α1 in Male Mice.

MicroRNAs are potential therapeutic targets for metabolic diseases. Here, miR-1224-5p was highly expressed in the livers of mice fed a high-fat diet (HFD) and in obese (ob/ob) mice. To examine the potential role of miR-1224-5p, we constructed liver-specific adenoviral vectors expressing either an miR-1224-5p inhibitor sequence or miR-1224-5p mimic sequences. After tail-vein vector injection, HFD-fed mice were examined for expression of lipogenic genes. We found that miR-1224-5p inhibitors significantly attenuated hepatic and steatosis in HFD-fed mice, whereas miR-1224-5p mimicked promoted lipid accumulation in the liver of chow-fed C57BL/6 mice. Additional in vitro studies demonstrated that downregulation of miR-1224-5p in HepG2 and primary hepatocytes led to a reduction of cellular triglycerides after treatment with an oleic and mixture. Importantly, this study also identified adenosine monophosphate-activated protein kinase (AMPK)-α1 as a direct target of miR-1224-5p. miR-1224-5p binding to the 3\' untranslated region of AMPKα1 suppressed expression of the AMPKα1 protein and its downstream molecules. Metformin, an activator of AMPK, also inhibited hepatic expression of miR-1224-5p. Together, these findings indicate that miR-1224-5p promotes hepatic by suppressing AMPKα1 expression and suggest that miR-1224-5p inhibitors warrant further investigation as potential therapeutic tools in the treatment of nonalcoholic fatty liver disease.

Keyword: lipogenesis

Alpha-mangostin induces endoplasmic reticulum stress and autophagy which count against fatty synthase inhibition mediated apoptosis in human breast cancer cells.

One of the most important metabolic hallmarks of breast cancer cells is enhanced . Increasing evidences suggest that fatty synthase (FAS) plays an important role in human breast cancer. Previously we discovered that alpha-mangostin showed apoptotic effect on human breast cancer cells via inhibiting FAS activity. The endoplasmic reticulum (ER) stress and autophagy are involved in cell apoptosis. However, the role of ER stress and autophagy in FAS inhibition induced apoptosis still remains unclear.We evaluated the effects of alpha-mangostin on ER stress and autophagy in human breast cancer cells. Intracellular FAS activity was measured by a spectrophotometer at 340\xa0nm of NADPH absorption. Cell Counting Kit assay was used to test the cell viability. Immunoblot analysis was performed to detect protein expression levels. Apoptotic effects were detected by flow cytometry.Alpha-mangostin induced endoplasmic reticulum stress and autophagy, both of which reduced the apoptotic effect of alpha-mangostin in MDA-MB-231 cells. , the end product of FAS catalyzed reaction, rescued the ER stress and autophagy induced by alpha-mangostin. Cell apoptosis was markedly promoted by inhibiting ER stress and autophagy while treating cells with alpha-mangostin.We propose a hypothesis that a combination of FAS inhibition and ER stress and autophagy inhibition has an application potential in the chemoprevention and treatment of breast cancer.

Keyword: lipogenesis

Adipokines secretion in feline primary adipose tissue culture in response to dietary fatty acids.

Obesity in cats has been associated with alterations in adipokines including: adiponectin, interleukin-6 (IL6), and tumor necrosis factor-α (TNFα). Omega-3 polyunsaturated fatty acids have multiple beneficial effects on obesity-associated disorders, and therefore may alleviate these alterations. This study aimed to determine the effects of body condition, fat depot, troglitazone, and different fatty acids on secretion of adiponectin, IL6 and TNFα from adipose tissue of healthy cats. Subcutaneous and visceral adipose tissue samples were collected from 18 healthy intact female cats, and body condition score (Range 3-7/9) was determined. Concentrations of adiponectin were measured in mature adipocytes cultures and concentrations of IL6 and TNFα were measured in stromovascular cells cultures following treatment with control medium, troglitazone at 10\u2009μM, eicosapentaenoic , arachidonic , or , at 25, 50, or 100\u2009μM.Stromovascular cells of visceral origin secreted higher concentrations of IL6 than corresponding cells of subcutaneous origin (P\u2009=\u20090.003). Arachidonic treatment at 25, 50, and 100\u2009μM increased IL6 secretion in subcutaneous (P\u2009=\u20090.045, P\u2009=\u20090.002, and P\u2009<\u20090.001, respectively) and visceral (P\u2009=\u20090.034, P\u2009=\u20090.001, and P\u2009<\u20090.001, respectively) stromovascular cells. Eicosapentaenoic treatment increased TNFα secretion in subcutaneous stromovascular cells at 25, 50, and 100\u2009μM (P\u2009=\u20090.002, P\u2009=\u20090.001, and P\u2009=\u20090.015, respectively) and in visceral stromovascular cells at 50\u2009μM (P\u2009<\u20090.001). No significant effect on medium adiponectin concentration was observed following troglitazone treatment (P\u2009=\u20090.4) or fatty acids treatments at 25 (P\u2009=\u20090.2), 50 (P\u2009=\u20090.8), or 100 (P\u2009=\u20090.7) μM. Body condition score did not have significant effects on medium concentrations of adiponectin (P\u2009=\u20090.4), IL6 (P\u2009=\u20090.1), or TNFα (P\u2009=\u20090.8).This study demonstrated higher basal secretion of IL6 from visceral compared to subcutaneous adipose tissue, a stimulatory effect of arachidonic on secretion of IL6 and a stimulatory effect of eicosapentaenoic on TNFα from feline adipose tissue.

Keyword: lipogenesis

Development of an in vitro model to study hepatitis C virus effects on hepatocellular lipotoxicity and lipid metabolism.

Hepatic steatosis is common in patients infected with hepatitis C virus (HCV). Particularly in patients infected with non-genotype 3 HCV, hepatic steatosis is closely related to factors of the such as hyperlipidemia. However, the molecular mechanisms involved in this "" steatosis in non-3 genotype HCV infections are not well understood. Here, we aimed to develop an in vitro model to study the effect of genotype 1 HCV infection on hepatic lipotoxicity and lipid metabolism. Cellular lipid accumulation was induced in Huh-7 hepatoma cells transfected with HCV genotype 1b replicon (HCV) by incubation with increasing doses of (C16:0) or oleic (C18:1 n-9) complexed to albumin mimicking hyperlipidemic conditions. Mock transfected hepatoma cells (HCV) were used as controls. Incubation with oleic concentrations as high as 0.5\u202fmM did not induce toxic effects in HCV or HCV cells. In contrast, incubation with caused dose-dependently cytotoxic effects which were more pronounced in HCV compared to HCV cells. Further analysis with subtoxic and oleic concentrations revealed a higher uptake of fatty acids and intracellular triglyceride accumulation in HCV compared to HCV cells. Carnitine palmitoyltransferase I (CPT1) expression, indicative of mitochondrial beta-oxidation, was markedly stimulated by lipid exposure in HCV but not in HCV cells. Furthermore, heme oxygenase 1 (HMOX1) expression levels increased in FA stimulated cells, and this increase was significantly higher in HCV compared to HCV cells. In contrast, expression of the key enzymes of hepatic de novo lipogenesis fatty synthase (FASN) and stearoyl-CoA desaturase (SCD-1) was significantly reduced upon oleate exposure in HCV but not in HCV cells. In summary, our newly developed cell culture model revealed effects of HCV genotype 1b infection on susceptibility to lipid accumulation and toxicity particularly to saturated lipids. These results may indicate that HCV (genotype 1b) infected individuals with hyperlipidemia may benefit from dietary or pharmacological intervention.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: metabolic syndrome

Two Cockayne patients with a novel splice site mutation - clinical and analyses.

Cockayne (CS) is a rare autosomal recessive disorder, which leads to neurodegeneration, growth failure and premature aging. Most of the cases are due to mutations in the ERCC6 gene, which encodes the protein CSB. CSB is involved in several functions including DNA repair and transcription. Here we describe two Danish brothers with CS. Both patients carried a novel splice site mutation (c.2382+2T>G), and a previously described nonsense mutation (c.3259C>T, p.Arg1087X) in a biallelic state. Both patients presented the cardinal features of the disease including microcephaly, congenital cataract and postnatal growth failure. In addition, their fibroblasts were hypersensitive to UV irradiation and exhibited increased superoxide levels in comparison to fibroblasts from healthy age and gender matched individuals. Metabolomic analysis revealed a distinctive profile in cells from the CS patients compared to control cells. Among others, α-ketoglutarate, hydroxyglutarate and certain amino acids (ornithine, proline and glycine) were reduced in the CS patient fibroblasts, whereas glycolytic intermediates (glucose-6-phosphate and pyruvic ) and fatty acids (, stearic and myristic ) were increased. Our data not only provide additional information to the database of CS mutations, but also point towards targets for potential treatment of this devastating disease.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Reduced circulating levels of sTWEAK are associated with NAFLD and may affect hepatocyte triglyceride accumulation.

Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the and is strongly associated with obesity, dyslipidaemia and altered glucose regulation. Previous data demonstrated that low circulating levels of tumour necrosis factor weak inducer of apoptosis (sTWEAK) were associated with obesity, diabetes and insulin resistance, all traits associated with an increased risk of NALFD. Circulating sTWEAK levels are expected to be reduced in the presence of NAFLD.We aimed to explore the relationship between NAFLD and circulating sTWEAK levels in obese patients, and to evaluate the effect of sTWEAK on hepatocyte triglyceride accumulation.Design setting and patients:This is an observational case-control study performed in n=112 severely obese patients evaluated for NAFLD by abdominal ultrasound and n=32 non-obese patients without steatosis. Serum sTWEAK concentrations were measured by ELISA. Multivariable analyses were performed to determine the independent predictors of NAFLD. We analysed TWEAK and Fn14 protein expression in liver biopsies by western blotting and immunohistochemistry. An immortalized primary human hepatocyte cell line (HHL) was used to evaluate the effect of sTWEAK on triglyceride accumulation.We observed a reduction in serum circulating sTWEAK concentrations with the presence of liver steatosis. On multivariable analysis, lower sTWEAK concentrations were independently associated with the presence of NAFLD (odds ratio (OR)=0.023; 95% confidence interval: 0.001-0.579; P<0.022). In human hepatocytes, sTWEAK administration reduced fat accumulation as demonstrated by the reduction in -induced accumulation of triglyceride and the decreased expression of cluster of differentiation 36 (CD36) and perilipin 1 and 2 (PLIN1 and PLIN2) genes.Decreased sTWEAK concentrations are independently associated with the presence of NAFLD. This is concordant with the observation that TWEAK reduces lipid accumulation in human liver cells.

Keyword: metabolic syndrome

ICER-1gamma overexpression drives palmitate-mediated connexin36 down-regulation in insulin-secreting cells.

Channels formed by the gap junction protein connexin36 (Cx36) contribute to the proper control of insulin secretion. We investigated the impact of chronic hyperlipidemia on Cx36 expression in pancreatic beta-cells. Prolonged exposure to the saturated free fatty palmitate reduced the expression of Cx36 in several insulin-secreting cell lines and isolated mouse islets. The effect of palmitate was fully blocked upon protein kinase A (PKA) inhibition by H89 and (Rp)-cAMP, indicating that the cAMP/PKA pathway is involved in the control of Cx36 expression. Palmitate treatment led to overexpression of the inducible cAMP early repressor (ICER-1gamma), which bound to a functional cAMP-response element located in the promoter of the CX36 gene. Inhibition of ICER-1gamma overexpression prevented the Cx36 decrease, as well as the palmitate-induced beta-cell secretory dysfunction. Finally, freshly isolated islets from mice undergoing a long term high fat diet expressed reduced Cx36 levels and increased ICER-1gamma levels. Taken together, these data demonstrate that chronic exposure to palmitate inhibits the Cx36 expression through PKA-mediated ICER-1gamma overexpression. This Cx36 down-regulation may contribute to the reduced glucose sensitivity and altered insulin secretion observed during the pre-diabetic stage and in the .

Keyword: metabolic syndrome

Serum fatty acids in postinfarction middle-aged men.

There is a multitude of data showing that coronary heart disease is affected by the quality of dietary fat. The fatty composition of serum lipids has been shown to reflect that of the diet. It is likely that, after myocardial infarction, both the health-care professionals and the patients themselves pay more attention to dietary guidelines. In order to assess the correctness of this assumption, we compared the composition of serum fatty acids in 40 male subjects with a history of myocardial infarction (MI) with that of 40 age-matched controls, both from the FINRISK study. The percentage composition of fatty acids of total serum lipids was analysed by gas chromatography. In comparison with the control group, the MI group had higher body mass index (BMI), a higher prevalence of diabetes, higher level of serum triglycerides and a lower level of serum high-density lipoprotein (HDL) cholesterol, all indicators of the . The MI group had higher proportions of serum (16:0) and oleic acids (18:1), and a lower proportion of linoleic (18:2 n-6) than the control group. The is accompanied by an elevated level of serum insulin, which is known to enhance the synthesis of saturated and monounsaturated fatty acids, such as 16:0 and 18:1, and to stimulate the activity delta-6 desaturase, decreasing the concentration of linoleic . Our results suggest that the observed serum fatty composition in subjects with coronary heart disease is dependent on factors in addition to dietary fatty composition.

Keyword: metabolic syndrome

Development of insulin resistance in the JCR:LA-cp rat: role of triacylglycerols and effects of MEDICA 16.

The JCR:LA-cp rat develops an extreme obese/insulin-resistant such that by 12 weeks of age, there is no longer any insulin-mediated glucose turnover. At 4 weeks of age, obese and lean rats have essentially identical basal and insulin-mediated glucose uptake in skeletal muscle. By 8 weeks of age, however, the obese rats no longer exhibit such intake. Plasma insulin concentrations in the normal fed state show only small increases up to 4 weeks, with a rapid rise to a marked hyperinsulinemia thereafter, with an age at half-development of 5.5 weeks. Plasma triacylglycerol concentrations in fed obese rats are elevated at 3 weeks and rise rapidly thereafter. The triacylglycerol content of skeletal muscle is significantly elevated in the obese rats at 4 weeks of age. Histological examination of Oil Red O-stained muscle tissue and transmission electron microscopy shows the presence of intracellular lipid droplets. Treatment with the potent triacylglycerol-lowering agent MEDICA 16 (beta,beta\'-tetramethylhexadecanedioic ) from 6 weeks of age reduces plasma lipids markedly, but it reduces body weight and insulin resistance only modestly. In contrast, treatment with MEDICA 16 from the time of weaning at 3 weeks of age results in the normalization of food intake and body weight to over 8 weeks of age. The development of hyperinsulinemia is also delayed until 8.5 weeks of age, and insulin levels remain strongly reduced. Plasma triacylglycerol concentrations remain at the same level as in lean rats, and neither an elevated muscle triacylglycerol content nor intracellular lipid droplets are found at 4 weeks of age. The results indicate that insulin resistance develops in the young animals and is not directly due to a genetically determined defect in insulin metabolism. The mechanism of induction instead appears to be related to an exaggerated triacylglycerol metabolism.

Keyword: metabolic syndrome

Hepatic TLR4 signaling in obese NAFLD.

Nonalcoholic fatty liver disease occurs frequently in the setting of , but the factors leading to nonalcoholic steatohepatitis (NASH) are not fully understood. This study investigated Toll-like receptor 4 (TLR4) signaling in human liver with the goal of delineating whether activation of this pathway segregates those with nonalcoholic fatty liver from those with NASH. Experiments were performed using liver biopsy tissue obtained from class III obese subjects undergoing bariatric surgery, and extended to an immortalized human hepatocyte HepaRG cell line and primary human hepatocytes. The bacterial endotoxin lipopolysaccharide (LPS) and total free fatty levels were significantly increased in plasma of NASH patients. TLR4 mRNA levels were significantly increased in subjects with NASH compared with NAFL as was interferon regulatory factor (IRF) 3 in the myeloid differentiation factor 88-independent signaling pathway. In HepaRG cells, nuclear factor-κB (NF-κB) nuclear translocation and functional activity increased following treatment with the fatty , palmitate, and following exposure to LPS compared with hepatocytes stimulated with a lipogenic treatment that induced de novo lipogenesis. Palmitate and LPS induction of NF-κB activity was partially attenuated by chemical- or small-interfering RNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was upregulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-κB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition. In conclusion, TLR4 expression is upregulated in a large cohort of NASH patients, compared with those with NAFL, and this occurs within the setting of increased LPS and fatty acids.Copyright © 2015 the American Physiological Society.

Keyword: metabolic syndrome

An environmental wide association study (EWAS) approach to the .

Environmental contaminants have previously been linked to components of the (MetS). However, exposure to environmental contaminants is in part determined by various lifestyle factors.Using an "Environmental Wide Association Study" (ELWAS) integrating environmental contaminants and lifestyle factors, we aimed to evaluate a possible additive role of both contaminants and lifestyle factors regarding MetS.1016 subjects aged 70years were investigated in the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study. 43 environmental contaminants were measured in the circulation. Dietary records were used to evaluate 21 nutrients and the proportions of 13 fatty acids were determined in serum cholesterol esters to further quantify fat quality intake. Adding 5 other important lifestyle factors yielded together 76 environmental and lifestyle factors. MetS was defined by the NCEP/ATPIII-criteria.23% had MetS. Using cross-validation within the sample, fourteen environmental contaminants or lifestyle factors consistently showed a false discovery rate <0.05. When the major variables entered a multiple model, only p, p\'-DDE levels (positive), PCB209 (inverse) and exercise habits (inverse) were together with a fatty pattern, with high levels of and oleic and low levels of linoleic , related to MetS (p<0.002 for all variables).Using a cross-sectional EWAS approach, certain environmental contaminants and lifestyle factors were found to be associated with prevalent in an additive fashion in an elderly population.Copyright © 2013 Elsevier Ltd. All rights reserved.

Keyword: metabolic syndrome

Ceramide mediates inhibition of the AKT/eNOS signaling pathway by palmitate in human vascular endothelial cells.

In , down-regulation of the insulin signaling leads to insulin-regulated metabolism and cardiovascular dyfunctions. Free fatty acids (FFAs) in the circulation are increased in this disorder and inhibit insulin signaling. Lipid oversupply contributes to the development of insulin resistance, likely by promoting the accumulation of lipid metabolites capable of inhibiting signal transduction.This study was designed to examine the effects of FFAs and their metabolites on the insulin signaling pathway that leads to the activation of endothelial nitric oxide synthase (eNOS) and increase in nitric oxide (NO) production in endothelial cells.Here we demonstrate that exposing human umbilical vein endothelial cells (HUVECs) to palmitate inhibits activation of Akt/eNOS signal pathway by insulin, and subsequently insulin-stimulated NO generation. Palmitate concomitantly induced the accumulation of ceramide, a product of acyl-CoA that has been shown to accumulate in insulin-resistant tissues and to inhibit insulin signaling. Preventing de novo ceramide synthesis abolished the antagonistic effect of palmitate toward the Akt/ eNOS pathway. Moreover, inducing ceramide buildup augmented the inhibitory effect of palmitate.Taken together, we have demonstrated that induces accumulation of ceramide, which appears to mediate \'s inhibitory effects on the Akt/eNOS pathway, leading to a significant decrease in NO generation. Therefore, ceramide is a necessary and sufficient intermediate mediating the inhibition of the AKT/eNOS signaling pathway by palmitate in endothelial cells.

Keyword: metabolic syndrome

[The insulin initiates "kinetic perfection" of biologic function of locomotion. The glucose as substratum for synthesis of ω-9 oleic fatty by cross-striated miocytes.]

The phylogenesis theory affords ground for the following propositions. 1. There is no absorption of glucose from intercellular medium by cells in vivo until there is possibility to absorb polar fatty acids from associates with albumin. 2. The late in phylogenesis humoral insulin regulates no stages of glucose metabolism; they are completed a billion years before hormone synthesis. 3. The phylogenetically late insulin is "hostage" of biological function of trophology, function of nutrition, biological reaction of exotrophy; it has no possibility to decrease in food excessed amount of physiologic saturated fatty acids with low kinetic parameters of β-oxidation in mitochondria. 4. The early in phylogenesis, resistant organizational to insulin pool of visceral fatty cells of omentum and late pool of insulin-dependent adipocytes are different in many functional parameters. 5. All " pandemics" such as of resistance to insulin, atherosclerosis, arterial hypertension, and obesity are primarily pathologies of fatty acids. 6. All " pandemics" are pathologies of one biological function, function of locomotion under single algorithm of formation of their pathogenesis. 7. The etiological factor of " pandemics" is uniform - effect of environmental factors in form of disorder of biological function of trophology, function of nutrition; aphysiological excess content in food of saturated fatty , aphysiological trans-forms of fatty acids and ω-7-palmitoleic mono unsaturated fatty . The insulin activates absorption by myocytes, cardiomyocytes of glucose as substrate of synthesis out of it in situ de novo ω-9 oleic mono unsaturated fatty . With such physical chemical parameters that mitochondria oxidize it with the most high constant of velocity of reaction and high effectiveness of formation of ATP.

Keyword: metabolic syndrome

Palmitate induces C-reactive protein expression in human aortic endothelial cells. Relevance to fatty -induced endothelial dysfunction.

Circulating levels of free fatty acids are commonly elevated in patients with the and exert, through activating proinflammatory pathways, harmful effects of the vascular endothelium. In this study, we examined the effect of palmitate (PA) on endothelial C-reactive protein (CRP) expression and the role of CRP in PA-induced nitric oxide (NO) inhibition. Palmitate increased, in a dose-dependent manner, CRP protein expression and production in human aortic endothelial cells (HAECs). Induction of CRP protein was mimicked by ceramide, whereas bromopalmitate and other common free fatty acids such as oleate or linoleate were ineffective. Palmitate also elicited reactive oxygen species production in HAECs, an effect prevented by protein kinase C (PKC) inhibition and adenosine monophosphate-activated kinase (AMPK) activation. Palmitate-treated HAECs showed increased CRP messenger RNA expression and nuclear factor (NF)-κB activation. Induction of CRP expression by PA was prevented by antioxidants and normalized by PKC and mitogen-activated protein kinase inhibitors. Disrupting NF-κB and Janus kinase/signal transducers and activators of transcription pathways or inducing AMPK activation also suppressed the stimulatory effect of PA on CRP messenger RNA expression. Finally, in HAECs, PA reduced NO release, an effect reversed by anti-CRP antibody. These data demonstrate that PA-induced endothelial CRP expression involves PKC-driven oxidative stress, possibly through AMPK inhibition, and activation of downstream redox-sensitive signaling pathways, including NF-κB. They further support a role for endothelial cell-derived CRP as mediator of the suppressive effect of PA on NO production.Copyright © 2011 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

[Composition of the nonesterified fatty acids and lipid peroxidation in ].

Composition of the nonesterified fatty acids in plasma in patients and in other syndromes of insulin resistance is altered. Fatty profile in plasma is related to the composition of dietary fat and to the changes of fatty acids, e.g. to de novo lipogenesis, beta-oxidation and conversion accompanying the oxidative stress. The aim of the work was to study the fatty composition in the major plasma lipid classes in relation to the insulin resistance, to some polymorphisms of candidate genes with activity related to insulin resistance, and to the lipoprotein composition and parameters of lipid peroxidation.95 patients with (56 M/39 F) and 195 healthy persons (99 M/96 F) were included into the cohort. Basic clinical data, parameters of glucose homeostasis, lipid concentration in plasma and conjugated diens in LDL were determined. Fatty acids were detected by capillary gas chromatography. Polymorphisms of apolipoprotein E, intestinal isoforms of fatty binding protein (Ala54Thr) and y-2 isoforms of peroxisomal activated receptor (Alal2Pro) were analyzed using combination of polymerase chain reaction methods and by the detection of polymorphisms of the restriction fragment length. Persons with had higher concentrations of CRP and conjugated diens in LDL. In all lipid classes we proved a decreased concentration of n-6 polyunsaturated fatty acids and an increase of unsaturated fatty acids. From all the acids, the only significant was the decrease of linolic concentration and the increase of and palmitoyl acids. Results showed an increase of delta 9 desaturase activity, delta 6 linolic desaturase and elongase activity. Concentration of conjugated diens in LDL inversely correlated with linolic . Clinical or laboratory parameters and homozygotic combination of polymorphism studied were not mutually related.Changes in the profile of fatty acids during the results from the elevated lipogenesis and from the higher level of oxidative stress.

Keyword: metabolic syndrome

Implication of Free Acids in Thrombin Generation and Fibrinolysis in Vascular Inflammation in Zucker Rats and Evolution with Aging.

The metabolic syndrome (MetS) and aging are associated with modifications in blood coagulation factors, vascular inflammation, and increased risk of thrombosis. Our aim was to determine concomitant changes in thrombin generation in the blood compartment and at the surface of vascular smooth muscle cells (VSMCs) and its interplay with adipokines, free acids (FFA), and metalloproteinases (MMPs) in obese Zucker rats that share features of the human MetS. Obese and age-matched lean Zucker rats were compared at 25 and 80 weeks of age. Thrombin generation was assessed by calibrated automated thrombography (CAT). Endogenous thrombin potential (ETP) was increased in obese rats independent of platelets and age. Clot half-lysis time was delayed with obesity and age. Interleukin (IL)-1β and IL-13 were increased with obesity and age respectively. Addition of exogenous fibrinogen, leptin, linoleic, or increased thrombin generation in plasma whereas adiponectin had an opposite effect. ETP was increased at the surface of VSMCs from obese rats and addition of exogenous further enhanced ETP values. Gelatinase activity was increased in aorta at both ages in obese rats and MMP-2 activity was increased in VSMCs from obese rats. Our study demonstrated in MetS an early prothrombotic phenotype of the blood compartment reinforced by procoagulant properties of dedifferentiated and inflammatory VSMCs. Mechanisms involved (1) increased fibrinogen and impaired fibrinolysis and (2) increased saturated acids responsible for additive procoagulant effects. Whether specifically targeting this hypercoagulability using direct thrombin inhibitors would improve outcome in MetS is worth investigating.

Keyword: metabolic syndrome

Free Fatty Acids: Circulating Contributors of .

induces an increased cardiovascular morbidity and mortality. Most importantly, the prevalence of in adult population is expanding. Both clinical and preclinical studies indicate that increased Free Fatty Acids (FFAs) are involved in the pathogenesis of insulin resistance and subsequent development of . The relevance of FFAs in protecting and restoring tissue function is quite vast. The search to correlate the functional deterioration of the tissues within the cardiovascular system and increased plasma concentrations of FFAs has been reported. The importance of reduction in the consumption of dietary fatty acids along with the identification of dysregulated genes responsible for persistent increased FFAs uptake and mitochondrial β-oxidation has been increasingly recognized. This review discusses the current empirical understanding of the different types of fatty acids and their metabolism and functions both in physiological and pathophysiological conditions. We also discuss in detail about the molecular and pathophysiological basis of increased FFAs, which augments Cardiovascular Disease (CVD).Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Keyword: metabolic syndrome

Analysis of Fatty Esters of Hydroxyl Fatty in Selected Plant Food.

, characterized by obesity, low-grade inflammation, insulin resistance, hyperglycemia, dyslipidemia and hypertension, is a major risk factor for cardiovascular mortality. Preclinical studies on recently discovered classes of lipids - fatty esters of hydroxy fatty acids (FAHFA) have revealed their anti-inflammatory and insulin-sensitizing potential. The FAHFA levels are significantly decreased in insulin-resistant individuals, their application exhibited anti-inflammatory effects and restoring the glucose-insulin homeostasis. The aim of our research was to analyze the overall FAHFA composition in a common diet, as only a partial FAHFA composition has been revealed so far (only the PAHSA subclass was analyzed in a few foods). A new approach to the FAHFAs analysis includes nano-LC and post-column modifier followed by negative ion mass spectrometry, in order to obtain maximum sensitivity. Analysis of different foods - oat (whole grain, coarse flakes and fine flakes), apple, clementine, lemon, strawberry, blueberry, mango, kiwi, avocado, pineapple, banana, onion, garlic, cherry tomato, carrot, parsley root, pepper and radish - exhibited wide inter-food variation in the FAHFA profiles. Sixteen analyzed FAHFAs (, oleic, palmitoleic and stearic hydroxy-esters) showed microgram to low nanogram levels (0.165\xa0ng/g - 32\xa0μg/g FW), with the highest abundancy in oat, clementine, garlic and pineapple. Stearic hydroxy stearic (SAHSA) was the most abundant FAHFA, especially in the food with antioxidative, anti-inflammatory and beneficial effects. In contrary, the PAHSA - previously proven to have the strongest antihyperglycemic and insulin-sensitizing effects, was not present in some foods (radish, avocado, mango, lemon, cherry tomato, kiwi). Our study proves the importance of overall FAHFA analysis in food (especially in a functional food), because of their potential benefits and possible future incorporation in special diets.

Keyword: metabolic syndrome

C6-C10-dicarboxylic aciduria: biochemical considerations in relation to diagnosis of beta-oxidation defects.

By means of gas chromatographic methods substantial amounts of the C6-C10-dicarboxylic acids, i.e. adipic, suberic and sebacic acids, have been found in the urine from children with unexplained attacks of lethargy and hypotonia, presumably related to episodes of fever and/or insufficient food intake. The course have once been fatal and is often characterized by severe hypoglycemia without ketonuria. Systematic gas chromatographic/mass spectrometric determinations of selected organic metabolites in the urine, together with enzymatic measurements in fibroblasts and clinical data from 4 patients of this category, have shown that the biochemical basis of this can be inborn errors of the beta-oxidation of fatty acids, localized to the medium-chain acyl-CoA dehydrogenation system. The biosynthesis of adipic, suberic and sebacic acids was studied using ketotic rats as the model, since ketosis in rats and humans is accompanied by excessive urinary excretion of adipic and suberic acids. A probable pathway for the production of the three dicarboxylic acids was found to be an initial omega-oxidation of the medium-chain C10-C14-monocarboxylic acids followed by beta-oxidation of the resulting medium-chain dicarboxylic acids. It is argued that the source of the omega-oxidizable monocarboxylic acids in ketosis most probably is the fat deposites, and it is speculated that the patients with beta-oxidation defects supplement this source with beta-oxidation intermediate medium-chain monocarboxylic acids, accumulated as a result of the defect. The ratio between the excreted amounts of adipic and sebacic in the urine from the patients with beta-oxidation defects is less than 50. This is in contrast to the ratio in urine from ketotic patients, where it is greater than 100. Adipic /sebacic ratio-measured by means of a gas chromatographic analysis-is therefore suggested as a tool in the diagnosis of dicarboxylic acidurias. Based on the clinical picture and the pattern of a series of organic acids in the urinary profile our four patients can be divided in two types of dicarboxylic aciduria. The two types have different therapeutic implications.

Keyword: metabolic syndrome

Visceral fat thickness in overweight men correlates with alterations in serum fatty composition.

We examined relationships between visceral fat amount and alterations in serum fatty composition, both of which represent critical factors in the development of .Correlations were analyzed between visceral fat thickness as measured by ultrasonography and proportions of individual fatty acids in 21 normal-weight and 24 overweight Japanese men.Significant associations were identified in overweight subjects. Visceral fat thickness displayed positive correlations to levels of and saturated fatty acids (r=0.475, P<0.05 and r=0.545, P<0.01, respectively); and negative correlations to levels of linoleic and polyunsaturated fatty acids (r=-0.513, P<0.05 and r=-0.428, P<0.05, respectively). Visceral fat thickness was also correlated with estimated desaturase activities, with positive correlations to Delta9- and Delta6-desaturase activities and negative correlations to Delta5-desaturase activity (r=0.580, P<0.01, r=0.669, P<0.01 and r=-0.559, P<0.01, respectively). No significant associations were identified in normal-weight subjects.Significant associations between visceral fat amount and alterations in serum fatty composition were identified, but only in overweight individuals.

Keyword: metabolic syndrome

Associations between the fatty content of triglyceride, visceral adipose tissue accumulation, and components of the insulin resistance .

Many factors are involved in the development of the insulin resistance , such as visceral obesity and the type of dietary fat. The main purpose of this study was to investigate the relationships between fatty content of triglyceride (TG), visceral adipose tissue (AT) accumulation, and components of the insulin resistance in a group of 97 Caucasian men with a mean age of 45.1 +/- 7.2 years (29 to 63 years). To reach these objectives, Spearman correlations, group comparisons, and stepwise multiple regression analyses were performed. The proportion of (16:0) in the TG fraction was positively associated with plasma fasting insulin (r =.25, P =.03), diastolic (r =.45, P <.001), and systolic (r =.29, P =.003) blood pressure. On the other hand, the proportion of alpha-linolenic (18:3n-3) was associated negatively with apolipoprotein (apo) B (r = -.29, P =.005) and positively with low-density lipoprotein (LDL) diameter (r =.29, P =.007), while the proportion of gamma-linolenic (18:3n-6) was associated negatively with plasma TG (r = -.33, P =.003), diastolic (r = -.29, P =.01), and systolic (r = -.35, P =.002) blood pressure and plasma fasting insulin (r = -.37, P =.0005) and positively with high-density lipoprotein (HDL)(2)-cholesterol (r =.27, P =.01) and LDL diameter (r =.25, P =.02). Stepwise multiple regression analyses were conducted to determine the contribution of visceral AT, body fat mass, and the fatty content of TG to the variance of variables studied. It was found that visceral AT contributed significantly to the variance in plasma TG (R(2) = 20.7%, P <.0001), apo B (R(2) = 9.0%, P =.007), HDL(2)-cholesterol (R(2) = 17.9%, P <.0001), LDL diameter (R(2) = 4.9%, P =.02), and area under the glucose curve (AUC-glucose) (R(2) = 8.2%, P =.006). On the other hand, body fat mass contributed significantly to the variance in fasting insulin (R(2) = 19.7%, P <.0001) and diastolic (R(2) = 6.8%, P =.007) and systolic (R(2) = 10.5%, P =.01) blood pressure. At least one fatty made a significant contribution to the variance of each variable studied. In fact, the proportion of 18:3n-6 contributed significantly to the variance in both TG (R(2) = 8.9%, P = 0.007) and HDL(2)-cholesterol (R(2) = 6.0%, P =.01). Moreover, 18:3n-3 contributed to the variance of apo B (R(2) = 7.0%, P =.02), while 18:3n-6 made the largest contribution to the variance of LDL diameter (R(2) = 7.6%, P =.02). Finally, 16:0 significantly contributed to the variance of AUC-glucose (R(2) = 11.4%, P =.0003), diastolic (R(2) = 25.2%, P <.0001), and systolic (R(2) = 6.8%, P =.002) blood pressure. In summary, results of this study suggest that the fatty content of TG is associated with many variables of the insulin resistance independently of body fat mass or visceral AT accumulation.

Keyword: metabolic syndrome

Effect of fatty acids on endothelium-dependent relaxation in the rabbit aorta.

The , Type II (non-insulin-dependent) diabetes and obesity are associated with endothelial dysfunction and increased plasma concentrations of NEFAs (non-esterified fatty acids; free fatty acids). The present study was undertaken to define the inhibitory effects of saturated NEFAs on EDR (endothelium-dependent relaxation). Experiments were performed in rings of rabbit aorta to establish (i) dose-response relationships, (ii) the effect of chain length, (iii) the effect of the presence of double bonds, (iv) reversibility and time course of inhibition, and (v) the effect on nitric oxide production. Aortic rings were incubated (1 h) with NEFA-albumin complexes derived from lauric (C(12:0)), myristic (C(14:0)), (C(16:0)), stearic (C(18:0)) and linolenic (C(18:3)) acids. EDR induced by acetylcholine (0.1-10 mumol/l) was measured after pre-contraction with noradrenaline. Inhibition of EDR was dose-dependent (0.5-2 mmol/l NEFA), and the greatest inhibition (51%) was observed with stearic (2 mmol/l). Lauric had the smallest inhibitory effect. The inhibitory effects were always reversible and were evident after 15 min of incubation. Linolenic caused a significantly lower inhibition of EDR than stearic . SOD (superoxide dismutase) restored the inhibitory effect caused by NEFAs, suggesting the involvement of ROS (reactive oxygen species) in removing nitric oxide. The nitric oxide concentration measured after exposure of the rings to acetylcholine was lower after incubation with NEFAs than with Krebs buffer alone. This finding is consistent with removal of nitric oxide by ROS. This claim was supported by the demonstration of increased concentrations of nitrated tyrosine in the rings incubated with NEFAs.

Keyword: metabolic syndrome

Lipotoxic effects of triacylglycerols in J774.2 macrophages.

Triacylglycerols (TGs) are being considered as an independent risk factor in atherosclerosis and , acting by dysregulation of the TG/high-density lipoprotein axis. Accumulation of lipids in subendothelial space attracts macrophages, leading to atherosclerotic plaque formation and increased plaque instability due to formation of foam cells and macrophage death. The aim of this study was to evaluate lipotoxic effects in macrophages caused by TG uptake.J774.2 macrophages were exposed to soybean or olive oil-based lipid emulsions as a source of TGs (1 mg/mL) in a presence or absence of lipase inhibitor paraoxon (20 microM) or to bovine serum albumin-complexed (150 microM), linoleic (600 microM), and oleic (600 microM) fatty acids.The results demonstrated accumulation of TGs, G1/S arrest, and cell death with necrotic morphologic features after exposure to TG emulsions. These effects were prevented by treatment with an antioxidant N-acetyl-cysteine (0.5 mM). Paraoxon inhibited intracellular TG degradation but did not prevent lipotoxicity and cell death. Olive oil TG triggered macrophage death in a manner similar to soybean oil. Treatment of the macrophages with free fatty , mainly with , showed a reactive oxygen species-independent cell death pathway, which was different from that of TG and was not prevented by N-acetyl-cysteine.This study shows a direct lipotoxic pathway for TG molecules in macrophages, which is not associated with degradation of TG molecule to free fatty acids. This study for the first time can explain at a cellular level how TGs as an independent risk factor aggravate atherosclerotic outcomes.

Keyword: metabolic syndrome

Downregulation of the longevity-associated protein sirtuin 1 in insulin resistance and : potential biochemical mechanisms.

Sirtuins (SIRTs) are NAD(+)-dependent deacetylases that regulate metabolism and life span. We used peripheral blood mononuclear cells (PBMCs) to determine ex vivo whether insulin resistance/ influences SIRTs. We also assessed the potential mechanisms linking alterations to SIRTs in human monocytes (THP-1) in vitro.SIRT1-SIRT7 gene and protein expression was determined in PBMCs of 54 subjects (41 with normal glucose tolerance and 13 with ). Insulin sensitivity was assessed by the minimal model analysis. Subclinical atherosclerosis was assessed by carotid intima-media thickness (IMT). In THP-1 cells exposed to high glucose or fatty acids in vitro, we explored SIRT1 expression, p53 acetylation, Jun NH(2)-terminal kinase (JNK) activation, NAD(+) levels, and nicotinamide phosphoribosyltransferase (NAMPT) expression. The effects of SIRT1 induction by resveratrol and of SIRT1 gene silencing were also assessed.In vivo, insulin resistance and were associated with low PBMC SIRT1 gene and protein expression. SIRT1 gene expression was negatively correlated with carotid IMT. In THP-1 cells, high glucose and palmitate reduced SIRT1 and NAMPT expression and reduced the levels of intracellular NAD(+) through oxidative stress. No effect was observed in cells exposed to linoleate or insulin. High glucose and palmitate increased p53 acetylation and JNK phosphorylation; these effects were abolished in siRNA SIRT1-treated cells. Glucose- and palmitate-mediated effects on NAMPT and SIRT1 were prevented by resveratrol in vitro.Insulin resistance and subclinical atherosclerosis are associated with SIRT1 downregulation in monocytes. Glucotoxicity and lypotoxicity play a relevant role in quenching SIRT1 expression.

Keyword: metabolic syndrome

[The insulin regulation of metabolism of fat acids and glucose next in the realization of biologic function of locomotion].

The becoming at the late stages of phylogeny of the biologic function of locomotion, insulin system and the earliest formed function of mitochondria make it possible to align all oxidized substrates in the following sequence: a) fatty metabolites C4 - ketone bodies; b) butyric fatty short-chained metabolites C6-C10; c) fatty with specific carrier; d) glucose. The mitochondria will begin to oxidize glucose if there will be no ketone bodies in cytosol and no remains of short-chained fatty acids and fatty . According to "the biologic subordination principle" philogenically late insulin can\'t change the functional characteristics of the phylogeny earliest mitochondria. To "force" the mitochondria starting to oxidize glucose first of all the insulin is to inhibit the biochemical reactions in all cells where releasing of polar non-etherified fatty acids and formation of their polar metabolites occurs. As in case of insulin, the same marked and prolonged hypoglycemia is induced by DL-aminocarnitine. This substance specifically inhibits both activity of carnitine-palmitoilacylaminotrsansferase and flux of acyl-KoA in mitochondria. The pronounced decrease of fatty acids content and their metabolites in matrix force mitochondria to oxidize glucose. It is possible to be validly of opinion that the same philogenically ancient principles as inhibition of activity of carnitine-palmitoilacylaminotrsansferase, decrease of formation of fatty metabolites C4 (ketone bodies), short-chained metabolites of fatty and olein mono fatty are applied in realization of philogenically late insulin effect. The first insulin effect in the hypoglycemia and biologic exotrophy reaction conditions is targeted to the regulation of fatty acids metabolism. Only second insulin effect is targeted to the glucose transformation. Therefore, there is a background to consider the diabetes mellitus primarily as a disorder of metabolism of unsaturated and mono fatty acids and only secondary and only then as a disorder of glucose metabolism. If insulin will not be able to decrease in cytosol the content of lipid substances of oxidation of insulin the mitochondria will not oxidize glucose. At that, a pathogenesis uniform of resistance to insulin is formed independently of etiologic factors. Under these conditions the mitochondria physiologically "don\'t want" to oxidize glucose a possibility exists to oxidize fatty acids and their polar metabolites.

Keyword: metabolic syndrome

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: metabolic syndrome

Distinct localisation of lipids in the ovarian follicular environment.

Obesity is associated with decreased pregnancy rates due, in part, to compromised oocyte quality. The aim of the present cross-sectional study of 84 women undergoing oocyte aspiration was to: (1) compare insulin, lipids and glucose in follicular fluid with serum; (2) determine whether increased body mass index (BMI) and waist circumference, hyperinsulinaemia, dyslipidaemia or altered follicular fluid metabolites; and (3) determine relative lipid content in oocytes to reveal any influence of these parameters on oocyte quality and IVF outcomes. Insulin, glucose, triglyceride and free fatty acids were lower in follicular fluid than blood and not strictly correlated between compartments. Insulin, glucose and triglyceride positively correlated with increasing BMI and waist circumference in blood and follicular fluid. Insulin increased in follicular fluid in association with . Free fatty composition analysis showed saturated fatty acids, particularly and stearic , to be more prevalent in follicular fluid than blood. There were no associations between follicular fluid metabolites or oocyte lipid content and clinical outcomes; however, oocyte immaturity correlated with follicular fluid glucose and fatty levels, as well as . The present study confirms that the human ovarian follicular environment surrounding the oocyte exhibits a unique metabolite profile compared with blood, with distinct localisation of lipids within follicular fluid and oocytes.

Keyword: metabolic syndrome

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance , and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: metabolic syndrome

Developmental programming: impact of prenatal testosterone excess on insulin sensitivity, adiposity, and free fatty profile in postpubertal female sheep.

Prenatal T excess causes reproductive and disruptions including insulin resistance, attributes of women with polycystic ovary . This study tested whether increases in visceral adiposity, adipocyte size, and total free fatty acids underlie the insulin resistance seen in prenatal T-treated female sheep. At approximately 16 months of age, insulin resistance and adipose tissue partitioning were determined via hyperinsulinemic euglycemic clamp and computed tomography, respectively, in control and prenatal T-treated females. Three months later, adipocyte size and free fatty composition were determined. Results revealed that at the postpubertal time points tested, insulin sensitivity was increased, visceral adiposity and adipocyte size in both the sc and the visceral compartments were reduced, and circulating was increased in prenatal T-treated females relative to controls. In parallel studies, 20-month-old prenatal T-treated females tended to have increased basal insulin to glucose ratio. Relative to earlier findings of reduced insulin sensitivity of prenatal T-treated females during early life and adulthood, these findings of increased insulin sensitivity and reduced adiposity postpubertally are suggestive of a period of developmental adaptation. The disruption observed in free fatty metabolism a few months later correspond to a time point when the insulin sensitivity indices of prenatal T-treated animals appear to shift toward insulin resistance. In summary, current findings of improved insulin sensitivity and reduced visceral adiposity in postpubertal prenatal T-treated sheep relative to our earlier findings of reduced insulin sensitivity during early postnatal life and adulthood are indicative of a period of developmental adaptation.

Keyword: metabolic syndrome

MicroRNA-194 inhibition improves dietary-induced non-alcoholic disease in mice through targeting on FXR.

Non-alcoholic disease (NAFLD) affects obesity-associated metabolic syndrome, which exhibits hepatic steatosis, insulin insensitivity and glucose intolerance. Previous studies indicated that hepatic microRNAs (miRs) play critical roles in the development of NAFLD. In this study, we aim to explore the pathophysiological role of miR-194 in obesity-mediated metabolic dysfunction. Our findings show that the high fat diet or treatment significantly increase hepatic miR-194 levels in vivo and in vitro. Silence of miR-194 protects -induced inflammatory response in cultured hepatocytes, and attenuates structural disorders, lipid deposits and inflammatory response in . MiR-194 inhibitor also improves glucose and insulin intolerance in obese mice. Through dual luciferase assay, we demonstrate that miR-194 directly binds to FXR/Nr1h4 3\'-UTR, and inhibits gene expression of FXR/Nr1h4. Furthermore, overexpression of miR-194 downregulates FXR/Nr1h4 in cultured hepatocytes, but miR-194 inhibitor reversely increases FXR/Nr1h4 expression in obese mouse tissues. On the contrast, silence of FXR/Nr1h4 abolishes the hepatic benefits in obese mice treated with miR-194 inhibitor. Present study provides a novel finding that suppression of miR-194 attenuates dietary-induced NAFLD via upregulation of FXR/Nr1h4. The findings suggest miR-194/FXR are potential diagnostic markers and therapeutic targets for NAFLD.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Fatty Composition of Plasma Phosphatidylcholine Determines Body Fat Parameters in Subjects with -Related Traits.

This study examines the associations of fatty acids (FAs) in plasma phosphatidylcholine (PC) with the anthropometrical and biochemical characteristic of patients with (MetS)-related traits.We analyzed the FA profiles of PC in 300 persons with MetS-related traits (152\u2009M/148F, mean age 46.9\u2009±\u20099.0 years) and in 70 healthy controls of the same age using a balanced men/women ratio and gas-liquid chromatography. Multivariate linear regression analysis was performed to determine the coefficients of determination (R) using FA proportions of the mentioned proband characteristics.The FA composition of PC in patients with MetS traits was only associated with waist circumference (R\u2009=\u20090.27), waist-to-hip ratio (WHR; R\u2009=\u20090.41), body fat percentage (R\u2009=\u20090.62), and fat mass (R\u2009=\u20090.29). Positive associations were found for dihomo-γ-linolenic (DGLA), , stearic (SA), α-linolenic (ALA), and eicosapentaenoic acids, whereas negative associations were found for linoleic (LA), oleic, and docosapentaenoic acids. Palmitoleic (POA) was positively associated with waist circumference but negatively with fat percentage. In controls, significant associations were found for waist circumference (R\u2009=\u20090.51), WHR (R\u2009=\u20090.53), body fat percentage (R\u2009=\u20090.60), and fat mass (R\u2009=\u20090.34). DGLA and saturated FA (SFA) were positively associated, whereas docosahexaenoic, adrenic, and cis-vaccenic acids were negatively associated. The study group differed from controls as follows: lower concentrations of LA and total n-6 FA, higher indices of delta-9-desaturase and delta-6 desaturase activity and higher proportions of POA, SA, ALA, DGLA, and SFA.We found significant associations (R >0.25) of FA in plasma PC with adiposity in middle-aged persons with MetS-related traits, but not with indices.

Keyword: metabolic syndrome

Folic supplementation during high-fat diet feeding restores AMPK activation via an AMP-LKB1-dependent mechanism.

AMPK is an endogenous energy sensor that regulates lipid and carbohydrate metabolism. Nonalcoholic fatty liver disease (NAFLD) is regarded as a hepatic manifestation of with impaired lipid and glucose metabolism and increased oxidative stress. Our recent study showed that folic supplementation attenuated hepatic oxidative stress and lipid accumulation in high-fat diet-fed mice. The aim of the present study was to investigate the effect of folic on hepatic AMPK during high-fat diet feeding and the mechanisms involved. Male C57BL/6J mice were fed a control diet (10% kcal fat), a high-fat diet (60% kcal fat), or a high-fat diet supplemented with folic (26 mg/kg diet) for 5 wk. Mice fed a high-fat diet exhibited hyperglycemia, hepatic cholesterol accumulation, and reduced hepatic AMPK phosphorylation. Folic supplementation restored AMPK phosphorylation (activation) and reduced blood glucose and hepatic cholesterol levels. Activation of AMPK by folic was mediated through an elevation of its allosteric activator AMP and activation of its upstream kinase, namely, liver kinase B1 (LKB1) in the liver. Consistent with in vivo findings, 5-methyltetrahydrofolate (bioactive form of folate) restored phosphorylation (activation) of both AMPK and LKB1 in -treated HepG2 cells. Activation of AMPK by folic might be responsible for AMPK-dependent phosphorylation of HMG-CoA reductase, leading to reduced hepatic cholesterol synthesis during high-fat diet feeding. These results suggest that folic supplementation may improve cholesterol and glucose metabolism by restoration of AMPK activation in the liver.Copyright © 2015 the American Physiological Society.

Keyword: metabolic syndrome

Perivascular adipose tissue dysfunction aggravates adventitial remodeling in obese mini pigs via NLRP3 inflammasome/IL-1 signaling pathway.

Perivascular adipose tissue (PVAT), a special type of adipose tissue, closely surrounds vascular adventitia and produces numerous bioactive substances to maintain vascular homeostasis. PVAT dysfunction has a crucial role in regulating vascular remodeling, but the exact mechanisms remain unclear. In this study, we investigated whether and how obesity-induced PVAT dysfunction affected adventitia remodeling in early vascular injury stages. Mini pigs were fed a high sugar and fat diet for 6 months to induce and obesity. In the mini pigs, left carotid vascular injury was then generated using balloon dilation. Compared with normal mini pigs, obese mini pigs displayed significantly enhanced vascular injury-induced adventitial responses, evidenced by adventitia fibroblast (AF) proliferation and differentiation, and adventitia fibrosis, as well as exacerbated PVAT dysfunction characterized by increased accumulation of resident macrophages, particularly the M1 pro-inflammatory phenotype, increased expression of leptin and decreased expression of adiponectin, and production of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Primary AFs cultured in PVAT-conditioned medium from obese mini pigs also showed significantly increased proliferation and differentiation. We further revealed that activated nod-like receptor protein 3 (NLRP3) inflammasome and its downstream products, i.e., IL-1 family members such as IL-1β and IL-18 were upregulated in the PVAT of obese mini pigs; PVAT dysfunction was also demonstrated in preadipocytes treated with . Finally, we showed that pretreatment with IL-1 receptor (IL-1R) antagonist or IL-1R knockdown blocked AF proliferation and differentiation in AFs cultured in PVAT-conditioned medium. These results demonstrate that obesity-induced PVAT dysfunction aggravates adventitial remodeling after early vascular injury with elevated AF proliferation and differentiation via activating the NLRP3/IL-1 signaling pathway.

Keyword: metabolic syndrome

exacerbates inflammation and bone loss in periodontitis.

Clinical studies have shown that (MetS) is associated with increased risk of developing periodontitis. However, the underlying mechanisms remain largely unknown. Since it is known that lipopolysaccharide (LPS)-activated toll-like receptor 4 signaling pathways play a crucial role in periodontitis, we hypothesized that MetS enhances LPS-induced periodontal inflammation and alveolar bone loss. In this study, we induced MetS in C57BL/6 mice by feeding them high-fat diet (HFD), and we induced periodontitis by periodontal injection of Aggregatibacter actinomycetemcomitans LPS. We found that mice fed a HFD had significantly increased body weight, plasma lipids, insulin, and insulin resistance when compared with mice fed regular chow, indicating that the mice developed MetS. We also found that a HFD markedly increased LPS-induced alveolar bone loss, osteoclastogenesis, and inflammatory infiltration. Analysis of gene expression in periodontal tissue revealed that HFD and LPS injection cooperatively stimulated expression of cytokines that are known to be involved in periodontal tissue inflammation and osteoclastogenesis-such as interleukin 6, monocyte-chemotactic protein 1, receptor activator of nuclear factor kappa-B ligand, and macrophage colony-stimulating factor. To further understand the potential mechanisms involved in MetS-boosted tissue inflammation, our in vitro studies showed that -the most abundant saturated fatty (SFA) and the major SFA in the HFD used in our animal study-potently enhanced LPS-induced proinflammatory gene expression in macrophages. In sum, this study demonstrated that MetS was associated with increased periodontal inflammation and alveolar bone loss in an LPS-induced periodontitis animal model. This study also suggests that SFA may play an important role in MetS-associated periodontitis by enhancing LPS-induced expression of inflammatory cytokines in macrophages.© International & American Associations for Dental Research 2014.

Keyword: metabolic syndrome

omega-123I-hexadecanoic probe of cardiomyopathy.

The utility of omega-123I-hexadecanoic myocardial scintigraphy as a probe of cardiomyopathies was investigated. Sixteen patients with a variety of cardiomyopathies and myopathies that involve cardiac muscle and ten volunteers were imaged in the postabsorptive state in a 40 degrees LAO projection after a standard dose of omega-123I-hexadecanoic . An elimination T1/2 was calculated from the left ventricular myocardial time-activity curve. An uptake index, corrected for chest wall attenuation, was also computed in 7 of 10 volunteers and 8 of 16 patients. Of the 16 patients, only 2 had distinctly abnormal omega-123I-hexadecanoic myocardial tracer kinetics. The first patient had a disorder of which carnitine deficiency was one component. The second patient had endocardial fibroelastosis, a process which has been linked to disorders which deprive the myocardium of oxygen and energy. Therefore, the cardiomyopathy may have been caused by some abnormality of cardiac metabolism other than carnitine deficiency. Although of limited utility in the overall cardiomyopathic population, omega-123I-hexadecanoic myocardial scintigraphy should be further investigated as a screening test for carnitine deficiency and related abnormalities in patients at risk.

Keyword: metabolic syndrome

[Prevention of atherosclerosis. Excess of in food--a cause of hypercholesterolemia, inflammatory , insulin resistance in myocytes, and apoptosis].

Unity of the pathogenesis of atherosclerosis, type 2 diabetes mellitus, and gives rise to impaired biological function of adaptation, altered biological function of exotrophy (external feeding) and endoecology ("purity" of the intercellular medium). Biological reactions of inflammation and hydrodynamic pressure, or blood pressure, are in vivo activated to compensate for intercellular debris accumulation by endogenous phlogogens--ligand-free low density lipoproteins (LDL). The biological reactions jointly remove LDL from blood to the intima of elastic type arteries, to interstitial tissue for the local pool of the intravascular medium. The causes of formation of aphysiological LDLs are a preponderance of palmitate-oleate-palmitate triglycerides in the latter and impaired hydrolysis upon exposure to post-heparin lipase to give rise to small, dense LDLs; intimal macrophages utilize the debris only partially and develop atheromatosis from polyenic fatty acids (FA) etherified by cholesterol alcohol. Excess of saturated fatty (sFA) is responsible for the lowered permeability of the plasma membrane, cell death via the mechanism similar to apoptosis. Aphysiological protein palmitoylation (covalent interaction with sFA) increased the debris accumulation of the intercellular medium and the activity of both biological reactions. Elevated plasma sFA and its enhanced passive absorption in the form of unetherified FA, as well as high C-reactive protein levels are a cause of insulin resistance. The only way to prevent atherosclerosis in the population is to normalize the biological function of exotrophy when the energy value ratio of FA, proteins and carbohydrates is 1:1:1 and that of sFA, monoenic, and polyenic FA is also 1:1:1. The lower amount of sFA and the higher concentration of essential polyenic FA, the lower blood levels of cholesterol alcohol and triglycerides are. At the same time, simultaneously activations and the biological function of locomotion are a level of physical activity.

Keyword: metabolic syndrome

Increased RhoA/Rho-Kinase Activity and Markers of Endothelial Dysfunction in Young Adult Subjects with .

, a chronic condition associated with higher risk of cardiovascular diseases, is increasingly prevalent in young adults. Dyslipidemia, proinflammatory cytokines, endothelial dysfunction signs, and RhoA/Rho-kinase (ROCK) activation are considered risk factors of cardiovascular diseases. The occurrence of these factors in young patients with but without type 2 diabetes or hypertension has not been fully studied. The objective of this study was to evaluate young subjects with enlarged waist circumference and dyslipidemia but without type 2 diabetes or hypertension,for markers associated with a higher risk of cardiovascular diseases.Thirty-two male patients aged 31\u2009±\u20091.3 years diagnosed with according to the National Cholesterol Education Program Adult Treatment Panel III guide for enlarged waist circumference, elevated triglycerides, and low HDL levels, but with blood pressure and fasting glucose within normal ranges, were evaluated for RhoA/ROCK activity in leukocytes, serum fatty methyl esters profile, proinflammatory cytokines, and oxidative stress markers in addition to thrombin generation and biochemical analysis. Age- and gender-matched healthy subjects were equivalently evaluated.Patients showed higher RhoA/ROCK activity, elevated levels of interleukin-6, soluble CD40L, monocyte chemoattractant protein, and high-sensitivity C-reactive protein (P\u2009<\u20090.001) as well as parameters of endogenous thrombin generation potential (P\u2009<\u20090.05) compared with healthy subjects. Increased thiobarbituric reactive substances, advanced oxidation protein product, and insulin levels and low nitric oxide biodisponibility (P\u2009<\u20090.001) were also found in patients as compared with controls. was one of the saturated fatty acids found to be significantly elevated in patients compared with control subjects (P\u2009=\u20090.0087).Increased markers of cardiovascular risk are already present in young adults with but without type 2 diabetes or hypertension.

Keyword: metabolic syndrome

[Methylglyoxal--test for biological dysfunctions of homeostasis and endoecology, low cytosolic glucose level, and gluconeogenesis from fatty acids].

If a lot of carbohydrates cannot be in vivo stored as glycogen, the synthesis of fatty (FA) from glucose and its adipocyte deposition as triglycerides are under way in phylogenesis. With impaired biological function of exotrophy (fasting, early postnatality, hibernation), the cells perform a reverse process--the synthesis of glucose from FA. Physiologically, the substrate of gluconeogenesis is acetyl-CoA that is converted by the malate --> 9 piruvate --> glucose pathway in the glyoxalate cycle. Under the pathological conditions of hypoxia and energy deficiency, gluconeogenesis occurs without ATP consumption via the methylglyoxalate pathway (MGP) while using as a substrate of ketone bodies: butyric (butyrate) --> beta-hydroxybutyrate --> acetoacetate --> acetone --> acetol --> methylglyoxal (MG) --> S-D-lactolglutathione --> D-lactate --> piruvate --> D-lactate. Under physiological conditions, this pathway of gluconeogenesis does not work. The authors hold that gene expression and gluconeogenesis occur via the MGP when glucose levels are low in the cell cytosol (glycopenia) and FA cannot be oxidized in the mitochondria. Cytosol, intercellular medium, plasma show elevated levels of MG and D-lactate, to which it converts under the action of glyoxalases I and II. Glycopenia develops in fasting, diabetes mellitus, , renal failure, phenofibrate therapy, impaired function of exotrophy--excessive dietary intake of saturated and trans fatty acids. The chemical interaction of MG with amino residues of lysine and arginine leads to protein denaturation during carbonylation--glycosylation and impaired biological function of endoecology. The determination of plasma MG and D-lactate may be a test for glycopenia, compensatory activation of gluconeogenesis from FA or for the evaluation of endogenous intoxication.

Keyword: metabolic syndrome

Ruscogenin ameliorates experimental nonalcoholic steatohepatitis via suppressing lipogenesis and inflammatory pathway.

The aim of the study was to investigate the protective effects of ruscogenin, a major steroid sapogenin in Ophiopogon japonicus, on experimental models of nonalcoholic steatohepatitis. HepG2 cells were exposed to 300\u2009μmol/l (PA) for 24\u2009h with the preincubation of ruscogenin for another 24\u2009h. Ruscogenin (10.0\u2009μmol/l) had inhibitory effects on PA-induced triglyceride accumulation and inflammatory markers in HepG2 cells. Male golden hamsters were randomly divided into five groups fed a normal diet, a high-fat diet (HFD), or a HFD supplemented with ruscogenin (0.3, 1.0, or 3.0\u2009mg/kg/day) by gavage once daily for 8 weeks. Ruscogenin alleviated dyslipidemia, liver steatosis, and necroinflammation and reversed plasma markers of in HFD-fed hamsters. Hepatic mRNA levels involved in fatty oxidation were increased in ruscogenin-treated HFD-fed hamsters. Conversely, ruscogenin decreased expression of genes involved in hepatic lipogenesis. Gene expression of inflammatory cytokines, chemoattractive mediator, nuclear transcription factor-(NF-) κB, and α-smooth muscle actin were increased in the HFD group, which were attenuated by ruscogenin. Ruscogenin may attenuate HFD-induced steatohepatitis through downregulation of NF-κB-mediated inflammatory responses, reducing hepatic lipogenic gene expression, and upregulating proteins in β-oxidation pathway.

Keyword: metabolic syndrome

[Pharmacologic influencing of fetal phospholipide synthesis. Part II: Carnitin - a new way of dyspnea prophylaxis? (author\'s transl)].

Carnitine is a body-owned Betaine which is wide-spread in living nature in the form of a physiologic metabolit. On account of its central function, Carnitine has various effects in the intermediary metabolism. Toxicity of Carnitine is very low. DL-Carnitine hydrochloride (30 mg/kg body weight) was injected into 25 gravid Wistar rats and 24 gravid rabbits, 4 days before the Cesarean section. Compared to the group of controls, the group treated with Bromhexine metabolit VIII, as well as to the group treated with Betamethason, significantly higher values with regard to the content of total phospholipids, lecithine and could be found in the fetal rat lungs. Effects of Carnitine on the phospholipid metabolism are discussed.

Keyword: metabolic syndrome

Intravenous Mycobacterium Bovis Bacillus Calmette-Guérin Ameliorates Nonalcoholic Fatty Liver Disease in Obese, Diabetic ob/ob Mice.

Inflammation and immune response profoundly influence and fatty metabolism. To analyze influence of systemic inflammatory response to , we inoculated an attenuated vaccine strain of Mycobacterium bovis Bacillus Calmette-Guérin (BCG) into leptin-deficient ob/ob mice. BCG administration significantly decreased epididymal white adipose tissue weight, serum insulin levels, and a homeostasis model assessment of insulin resistance. Serum high molecular weight (HMW) adiponectin level and HMW/total adiponectin ratio of the BCG treated mice were significantly higher than those of control mice. Hepatic triglyceride accumulation and macrovesicular steatosis were markedly alleviated, and the enzymatic activities and mRNA levels of lipogenic-related genes in liver were significantly decreased in the BCG injected mice. We also exposed human hepatocellular carcinoma HepG2 cells to high levels of palmitate, which enhanced endoplasmic reticulum stress-related gene expression and impaired insulin-stimulated Akt phosphorylation (Ser473). BCG treatment ameliorated both of these detrimental events. The present study therefore suggested that BCG administration suppressed development of nonalcoholic fatty liver disease, at least partly, by alleviating fatty -induced insulin resistance in the liver.

Keyword: metabolic syndrome

Glucose and fatty metabolism in McA-RH7777 hepatoma cells vs. rat primary hepatocytes: responsiveness to nutrient availability.

The overabundance of dietary fats and simple carbohydrates contributes significantly to obesity and disorders associated with obesity. The liver balances glucose and lipid distribution, and disruption of this balance plays a key role in these syndromes. We investigated (1) how hepatocytes balance glucose and fatty metabolism when one or both nutrients are supplied in abundance and (2) whether rat hepatoma cells (McA-RH7777) reflect nutrient partitioning in a similar manner as compared with primary hepatocytes. Increasing media palmitate concentration increased fatty uptake, triglyceride synthesis and beta-oxidation. However, hepatoma cells had a 2-fold higher fatty uptake and a 2-fold lower fatty oxidation as compared with primary hepatocytes. McA-RH7777 cells did not synthesize significant amounts of glycogen and preferentially metabolized the glucose into lipids or into oxidation. In primary hepatocytes, the glucose was mostly spared from oxidation and instead partitioned into both de novo glycogen and lipid synthesis. Overall, lipid production was rapidly induced in response to either glucose or fatty excess and this may be one of the earliest indicators of development associated with nutrient excess.

Keyword: metabolic syndrome

MSP is a negative regulator of inflammation and lipogenesis in ex vivo models of non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (NASH), a disorder consisting of steatosis and inflammation, is considered the hepatic equivalent of and can result in irreversible liver damage. Macrophage-stimulating protein (MSP) is a hepatokine that potentially has a beneficial role in hepatic lipid and glucose metabolism via the activation of AMP-activated protein kinase (AMPK). In the current study, we investigated the regulatory role of MSP in the development of inflammation and lipid metabolism in various NASH models, both in vitro and ex vivo. We observed that MSP treatment activated the AMPK signaling pathway and inhibited lipopolysaccharide (LPS)- and (PA)-induced gene expression of pro-inflammatory cytokines in primary mouse hepatocytes. In addition, MSP treatment resulted in a significant reduction in PA-induced lipid accumulation and inhibited the gene expression of key lipogenic enzymes in HepG2 cells. Upon short hairpin RNA-induced knockdown of RON (the membrane-bound receptor for MSP), the anti-inflammatory and anti-lipogenic effects of MSP were markedly ablated. Finally, to mimic NASH ex vivo, we challenged bone marrow-derived macrophages with oxidized low-density lipoprotein (oxLDL) in combination with LPS. OxLDL+LPS exposure led to a marked inhibition of AMPK activity and a robust increase in inflammation. MSP treatment significantly reversed these effects by restoring AMPK activity and by suppressing pro-inflammatory cytokine gene expression and secretion under this condition. Taken together, these data suggest that MSP is an effective inhibitor of inflammation and lipid accumulation in the stressed liver, thereby indicating that MSP has a key regulatory role in NASH.

Keyword: metabolic syndrome

Comparison of Fatty Profiles in a Group of Female Patients with Chronic Kidney Diseases (CKD) and (MetS)⁻Similar Trends of Changes, Different Pathophysiology.

Fatty (FA) profiles in the plasma of patients with and chronic kidney disease (CKD) seem to be identical despite their different etiology (dietary mistakes vs. cachexia). The aim of this study was to compare both profiles and to highlight the differences that could influence the improvement of the treatment of patients in both groups. The study involved 73 women, including 24 patients with chronic kidney disease treated with haemodialysis, 19 patients with (MetS), and 30 healthy women in the control group. A total of 35 fatty acids and derivatives were identified and quantified by gas chromatography. Intensified elongation processes from C10:0 to C16:0 were noted in both groups (more intense in MetS), as well as an increased synthesis of arachidonic (C20:4n6), which was more intense in CKD. Significant correlations of oleic (C18:1n9), gamma linoleic (C18:3n6), and docosatetraenoate (C22:4n6) with parameters of CKD patients were observed. In the MetS group, auxiliary pathways of oleic were activated, which simultaneously inhibited the synthesis of eicosapentanoic (EPA) and docosahexaenoic (DHA) from alpha lipoic (ALA). On the other hand, in the group of female patients with CKD, the synthesis of EPA and DHA was intensified. Activation of the synthesis of oleic (C18: 1n9 ct) and trans-vaccinic (C18:1) is a protective mechanism in kidney diseases and especially in MetS due to the increased concentration of saturated fatty (SFA) in plasma. The cause of the increased amount of all FAs in plasma in the CKD group, especially in the case of (C16:0) and derivatives stearic (C18:0) acids, may be the decomposition of adipose tissue and the progressing devastation of the organism, whereas, in the MetS group, dietary intake seems to be the main reason for the increase in SFA. Moreover, in MetS, auxiliary pathways are activated for oleic , which cause the simultaneous inhibition of EPA and DHA synthesis from ALA, whereas, in the CKD group, we observe an increased synthesis of EPA and DHA. The higher increase of nervonic (C24:1) in CKD suggests a higher degree of demyelination and loss of axons.

Keyword: metabolic syndrome

Palmitate increases the susceptibility of cells to drug-induced toxicity: an in vitro method to identify drugs with potential contraindications in patients with disease.

Fatty acids are an important source of energy. Excessive energy intake results in elevated levels of free fatty acids that are thought to be the pathogenic factors causing disorders such as dyslipidemia, obesity, insulin resistance, diabetes, and fatty liver. Underlying disorders have been suggested to be a predisposing factor for drug-induced liver injury. The steadily expanding population with disease may pose a higher risk for drug-induced toxicity. In order to understand the interaction of free fatty acids and drug-induced toxicity at the cellular level, we explored whether the saturated free fatty palmitate could modulate drug-induced cytotoxicity in HepG2 cells. A number of drugs known to induce hepatotoxicity in humans were selected to test this hypothesis. Drugs without reported hepatotoxicity were also tested to evaluate the specificity of the palmitate-induced effects. We demonstrate that palmitate, at sublethal concentrations, was able to potentiate the cytotoxicity and/or apoptosis induced by some but not all drugs tested. The palmitate and drug coincubation potentiated toxicity, which when combined with the plasma maximum concentration (C(max)), allowed us to identify idiosyncratic toxic drugs that were not flagged in previously deployed cytotoxicity assays. Our data suggest that treatment of cells with palmitate improves the sensitivity to detect compounds with risk of inducing idiosyncratic liver toxicity. Furthermore, this assay may be used to identify compounds that have higher safety risks in a population with .

Keyword: metabolic syndrome

Metformin reduces intracellular reactive oxygen species levels by upregulating expression of the antioxidant thioredoxin via the AMPK-FOXO3 pathway.

Oxidative stress induced by free fatty acids plays a critical role in the pathogenesis of endothelial dysfunction and atherosclerosis in patients with . Reducing oxidative stress in these patients may prevent cardiovascular complications. The antidiabetic agent metformin has been reported to directly protect the cardiovascular system. In this study, we examined the effect of metformin on the intracellular levels of reactive oxygen species (ROS) induced by (PA) in human aortic endothelial cells and studied the molecular mechanisms involved.We observed that metformin significantly reduced intracellular ROS levels induced by PA. Additionally, metformin increased the expression of the antioxidant thioredoxin (Trx), which mediated metformin\'s effects on ROS reduction. Metformin increased Trx expression through the AMP-activated protein kinase (AMPK) pathway. Metformin-regulated Trx at the transcriptional level and forkhead transcription factor 3 (FOXO3) was involved in this process.These results suggest that metformin reduces ROS levels by inducing Trx expression through activation of the AMPK-FOXO3 pathway.Copyright (c) 2010 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Activation of the AMPK-FOXO3 pathway reduces fatty -induced increase in intracellular reactive oxygen species by upregulating thioredoxin.

Oxidative stress induced by free fatty acids contributes to the development of cardiovascular diseases in patients with . Reducing oxidative stress may attenuate these pathogenic processes. Activation of AMP-activated protein kinase (AMPK) has been reported to reduce intracellular reactive oxygen species (ROS) levels. The thioredoxin (Trx) system is a major antioxidant system. In this study, we investigated the mechanisms involved in the AMPK-mediated regulation of Trx expression and the reduction of intracellular ROS levels.We observed that activation of AMPK by 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) significantly reduced ROS levels induced by in human aortic endothelial cells. Activation of AMPK increased expression of the antioxidant Trx, which mediated the ROS reduction. RT-PCR showed that AMPK regulated Trx at the transcriptional level.Forkhead transcription factor 3 (FOXO3) was identified as the target transcription factor involved in the upregulation of Trx expression. FOXO3 bound to the Trx promoter, recruited the histone acetylase p300 to the Trx promoter, and formed a transcription activator complex, which was enhanced by AICAR treatment. AMPK activated FOXO3 by promoting its nuclear translocation. We further showed that AICAR injection increased the expression of Trx and decreased ROS production in the aortic wall of ApoE-/- mice fed a high-fat diet.These results suggest that activation of the AMPK-FOXO3 pathway reduces ROS levels by inducing Trx expression. Thus, the AMPK-FOXO3-Trx axis may be an important defense mechanism against excessive ROS production induced by stress and could be a therapeutic target in treating cardiovascular diseases in .

Keyword: metabolic syndrome

Inhibition of fatty beta oxidation by influenza B virus and salicylic in mice: implications for Reye\'s .

Injection of concentrated influenza B/Lee/40 virus into 4-week-old Balb C mice resulted in 60% inhibition of 14C-palmitate oxidation in isolated hepatic mitochondria. Oral feeding of carnitine to infected mice prevented the inhibition of fatty oxidation. High concentrations of salicylic given orally also inhibited 14C-palmitate oxidation. Serum free fatty concentrations of infected mice and of those fed salicylic were significantly higher than in control mice. A combination of low-dose virus and low-dose salicylic inhibited palmitate oxidation, suggesting an additive effect on the derangement when the two agents were present simultaneously.

Keyword: metabolic syndrome

Time-dependent effects of fatty acids on skeletal muscle metabolism.

Increased plasma levels of free fatty acids (FFA) occur in states of insulin resistance such as type 2 diabetes mellitus, obesity, and . These high levels of plasma FFA seem to play an important role for the development of insulin resistance but the mechanisms involved are not known. We demonstrated that acute exposure to FFA (1 h) in rat incubated skeletal muscle leads to an increase in the insulin-stimulated glycogen synthesis and glucose oxidation. In conditions of prolonged exposure to FFA, however, the insulin-stimulated glucose uptake and metabolism is impaired in skeletal muscle. In this review, we discuss the differences between the effects of acute and prolonged exposure to FFA on skeletal muscle glucose metabolism and the possible mechanisms involved in the FFA-induced insulin resistance.

Keyword: metabolic syndrome

Acne vulgaris: The metabolic syndrome of the pilosebaceous follicle.

Acne vulgaris is an epidemic inflammatory disease of the human sebaceous follicle and represents the most common skin disease affecting about 85% of adolescents in Westernized populations. Acne vulgaris is primarily a disease of wealthy countries and exhibits higher prevalence rates in developed compared with developing countries. No acne has been found in non-Westernized populations still living under Paleolithic dietary conditions constraining hyperglycemic carbohydrates, milk, and dairy products. The high prevalence rates of adolescent acne cannot be explained by the predominance of genetic factors but by the influence of a Western diet that overstimulates the key conductor of metabolism, the nutrient- and growth factor-sensitive kinase mTORC1. Increased mTORC1 activity has been detected in lesional skin and sebaceous glands of acne patients compared with acne-free controls. Increased mTORC1 signaling is a characteristic feature of insulin resistance, obesity, type 2 diabetes mellitus, cancer, and neurodegenerative diseases. Acne vulgaris is a family member of mTORC1-driven diseases of civilization and represents the MetS of the sebaceous follicle.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Individual fatty acids in erythrocyte membranes are associated with several features of the in obese children.

Obesity leads to the clustering of cardiovascular (CV) risk factors and the (MetS) also in children and is often accompanied by non-alcoholic fatty liver disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 fatty acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting insulin (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting insulin (r\u2009=\u20090.287). Fatty Liver Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

Keyword: metabolic syndrome

An inhibitor of phospholipase A2 group IIA modulates adipocyte signaling and protects against diet-induced in rats.

Obesity, type 2 diabetes, and cardiovascular disease correlate with infiltration to adipose tissue of different immune cells, with uncertain influences on metabolism. Rats were fed a diet high in carbohydrates and saturated fats to develop diet-induced obesity over 16 weeks. This nutritional overload caused overexpression and secretion of phospholipase A(2) group IIA (pla2g2a) from immune cells in adipose tissue rather than adipocytes, whereas expression of adipose-specific phospholipase A(2) (pla2g16) was unchanged. These immune cells produce prostaglandin E(2) (PGE(2)), which influences adipocyte signaling. We found that a selective inhibitor of human pla2g2a (5-(4-benzyloxyphenyl)-(4S)-(phenyl-heptanoylamino)-pentanoic [KH064]) attenuated secretion of PGE(2) from human immune cells stimulated with the fatty , , or with lipopolysaccharide. Oral administration of KH064 (5 mg/kg/day) to rats fed the high-carbohydrate, high-fat diet prevented the overexpression of pla2g2a and the increased macrophage infiltration and elevated PGE(2) concentrations in adipose tissue. The treatment also attenuated visceral adiposity and reversed most characteristics of , producing marked improvements in insulin sensitivity, glucose intolerance, and cardiovascular abnormalities. We suggest that pla2g2a may have a causal relationship with chronic adiposity and and that its inhibition in vivo may be a valuable new approach to treat obesity, type 2 diabetes, and dysfunction in humans.

Keyword: metabolic syndrome

modulation predicts heart failure tests performance.

The changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Pathway Analysis (MetPA) used predictors to identify the most relevant pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the underpinning of CPET and HFBio.

Keyword: metabolic syndrome

[THE OPTIMIZATION OF NUTRITION FUNCTION UNDER OF RESISTANCE TO INSULIN, DISORDER OF FATTY ACIDS\' METABOLISM AND ABSORPTION OF GLUCOSE BY CELLS (A LECTURE)].

The phylogenetic processes continue to proceed in Homo Sapiens. At the very early stages ofphylogenesis, the ancient Archaea that formed mitochondria under symbiotic interaction with later bacterial cells conjointly formed yet another system. In this system, there are no cells\' absorption of glucose if it is possible to absorb fatty acids from intercellular medium in the form of unesterfied fatty acids or ketonic bodies--metabolites of fatty acids. This is caused by objectively existed conditions and subsequent availability of substrates at the stages ofphylogenesis: acetate, ketonic bodies, fatty acids and only later glucose. The phylogenetically late insulin used after billions years the same dependencies at formation of regulation ofmetabolism offatty acids and cells\' absorption of glucose. In order that ofresistance ceased to exist as afoundation of pandemic Homo Sapiens has to understand the following. After successful function ofArchaea+bacterial cells and considered by biology action of insulin for the third time in phylogenesis and using biological function of intelligence the content ofphylogenetically earlier saturated fatty infood can\'t to exceed possibilities of phylogenetically late lipoproteins to transfer it in intercellular medium and blood and cells to absorb it. It is supposed that at early stages of phylogenesis biological function of intelligence is primarily formed to bring into line "unconformities" of regulation of metabolism against the background of seeming relative biological "perfection". These unconformities were subsequently and separately formed at the level of cells in paracrin regulated cenosises of cells and organs and at the level of organism. The prevention of resistance to insulin basically requires biological function of intelligence, principle of self-restraint, bringing into line multiple desires of Homo Sapiens with much less extensive biological possibilities. The "unconformities" of regulation of metabolism in vivo are etiological factors of all pandemics including atherosclerosis, arterial hypertension, obesity and Tertiannondatum.

Keyword: metabolic syndrome

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) , the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) , 4) obesity, 5) insulin resistance , 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: metabolic syndrome

The Haematococcus pluvialis extract enriched by bioaccumulation process with Mg(II) ions improves insulin resistance in equine adipose-derived stromal cells (EqASCs).

Insulin resistance (IR) is one of the characteristic features of equine (EMS). Presently, the only therapies of choice are caloric restrictions combined with mineral supplementation, which might improve insulin sensitivity. In this study we investigated the effect of Haematococcus pluvialis algae water extract enriched in bioaccumulation process in magnesium ions (Hp_Mg(II)) on equine adipose derived mesenchymal stromal stem cells, in which insulin resistance was induced by (IR-EqASCs). For this purpose, chemical characterization of H. pluvialis was performed with special emphasis on the analysis of minerals composition, total phenolic and carotenoids contents, as well as scavenging activity. To examine the influence of H. pluvialis extract on IR-EqASCs, various methods of molecular biology and microscopic observations (i.e., immunofluorescence staining, SEM, gene expression by RT-qPCR, proliferative and cells activity analysis) were applied to investigate in vitro viability, oxidative stress markers and apoptosis-related factor accumulation, along with insulin resistance-related genes expression. Obtained results show, that Hp_Mg(II) significantly improves proliferative and activity of IR-EqASCs, shortens their population doubling time, improves their clonogenic potential and reduces expression of apoptosis related genes. Moreover, anti-oxidative effect of extract was presented.Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: metabolic syndrome

Palmitate aggravates proteinuria-induced cell death and inflammation via CD36-inflammasome axis in the proximal tubular cells of obese mice.

High levels of serum free fatty acids (FFAs) and proteinuria have been implicated in the pathogenesis of obesity-related nephropathy. CD36, a class B scavenger receptor, is highly expressed in the renal proximal tubules and mediates FFA uptake. It is not clear whether FFA- and proteinuria-mediated CD36 activation coordinates NLRP3 inflammasomes to induce renal tubular injury and inflammation. In this study, we investigated the roles of CD36 and NLRP3 inflammasomes in FFA-induced renal injury in high-fat diet (HFD)-induced obesity. HFD-fed C57BL/6 mice and palmitate-treated HK2 renal tubular cells were used as in vivo and in vitro models. Immunohistochemical staining showed that CD36, IL-1β, and IL-18 levels increased progressively in the kidneys of HFD-fed mice. Sulfo- N-succinimidyl oleate (SSO), a CD36 inhibitor, attenuated the HFD-induced upregulation of NLRP3, IL-1β, and IL-18 and suppressed the colocalization of NLRP3 and ASC in renal tubular cells. In vitro, SSO abolished the palmitate-induced activation of IL-1β, IL-18, and caspase-1 in HK2 proximal tubular cells. Furthermore, treatment with SSO and the knockdown of caspase-1 expression by siRNA both inhibited palmitate-induced cell death and apoptosis in HK2 cells. Collectively, palmitate causes renal tubular inflammation, cell death, and apoptosis via the CD36/NLRP3/caspase-1 axis, which may explain, at least in part, the mechanism underlying FFA-related renal tubular injury. The blockade of CD36-induced cellular processes is therefore a promising strategy for treating obesity-related nephropathy.

Keyword: metabolic syndrome

Acute hypoxic preconditioning prevents -induced cardiomyocyte apoptosis via switching GLUT4-glucose pathway back to CD36-fatty dependent.

is a risk factor for the development of cardiovascular diseases. Myocardial cell damage leads to an imbalance of energy metabolism, and many studies have indicated that short-term hypoxia during myocardial cell injury has a protective effect. In our previous animal studies, we found that short-term hypoxia in the heart has a protective effect, but long-term hypoxia increases myocardial cell injury. (PA)-treated H9c2 cardiomyoblasts and neonatal rat ventricle cardiomyocytes were used to simulate hyperlipidemia model, which suppress cluster of differentiation 36 (CD36) and activate glucose transporter type 4 (GLUT4). We exposed the cells to short- and long-term hypoxia and investigated the protective effects of hypoxic preconditioning on PA-induced lipotoxicity in H9c2 cardiomyoblasts and neonatal rat cardiomyocytes. Preconditioning with short-term hypoxia enhanced both CD36 and GLUT4 metabolism pathway protein levels. Expression levels of phospho-PI3K, phospho-Akt, phospho-AMPK, SIRT1, PGC1α, PPARα, CD36, and CPT1β induced by PA was reversed by short-term hypoxia in a time-dependent manner. PA-induced increased GLUT4 membrane protein level was reduced in the cells exposed to short-term hypoxia and si-PKCζ. Short-term hypoxia, resveratrol and si-PKCζ rescue H9c2 cells from apoptosis induced by PA and switch the pathway from GLUT4 dependent to CD36 dependent. We demonstrate short-term hypoxic preconditioning as a more efficient way as resveratrol in maintaining the energy metabolism of hearts during hyperlipidemia and can be used as a therapeutic strategy.© 2017 Wiley Periodicals, Inc.

Keyword: metabolic syndrome

Novel proapoptotic effect of hepatocyte growth factor: synergy with palmitate to cause pancreatic {beta}-cell apoptosis.

Increasing evidence suggests that elevation of plasma fatty acids that often accompanies insulin resistance contributes to beta-cell insufficiency in obesity-related type 2 diabetes. Circulating levels of hepatocyte growth factor (HGF) are increased in humans with and obesity. HGF is known to protect beta-cells against streptozotocin and during islet engraftment. However, whether HGF is a beta-cell prosurvival factor in situations of excessive lipid supply has not been deciphered. Mice overexpressing HGF in the beta-cell [rat insulin type II promoter (RIP)-HGF transgenic mice] fed with standard chow display improved glucose homeostasis and increased beta-cell mass and proliferation compared with normal littermates. However, after 15 wk of high-fat feeding, glucose homeostasis and beta-cell expansion and proliferation are indistinguishable between normal and transgenic mice. Interestingly, RIP-HGF transgenic mouse beta-cells and normal beta-cells treated with HGF display increased sensitivity to palmitate-mediated apoptosis in vitro. Palmitate completely eliminates Akt and Bad phosphorylation in RIP-HGF transgenic mouse islets. HGF-overexpressing islets also show significantly decreased AMP-activated protein kinase-alpha and acetyl-coenzyme A carboxylase phosphorylation, diminished fatty oxidation, increased serine palmitoyltransferase expression, and enhanced ceramide formation compared with normal islets. Importantly, human islets overexpressing HGF also display increased beta-cell apoptosis in the presence of palmitate. Treatment of both mouse and human islet cells with the de novo ceramide synthesis inhibitors myriocin and fumonisin B1 abrogates beta-cell apoptosis induced by HGF and palmitate. Collectively, these studies indicate that HGF can be detrimental for beta-cell survival in an environment with excessive fatty supply.

Keyword: metabolic syndrome

Use of stable isotope labeling technique and mass isotopomer distribution analysis of [(13)C]palmitate isolated from surfactant disaturated phospholipids to study surfactant in vivo kinetics in a premature infant.

Pulmonary surfactant is a complex mixture of phospholipids and proteins which lowers surface tension and maintains alveolar expansion at end expiration. Developmental and genetic disruption of pulmonary surfactant metabolism leads to respiratory distress in newborns. Stable isotope labeling of precursors of disaturated phospholipids, the most abundant and specific component of pulmonary surfactant, permits the measurement of the kinetics of surfactant metabolism in vivo. We measured [U-(13)C(6)]glucose incorporation into derived from disaturated surfactant phospholipids. A 24 h infusion of [U-(13)C(6)]glucose (140 mg kg(-1)) was administered to a premature infant who required mechanical ventilation for respiratory distress ; tracheal aspirate samples were obtained at the start of the infusion and at regular intervals for the next 70 h. Each tracheal aspirate sample was incubated with osmium tetroxide to isolate disaturated surfactant phospholipids. Methyl esters of the fatty acids in the disaturated phospholipids were prepared and the enrichment of [(13)C]methyl palmitate was measured by gas chromatography/mass spectrometry (GC/MS) and gas chromatography/combination/isotope ratio mass spectrometry (GC/C/IRMS). Mass isotopomer distribution analysis (MIDA) was used to calculate the fractional synthetic rate (FSR) of palmitate synthesized from acetate. With both GC/MS and GC/C/IRMS, palmitate (13)C enrichment was first detected 12.3 h after the start of the tracer infusion. The enrichment increased in a linear fashion, reached a peak at 47 h and remained constant in the remainder of the samples. The FSR of palmitate from acetate was 5.2% per day. Stable isotope techniques and MIDA will provide insights into the kinetics of surfactant metabolism in newborns with respiratory dysfunction.Copyright 2000 John Wiley & Sons, Ltd.

Keyword: metabolic syndrome

[The person in philogenesis is not (omnivores), but the herbivores with the carnivores past and the fuzzy future. Biological function of trophology (nutrition) in ontogenesis.]

According to the phylogenetic theory of general pathology, seven biological functions have been formed over billions of years. 1. biological function of trophology, nutrition; 2. homeostasis function; 3. biological function of endoecology; 4. function of adaptation; 5. function of the continuation of the species; 6. function of locomotion and 7. cognitive biological function, including intelligence. Millions of years in life consistently in the waters of several oceans, all the ancestors of man were carnivorous (Carnivores), fish-eating mammals. When the ocean retreated and the carnivorous (fish-eating) were on land, each individual privatized a "piece" of the ocean. Animals transformed it ito a pool of intercellular medium in vivo. The biological role of the late in the phylogeny of insulin is the formation of new biological functions in vivo. The action of insulin has transformed the carnivorous (fish-eating) ocean into herbivorous (Herbivores) species on land. There was it by synthesis in vivo from exogenous glucose of fatty acids (FA). Regulatory action of insulin was the directed conversion of exogenous glucose into ω-6 C18: 1 cis-oleic FA. Insulin late in phylogeny expressed the synthesis of new, conjugated enzymes: it is palmitoyl-CoA-elongase and stearyl-CoAdesaturase. Two enzymes synthesized FAs along the way: synthesized in situ de novo, from exogenous glucose, C16: 0 → C18: 0 stearic → ω-6 C18: 1 cis-oleic without accumulation of stearic FA. Insulin is not converted into an oleic FA exogenous from carnivorous food. On land, the action of insulin transformed the species Homo sapiens, into a herbivore, but with carnivorous, fish-eating, past. The idea of a person as omnivorous (Omnivor) - nonsense; such forms of nature did not form. Violation of the function of nutrition, the biological reaction of exotrophy (external nutrition), is the etiological and pathogenetic basis of the seven pandemics, the diseases of civilization. 1. Atherosclerosis and atheromatosis; 2. arterial hypertension; 3. ; 4. obesity; 5. of insulin resistance; 6. non-alcoholic fatty liver disease and 7. endogenous hyperuricemia. The primary prevention of pandemics in the biological function of nutrition, in the biological reactions of exo-and endotrophy, will allow us to understand the theoretical bases and implementation of preventive actions that will determine the characteristics of nutrition in the future.

Keyword: metabolic syndrome

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to . NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic . Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: metabolic syndrome

Chitosan Oligosaccharides Improve Glucolipid Metabolism Disorder in Liver by Suppression of Obesity-Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid metabolism disorder using (PA)-induced HepG2 cells and high-fat diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated lipid accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid metabolism-related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid metabolism disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid metabolism-related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid metabolism disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid metabolism-related diseases.

Keyword: metabolic syndrome

Characterization of L-aminocarnitine, an inhibitor of fatty oxidation.

The pathogenesis of hypoketotic hypoglycemia and cardiomyopathy in patients with fatty oxidation (FAO) disorders is still poorly understood. In vitro studies are hampered by the lack of natural mutants to asses the effect of FAO inhibition. In addition, only a few inhibitors of FAO are known. Furthermore, most inhibitors of FAO are activating ligands of peroxisome proliferator-activated receptors (PPARs). We show that l-aminocarnitine (L-AC), a carnitine analog, inhibits FAO efficiently, but does not activate PPAR. L-AC inhibits carnitine palmitoyltransferase (CPT) with different sensitivities towards CPT1 and CPT2, as well as carnitine acylcarnitine translocase (CACT). We further characterized L-AC using fibroblasts cell lines from controls and patients with different FAO defects. In these cell lines acylcarnitine profiles were determined in culture medium after loading with [U-(13)C]. In control fibroblasts, L-AC inhibits FAO leading to a reduction of C2-acylcarnitine and elevation of C16-acylcarnitine. In very long-chain acyl-CoA dehydrogenase (VLCAD)-deficient fibroblasts, L-AC decreased the elevated C14-acylcarnitine and increased C16-acylcarnitine. In CACT and CPT2-deficient cell lines, L-AC did not change the already elevated C16-acylcarnitine level, showing that CPT1 is not inhibited. Oxidation of pristanic was only partly inhibited at high L-AC concentrations, indicating minimal CACT inhibition. Therefore, we conclude that in intact cells L-AC inhibits CPT2. Combined with our observation that l-AC does not activate PPAR, we suggest that L-AC is useful to simulate a FAO defect in cells from different origin.

Keyword: metabolic syndrome

Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with .

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an energy-restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with .The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of .After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk energy-restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

increases pro-oxidant adaptor protein p66Shc expression and affects vascularization factors in angiogenic mononuclear cells: Action of resveratrol.

A defect in neo-vascularization process involving circulating angiogenic mononuclear cells (CACs) dysfunction is associated with diabetes. We showed that oxidative stress was elevated in CACs cultured from blood of individuals with (MetS) and diabetes. We then assessed the action of (PA), a deregulated and increased NEFA in disorders, focusing on its oxidant potential. We observed that the phyto-polyphenol resveratrol normalized oxidative stress both in CACs isolated from MetS patients or treated with PA. Resveratrol further decreased the deleterious action of PA on gene expression of vascularization factors (TNFα, VEGF-A, SDF1α, PECAM-1, VEGFR2, Tie2 and CXCR4) and improved CAC motility. Particularly, resveratrol abolished the PA-induced over-expression of the pro-oxidant protein p66Shc. Neither KLF2 nor SIRT1, previously shown in resveratrol and p66Shc action, was directly involved. Silencing p66Shc normalized PA action on VEGF-A and TNFα specifically, without abolishing the PA-induced oxidative stress, which suggests a deleterious role of p66Shc independently of any major modulation of the cellular oxidative status in a high NEFA levels context. Besides showing that resveratrol reverses PA-induced harmful effects on human CAC function, certainly through profound cellular modifications, we establish p66Shc as a major therapeutic target in disorders, independent from glycemic control.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Plasma fatty composition, estimated desaturase activities, and their relation with the in a population at high risk of cardiovascular disease.

The (MetS) is a clustering of various abnormalities which is associated with increased risk of cardiovascular disease (CVD) and type 2 diabetes mellitus. Due to its increasing prevalence, it has become an important public health concern. Altered fatty (FA) composition and desaturase activities have been associated with several diseases, including MetS. The aim of the present study was to evaluate the relationship of the plasma FA profile and desaturase activities with the MetS in a Mediterranean population at high risk of CVD.Baseline data from 427 participants aged 55-80 years who took part in the interventional PREDIMED study were obtained. Individual FA was determined in plasma and desaturase activities were estimated from product/precursor ratios. Odds ratios (OR) and partial correlation coefficients were used to examine these relations with MetS and its components, respectively.We found higher levels of C14:0, C16:0, C16:1n-7, estimated Δ(9)- or stearoyl-CoA desaturase (SCD), and estimated Δ(6) desaturase (D6D), and lower levels of C18:2n-6 in people with MetS compared to those without it. After adjustment for several confounders, only higher quartiles of C14:0, C16:0, C16:1n-7, and D6D were found to be associated with an increasing prevalence of MetS, while higher quartiles of C18:2n-6 were inversely associated with MetS. High proportions of C14:0, C16:0, C16:1n-7, C20:3n-6, SCD, and D6D, and decreased proportions of C18:2n-6 and estimated Δ(5)-desaturase (D5D) were associated with adverse profiles of several risk factors. Women showed more unhealthy FA pattern and lipid profiles than men, but only among those with MetS.A FA composition and estimated desaturase activities consisting in high levels of SFA, SCD and D6D, and low levels of PUFA and D5D are associated with increased MetS probability and are characteristic of people presenting MetS, especially women. These findings support those observed in non-Mediterranean populations in which an altered FA profile and estimated desaturase activities are associated with MetS.Copyright © 2013 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Keyword: metabolic syndrome

Polysaccharide from Rubus chingii Hu affords protection against -induced lipotoxicity in human hepatocytes.

(PA) is known to induce lipotoxicity, a as a result of lipid accumulation in multiple cell lines. Bioactive phytochemicals derived from vegetables and fruits have gained increasing attention owing to their potential on suppressing the detrimental effect of excessive PA accumulation. However, the protective effect of natural phytochemicals derived from Rubus chingii Hu, a kind of fruit widely grown in China, against PA-induced lipotoxicity is still uncleared. In the present study, we therefore extracted the polysaccharide from Rubus chingii Hu, and identified its chemical structure. Structural characterization by HPLC, HPGPC, IR spectroscopy and GC indicated that the polysaccharide mainly consists of galacturonic and arabinose with copious 1\u202f→\u202f2 glycosidic linkages in its backbone. In addition, our results showed the cytoprotective effect of the polysaccharide against PA-induced lipotoxicity in normal human hepatocyte cell line L02. Further study indicated that the polysaccharide mitigated oxidative stress through impeding cellular reactive oxygen species (ROS) accumulation, alleviating mitochondrial membrane potential (MMP) collapse and attenuating glutathione (GSH) reduction. Overall, this study revealed that Rubus chingii Hu polysaccharide was capable of effectively alleviating -induced lipotoxicity, which provided a novel perspective of the health-promoting potential of isolated polysaccharide.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Effects of 1α,25 Dihydroxyvitamin D on Pro-inflammatory Cytokines of Treated Thp-1 Cells.

The level of saturated acids, such as (PA), correlates with chronic inflammation in obese and metabolic syndrome patients. However, low level of vitamin D is observed in those conditions. The aim of this study is to investigate effects of 1α,25(OH) D on PA-treated THP-1 cells. Using quantitative real-time polymerase chain reaction, we measure mRNA expression of pro-inflammatory cytokines: TNF-α, Interleukin (IL)-1β, IL-6, and chemokine IL-8 under PA and 1α,25(OH) D influence. PA, at all concentrations (25-100 μM), enhanced LPS stimulatory effect on those mRNA expression compared to LPS-treated and -untreated cells. Combination with 1α,25(OH) D increased cytokine expression at high (10 M) and high-normal (10 M) concentrations compared to PA + LPS and LPS alone, both for 2 and 24 h. However, low-normal (10 M) and low (10 M) levels of 1α,25(OH) D could not enhance PA effect, but mRNA expression of pro-inflammatory cytokine was higher than LPS-treated cells. Upstream pathway of 1α,25(OH) D , which is cholecalciferol, also gave the similar result. Further, inhibition of calcium pathway does not play a role in this mechanism. Thus, these findings support pro-inflammatory effect of PA and vitamin D on innate immune response, especially on fat-induced inflammation.The effect of vitamin D on chronic inflammation in obesity is uncertain. This study shows an in vitro possibility that vitamin D could exaggerate inflammation when combined with high SFAs. The idea of using vitamin D supplement to modulate inflammation in fat-related inflammation needs further refined experiments before its clinical application.© 2017 Institute of Food Technologists®.

Keyword: metabolic syndrome

[The positional isomers of triglycerides in oils, fats and apoB-100 lipoproteins: and oleic modes of metabolism of fatty acids-substrates for energy acquiring].

Even total resemblance of content of fatty acids in triglycerides has both no standing for their functional unity nor even identity of their physical chemical characteristics. The etherification of fatty acids in various positions of three-atomic glycerin separates triglycerides on and oleic substrates for energy acquiring by cells. The kinetic parameters of biochemical reactions under mode of metabolism of fatty acids are always low. The myocytes in biological reaction of exotrophy experience deficiency of exogenous fatty acids which in vivo is to permanently supply through activation of biological reaction of endotrophy--enhancement of lipolysis in adipocytes. The biological role of insulin is to prevent formation in vivo of mode of metabolism of saturated and monoenic fatty acids. Under this condition, the necessity to activate lipolysis and to increase in blood plasma concentration of unesteritied fatty acids forms of resistance to insulin. The surplus of fatty in food and deficiency of insulin show in vivo unidirectional a physiologic action. The formation of mode of metabolism of energy substrates--portion of pathogenesis of atherosclerosis, , obesity, non-alcoholic fatty infiltration of liver and partiallly essential arterial hypertension.

Keyword: metabolic syndrome

Plasma biomarkers for of phlegm and blood stasis in hyperlipidemia and atherosclerosis.

To explore the plasma metabolite profiles in patients with the of phlegm and blood stasis in hyperlipidemia and atherosclerosis (As), and to search for the biomarkers of the .The plasma metabolite profiles of 31 patients with the of phlegm and blood stasis in hyperlipidemia and As, 6 patients with syndromes without phlegm and blood stasis, and 10 healthy subjects were analyzed by gas chromatography-mass spectrometry (GC-MS). Partial least squares-discriminant analyses (PLS-DA) were used to carry out the pattern-recognition analyses of the data. The plasma biomarkers of patients were obtained by variable importance plot value (VIP value) and Student\'s t-test. The structures of biomarkers were defined by the National Institute of Standards and Technology (NIST) database.PLS-DA score plots of plasma metabolomes did not show overlap between the phlegm-blood stasis group and syndromes without phlegm and blood stasis group, whereas significant differences in the concentrations in the plasma of 5 metabolites were found (P < 0.05). They were identified as urine, isoleucine, glucuronic , and glycerol by searching in NIST database. The concentrations of four metabolites in the plasma of patients with of phlegm and blood stasis were higher than those with syndromes without phlegm and blood stasis, whereas the glycerol concentration was lower.Compared with patients with syndromes without phlegm and blood stasis, five metabolites showed abnormal levels in patients with the of phlegm and blood stasis. These metabolites could be diagnostic and prognostic biomarkers.

Keyword: metabolic syndrome

Increased lipid synthesis and decreased β-oxidation in the liver of SHR/NDmcr-cp (cp/cp) rats, an animal model of .

SHR/NDmcr-cp (cp/cp) rats (SHR/NDcp) are an animal model of . A previous study of ours revealed drastic increases in the mass of (16:0), oleic (18:1n-9), palmitoleic (16:1n-7), cis-vaccenic (18:1n-7) and 5,8,11-eicosatrienoic acids in the liver of SHR/NDcp. However, detailed information on the class of lipid accumulated and the mechanism responsible for the overproduction of the accumulated lipid in the liver was not obtained. This study aimed to characterize the class of lipid accumulated and to explore the mechanism underlying the lipid accumulation in the liver of SHR/NDcp, in comparison with SHR/NDmcr-cp (+/+) (lean hypertensive littermates of SHR/NDcp) and Wistar Kyoto rats. In the liver of SHR/NDcp, de novo synthesis of fatty acids (16:0, 18:1n-9 and 16:1n-7) and triacylglycerol (TAG) synthesis were up-regulated and fatty β-oxidation was down-regulated. These perturbations of lipid metabolism caused fat accumulation in hepatocytes and accumulation of TAG, which were enriched with 16:0, 18:1n-9 and 16:1n-7, in the liver of SHR/NDcp. On the other hand, no changes were found in hepatic contents of diacylglycerol and unesterified fatty (FFA); among FFA, there were no differences in the hepatic concentrations of unesterified 16:0 and stearic between SHR/NDcp and two other groups of rats. Moreover, little change was brought about in the expression of genes responsive to endoplasmic reticulum stress in the liver of SHR/NDcp. These results may reinforce the pathophysiological role of stearoyl-CoA desaturase 1 and fatty elongase 6 in the liver of SHR/NDcp.

Keyword: metabolic syndrome

[The physical chemical and biological features of triglycerides. The cell absorption of functionally different +oleic lipoproteins of very low and density and linoleic+linolenic lipoproteins of low density.]

The earlier insulin-independent low-density lipoproteins and more late insulin-dependent very low-density lipoproteins implement different functions at the stages of phylogenesis. The disorder of biological function of trophology, alteration of fatty acids in triglycerides, prevalence of very low-density lipoproteins over oleic very low-density lipoproteins supply mitochondria of cells with non-optimal substrate - saturated fatty for gaining energy, ATP synthesis. Physiologically, cells implement oleic alternative of fatty acids metabolism, oxidizing mainly ω-9 endogenous oleic mono-unsaturated fatty . The pathology of low density lipoproteins is primary deficiency of poly-unsaturated fatty acids in cells, atherosclerosis and atheromotosis of intima of arteries of elastic type with development of dense plaques from poly-unsaturated fatty acids in the form of polyethers of cholesterol. The pathology of very low-density lipoproteins includes: a) of resistance to insulin; b) pathology of phylogenetically earlier insulin-independent visceral fatty tissue - ; c) pathology of phylogenetically later insulin-dependent subcutaneous adipocytes - obesity; d) secondary atherosclerosis, under cumulation of low-density lipoproteins in blood with development of atherothrombosis of intima of arteries, soft plaques rich with triglycerides. As for the prevention of disorders of transfer of fatty acids to very low-density lipoproteins and low-density lipoproteins is common in many ways - minimization of aphysiological effect of surplus amount of food, biological function of diet. The prevention at the level of population includes: a) maximal limitation of content of saturated fatty in food; b) moderate increasing of polysaturated fatty acids, ω-3 poly-saturated fatty acids predominantly; c) increasing of physical activity. The pharmaceuticals are not provided by biology in primary prevention of pandemics under aphysiological impact of environment factors.

Keyword: metabolic syndrome

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: metabolic syndrome

Saturated fatty acids induce development of both and osteoarthritis in rats.

The predominant saturated fatty acids (SFA) in human diets are lauric (LA, C12:0), myristic (MA, C14:0), (PA, C16:0) and stearic (SA, C18:0). The aim of this study was to investigate whether diets containing individual SFA together with excess simple carbohydrates induce osteoarthritis (OA)-like changes in knee joints and signs of in rats. Rats were given either a corn starch diet or a diet composed of simple carbohydrates together with 20% LA, MA, PA, SA or beef tallow for 16 weeks. Rats fed beef tallow, SA, MA or PA diets developed signs of , and also exhibited cartilage degradation and subchondral bone changes similar to OA. In contrast, replacement of beef tallow with LA decreased signs of together with decreased cartilage degradation. Furthermore, PA and SA but not LA increased release of matrix sulphated proteoglycans in cultures of bovine cartilage explants or human chondrocytes. In conclusion, we have shown that longer-chain dietary SFA in rats induce both and OA-like knee changes. Thus, diets containing SFA are strongly relevant to the development or prevention of both OA and .

Keyword: metabolic syndrome

[The cognitive biological function, difference in etiology and generality of pathogenesis of pandemics. The metabolism regulation in carnivorous and herbivorous animals under effect of environment factors.]

In phylogenesis, carnivorous animals initially and later herbivorous animals formed separate transfer of exogenous fatty acids in blood, predominantly and later - endogenous oleic pool, synthesized in hepatocytes from glucose. In vivo, exogenous fatty acids transfer α-lipoproteins to all cells and chylomicrons to hepatocytes. The endogenous oleic fatty , synthesized in hepatocytes is transferred to all cells by β-lipoproteins and to insulin-dependent lipoproteins of very low density. The limitation of cognitive biological function in herbivorous animals underlies the process of transferring of surplus amount of exogenous and alcohol cholesterol by β-lipoproteins becoming a non-physiological one. The bio-availability is affected concerning cells of exogenous polyene fatty acids that transfer in blood α-lipoproteins in carnivorous animals and at the first stage in herbivorous animals. The formation of non-ligand very low density lipoproteins-β-proteins is affected and their collection and partial utilization in intima generates lipoidosis. In summary, all this becomes a cause of formation of atherosclerosis and atheromatosis. The limitations in phylogenesis of cognitive biological function are considered as an etiological factor of pandemics. The etiological factors are specific for every one of pandemics and pathogenesis of all pandemics is developed following common algorithm. It is likely that pandemics (atherosclerosis and atheromatosis, essential arterial hypertension, , of insulin resistance, obesity and non-alcoholic fatty disease of liver)are quite not identical to nosologic forms of diseases if considered according etiological factors, pathogenesis and pathology rate in population. The exclusion of all inherent forms of disorder of metabolism of fatty acids and their transfer in β-lipoproteins demonstrates that problem of atherosclerosis, atheromatosis, coronarosclerosis and ischemic heart disease mainly turns out to be a case of eaten one\'s fill , a result of environment factors impact, disorder of biological function of trophology, biological reaction of exotrophy (external nutrition) and biological function of homeostasis. The prevention of atherosclerosis, atheromatosis and ischemic heart disease in population will be effective only under condition of obligatory individual support of actioning cognitive biological function by more complete biological function of intelligence. The active prevention of pandemics by force of personal development of cognitive biological function, intelligence function, provides no application of pharmaceuticals. The disorder of cognitive biological function is needed to be improved, incorrect positioning of Homo Sapiens species as omnivorous in animal world; Homo Sapiens is a herbivorous one.

Keyword: metabolic syndrome

exerts pro-inflammatory effects on vascular smooth muscle cells by inducing the expression of C-reactive protein, inducible nitric oxide synthase and tumor necrosis factor-α.

Atherosclerosis is a chronic inflammatory disease in the vessel, and inflammatory cytokines play an important role in the inflammatory process of atherosclerosis. A high level of free fatty acids (FFAs) produced in lipid metabolism disorders are known to participate in the formation of atherosclerosis through multiple bioactivities. As the main saturated fatty in FFAs, stimulates the expression of inflammatory cytokines in macrophages. However, it is unclear whether exerts a pro-inflammatory effect on vascular smooth muscle cells (VSMCs). The purpose of the present study was to observe the effect of on the expression of C-reactive protein (CRP), tumor necrosis factor α (TNF-α) and inducible nitric oxide synthase (iNOS) in VSMCs. Rat VSMCs were cultured, and was used as a stimulant for CRP, TNF-α and iNOS expression. mRNA expression was assayed with reverse transcription-polymerase chain reaction, and protein expression was detected with western blot analysis and immunocytochemistry. The results showed that significantly stimulated mRNA and protein expression of CRP, TNF-α and iNOS in VSMCs in time- and concentration-dependent manners, and therefore, is able to exert a pro-inflammatory effect on VSMCs via stimulating CRP, TNF-α and iNOS expression. The findings provide a novel explanation for the direct pro-inflammatory and atherogenic effects of , and for the association with , such as type 2 diabetes mellitus, obesity and atherosclerosis. Therefore, the intervention with anti-inflammatory agents may effectively delay the formation and progression of atherosclerosis in patients with .

Keyword: metabolic syndrome

Serum metabolomics study of polycystic ovary based on UPLC-QTOF-MS coupled with a pattern recognition approach.

Metabolomics has become an important tool in distinguishing changes in pathways and the diagnosis of human disease. Polycystic ovary (PCOS) is a relatively complicated, heterogeneous endocrine disorder. The etiology and pathogenesis of PCOS remain uncertain. In this study, based on the platform of ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and the method of pattern recognition, a comprehensive metabolomics approach has been applied to explore the changes in profiling between PCOS patients (n\u2009=\u200920) and controls (n\u2009=\u200915) as well as insulin-resistance (IR) PCOS patients (n\u2009=\u200911) and non-IR PCOS subjects (n\u2009=\u20099) in serum. In total, 36 metabolites were found significantly different between PCOS and controls, and 9 metabolites were discovered significantly different between IR and non-IR PCOS patients. Significant increases in the levels of saturated and unsaturated fatty acids (myristic , linoleic , 9-/13-HODE, etc.), fatty amides ( amide, oleamide), dehydroepiandrosterone sulfate, L-glutamic , azelaic , L-glyceric , pyroglutamic , and decreases in the levels of lysophosphatidylethanolamine, lysophosphatidylcholine, uridine, and L-carnitine were found in PCOS patients compared with controls. In IR PCOS patients, linoleic , myristic , palmitoleic , and vaccenic also increased significantly compared with non-IR PCOS patients. All these changed metabolites showed abnormalities of steroid hormone biosynthesis, amino acids and nucleosides metabolism, glutathione metabolism, and lipids and carbohydrates metabolism in PCOS patients. The subgroup IR PCOS patients exhibited greater deviations than non-IR PCOS patients. These findings may help yield promising insights into the pathogenesis and advance the diagnosis and prevention of PCOS. Graphical Abstract Serum metabolomics signature of polycystic ovary .

Keyword: metabolic syndrome

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the liver of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in liver cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the liver steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and liver lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: metabolic syndrome

Polyunsaturated fatty acids effect on serum triglycerides concentration in the presence of components. The Alaska-Siberia Project.

Serum fatty acids (FAs) have wide effects on metabolism: Serum saturated fatty acids (SFAs) increase triglyceride (TG) levels in plasma, whereas polyunsaturated fatty acids (PUFAs) reduce them. Traditionally, Eskimos have a high consumption of omega-3 fatty acids (omega3 FAs); but the Westernization of their food habits has increased their dietary SFAs, partly reflected in their serum concentrations. We studied the joint effect of serum SFAs and PUFAs on circulating levels of TGs in the presence of components. We included 212 men and 240 women (age, 47.9 +/- 15.7 years; body mass index [BMI], 26.9 +/- 5.3) from 4 villages located in Alaska for a cross-sectional study. Generalized linear models were used to build surface responses of TG as functions of SFAs and PUFAs measured in blood samples adjusting by sex, BMI, and village. The effects of individual FAs were assessed by multiple linear regression analysis, and partial correlations (r) were calculated. The most important predictors for TG levels were glucose tolerance (r = 0.116, P = .018) and BMI (r = 0.42, P < .001). Triglyceride concentration showed negative associations with 20:3omega6 (r = -0.16, P = .001), 20:4omega6 (r = -0.14, P = .005), 20:5omega3 (r = -0.17, P < .001), and 22:5omega3 (r = -0.26, P < .001), and positive associations with (r = 0.16, P < .001) and 18:3omega3 (r = 0.15, P < .001). The surface response analysis suggested that the effect of on TG is blunted in different degrees according to the PUFA chemical structure. The long-chain omega3, even in the presence of high levels of saturated fat, was associated with lower TG levels. Eicosapentaenoic (20:5omega3) had the strongest effect against on TG. The total FA showed moderate association with levels of TG, whereas SFA was positively associated and large-chain PUFA was negatively associated. The Westernized dietary habits among Eskimos are likely to change their profile and increase comorbidities related to disease.

Keyword: metabolic syndrome

Anxa2 gene silencing attenuates obesity-induced insulin resistance by suppressing the NF-κB signaling pathway.

Insulin resistance (IR) continues to pose a major threat to public health due to its role in the pathogenesis of and its ever-increasing prevalence on a global scale. The aim of the current study was to investigate the efficacy of Anxa2 in obesity-induced IR through the mediation of the NF-κB signaling pathway. Microarray analysis was performed to screen differentially expressed genes associated with obesity. To verify whether Anxa2 was differentially expressed in IR triggered by obesity, IR mouse models were established in connection with a high-fat diet (HFD). In the mouse IR model, the role of differentially expressed Anxa2 in glycometabolism and IR was subsequently detected. To investigate the effect of Anxa2 on IR and its correlation with inflammation, a (PA)-induced IR cell model was established, with the relationship between Anxa2 and the NF-κB signaling pathway investigated accordingly. Anxa2 was determined to be highly expressed in IR. Silencing Anxa2 was shown to inhibit IR triggered by obesity. When Anxa2 was knocked down, elevated expression of phosphorylated insulin receptor substrate 1 (IRS1), IRS1 and peroxisome proliferator-activated receptor coactivator-1a, and glucose tolerance and insulin sensitivity along with 2-deoxy-d-glucose uptake was detected, whereas decreased expression of suppressor of cytokine signaling 3, IL-6, IL-1β, TNF-α, and p50 was observed. Taken together, the current study ultimately demonstrated that Anxa2 may be a novel drug strategy for IR disruption, indicating that Anxa2 gene silencing is capable of alleviating PA or HFD-induced IR and inflammation through its negative regulatory role in the process of p50 nuclear translocation of the NF-κB signaling pathway.

Keyword: metabolic syndrome

Reduced levels of SCD1 accentuate palmitate-induced stress in insulin-producing β-cells.

Stearoyl-CoA desaturase 1 (SCD1) is an ER resident enzyme introducing a double-bond in saturated fatty acids. Global knockout of SCD1 in mouse increases fatty oxidation and insulin sensitivity which makes the animal resistant to diet-induced obesity. Inhibition of SCD1 has therefore been proposed as a potential therapy of the . Much of the work has focused on insulin target tissue and very little is known about how reduced levels of SCD1 would affect the insulin-producing β-cell, however. The aim of the present study was therefore to investigate how reduced levels of SCD1 affect the β-cell.Insulin-secreting MIN6 cells with reduced levels of SCD1 were established by siRNA mediated knockdown. When fatty oxidation was measured, no difference between cells with reduced levels of SCD1 and mock-transfected cells were found. Also, reducing levels of SCD1 did not affect insulin secretion in response to glucose. To investigate how SCD1 knockdown affected cellular mechanisms, differentially regulated proteins were identified by a proteomic approach. Cells with reduced levels of SCD1 had higher levels of ER chaperones and components of the proteasome. The higher amounts did not protect the β-cell from palmitate-induced ER stress and apoptosis. Instead, rise in levels of p-eIF2α and CHOP after palmitate exposure was 2-fold higher in cells with reduced levels of SCD1 compared to mock-transfected cells. Accordingly, apoptosis rose to higher levels after exposure to palmitate in cells with reduced levels of SCD1 compared to mock-transfected cells.In conclusion, reduced levels of SCD1 augment palmitate-induced ER stress and apoptosis in the β-cell, which is an important caveat when considering targeting this enzyme as a treatment of the .

Keyword: metabolic syndrome

induces production of proinflammatory cytokine interleukin-8 from hepatocytes.

Obesity and the are closely correlated with hepatic steatosis. Simple hepatic steatosis in nonalcoholic fatty liver disease can progress to nonalcoholic steatohepatitis (NASH), which can be a precursor to more serious liver diseases, such as cirrhosis and hepatocellular carcinoma. The pathogenic mechanisms underlying progression of steatosis to NASH remain unclear; however, inflammation, proinflammatory cytokines, and oxidative stress have been postulated to play key roles. We previously reported that patients with NASH have elevated serum levels of proinflammatory cytokines, such as interleukin-8 (IL-8), which are likely to contribute to hepatic injury. This study specifically examines the effect of hepatic steatosis on IL-8 production. We induced lipid accumulation in hepatocytes (HepG2, rat primary hepatocytes, and human primary hepatocytes) by exposing them to pathophysiologically relevant concentrations of to simulate the excessive influx of fatty acids into hepatocytes. Significant fat accumulation was documented morphologically by Oil Red O staining in cells exposed to , and it was accompanied by an increase in intracellular triglyceride levels. Importantly, was found to induce significantly elevated levels of biologically active neutrophil chemoattractant, IL-8, from steatotic hepatocytes. Incubation of the cells with palmitate led to increased IL-8 gene expression and secretion (both mRNA and protein) through mechanisms involving activation of nuclear factor kappaB (NF-kappaB) and c-Jun N-terminal kinase/activator protein-1.These data demonstrate for the first time that lipid accumulation in hepatocytes can stimulate IL-8 production, thereby potentially contributing to hepatic inflammation and consequent liver injury.

Keyword: metabolic syndrome

Increased erythrocytes n-3 and n-6 polyunsaturated fatty acids is significantly associated with a lower prevalence of steatosis in patients with type 2 diabetes.

Non-alcoholic fatty liver disease (NAFLD) is commonly associated with obesity, and type 2 diabetes. Although dietary fat contributes substantially to the accumulation of liver fat, the role of individual fatty acids in this accumulation is unclear.In this study, we set out to determine whether liver fat content (LFC), was associated with red blood cell fatty (RBC-FA) composition in people with type 2 diabetes.One hundred and sixty-two type 2 diabetic patients were included in this study. LFC was measured using (1)H-MR Spectroscopy. RBC-FA composition was measured by gas chromatography.One hundred and nine (67.2%) patients had steatosis. Patients with steatosis had a higher BMI (p = 0.0005), and higher plasma triglyceride levels (p = 0.009) than did patients without steatosis. We report a significant association between (16:0), palmitoleic (16:1n-7) concentrations and ratio of monounsaturated to saturated fatty (palmitoleic to ) and higher liver fat content. Total polyunsaturated fatty (PUFA), homo-gamma-linolenic (20:3n-6), docosahexaenoic (22:6n-3), and arachidonic (20:4 n-6) were associated with lower LFC.Our data showed that an increased erythrocytes long-chain n-3 and n-6 fatty acids was associated with a lower prevalence of steatosis in patients with type 2 diabetes. These results suggest that n-3 and n-6 fatty acids supplementation could be a promising treatment for NAFLD in patients with type 2 diabetes.Copyright © 2011 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Keyword: metabolic syndrome

Resveratrol reduces intracellular reactive oxygen species levels by inducing autophagy through the AMPK-mTOR pathway.

Oxidative stress induced by free fatty aggravates endothelial injury, which leads to diabetic cardiovascular complications. Reduction of intracellular oxidative stress may attenuate these pathogenic processes. The dietary polyphenol resveratrol reportedly exerts potential protective effects against endothelial injury. This study determined whether resveratrol can reduce the (PA)-induced generation of reactive oxygen species (ROS) and further explored the underlying molecular mechanisms. We found that resveratrol significantly reduced the PA-induced endothelial ROS levels in human aortic endothelial cells. Resveratrol also induced endothelial cell autophagy, which mediated the effect of resveratrol on ROS reduction. Resveratrol stimulated autophagy via the AMP-activated protein kinase (AMPK)-mTOR pathway. Taken together, these data suggest that resveratrol prevents PA-induced intracellular ROS by autophagy regulation via the AMPK-mTOR pathway. Thus, the induction of autophagy by resveratrol may provide a novel therapeutic candidate for cardioprotection in .

Keyword: metabolic syndrome

Apolipoprotein E isoforms 3/3 and 3/4 differentially interact with circulating stearic, , and oleic fatty acids and lipid levels in Alaskan Natives.

Lifestyle changes in Alaskan Natives have been related to the increase of cardiovascular disease and in the last decades. Variation of the apolipoprotein E (Apo E) genotype may contribute to the diverse response to diet in lipid metabolism and influence the association between fatty acids in plasma and risk factors for cardiovascular disease. The aim of this investigation was to analyze the interaction between Apo E isoforms and plasma fatty acids, influencing phenotypes related to diseases in Alaskan Natives. A sample of 427 adult Siberian Yupik Alaskan Natives was included. Fasting glucose, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, Apo A1, and Apo B plasma concentrations were measured using reference methods. Concentrations of 13 fatty acids in fasting plasma were analyzed by gas chromatography, and Apo E variants were identified. Analyses of covariance were conducted to identify Apo E isoform and fatty main effects and multiplicative interactions. The means for body mass index and age were 26 ± 5.2 and 47 ± 1.5, respectively. Significant main effects were observed for variation in Apo E and different fatty acids influencing Apo B levels, triglycerides, and total cholesterol. Significant interactions were found between Apo E isoform and selected fatty acids influencing total cholesterol, triglycerides, and Apo B concentrations. In summary, Apo E3/3 and 3/4 isoforms had significant interactions with circulating levels of stearic, , oleic fatty acids, and phenotypes of lipid metabolism in Alaskan Natives.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

[The unity of pathogenesis of insulin resistance and non-alcoholic fatty disease of liver. The disorder of fatty acids and triglycerides].

The pathogenesis of non-alcoholic fatty disease of liver (steatosis) is still as unclear as a loss of hepatocytes similar to apoptosis, development of biological reaction of inflammation, its transformation into steatohepatitis with subsequent fibrosis and formation of atrophic cirrhosis. The article suggests that steatosis is developed due to higher concentration of saturated fatty (C 16:0) in food, intensification of its endogenic synthesis from food carbohydrates and glucose and development of insulin resistance. It is displayed in in hormone ability to activate both oxidation in cells of glucose and synthesis of oleic monoene fatty from saturated fatty (C 18:1). The insulin resistance initiates pathologic process on the level of paracrine associations of cells resulting in permanent increase of concentration of non-etherified fatty acids in intercellular medium and intensification of their passive absorption by cells. The phylogenetically ancient mitochondrions will not to oxidize glucose until non-etherified fatty acids are present in cytosol and hence there is an opportunity to oxidize them. To eliminate undesirable action of polar saturated fatty , the cells etherify it by spirit glyceride into triglycerides to deposit in cytosol or to secrete into blood in a form of lipoproteins of very low density. Under insulin resistance, saturated fatty synthesized by hepatocytes from glucose, does not further transform into oleic monoenic fatty . The cells are to etherify endogenic (exogenic) palmnitic saturated fatty into composition of aphysiologic triglycerides (saturated fatty in position sn-2 of spirit glyceride). At that, triglycerides of palmitat-palmitat-oleat and even tripalmitat type are formed. The melting temperature of tripalmitat is 48 degrees C and melting temperature of physiologic trioletat is 13 degrees C. The intracellular lipases factually can\'t hydrolyze triglycerides. So, hepatocytes, overloaded by them, are destroyed in a way similar to apoptosis. The formed corpuscles of apoptosis disorder the biologic function of endoecology and trigger biologic reaction of inflammation. At that, steatosis changes into steato-hepatitis. The prevention of steatosis consists in dramatic restriction of concentration of saturated fatty in food. The treatment effect is targeted to: decreasing the formation of palmitine triglycerides by force of concurrent etherification of saturated fatty not into triglycerides but into phosphatidylcholine (symmetric phospholipids of soya); intensification of oxidation of saturated fatty in peroxisomes (glytazones and fibrates); decrease of insulin resistance (binuanide metformine).

Keyword: metabolic syndrome

Metformin reduces lipid accumulation in macrophages by inhibiting FOXO1-mediated transcription of fatty -binding protein 4.

The accumulation of lipids in macrophages contributes to the development of atherosclerosis. Strategies to reduce lipid accumulation in macrophages may have therapeutic potential for preventing and treating atherosclerosis and cardiovascular complications. The antidiabetic drug metformin has been reported to reduce lipid accumulation in adipocytes. In this study, we examined the effects of metformin on lipid accumulation in macrophages and investigated the mechanisms involved.We observed that metformin significantly reduced (PA)-induced intracellular lipid accumulation in macrophages. Metformin promoted the expression of carnitine palmitoyltransferase I (CPT-1), while reduced the expression of fatty -binding protein 4 (FABP4) which was involved in PA-induced lipid accumulation. Quantitative real-time PCR showed that metformin regulates FABP4 expression at the transcriptional level. We identified forkhead transcription factor FOXO1 as a positive regulator of FABP4 expression. Inhibiting FOXO1 expression with FOXO1 siRNA significantly reduced basal and PA-induced FABP4 expression. Overexpression of wild-type FOXO1 and constitutively active FOXO1 significantly increased FABP4 expression, whereas dominant negative FOXO1 dramatically decreased FABP4 expression. Metformin reduced FABP4 expression by promoting FOXO1 nuclear exclusion and subsequently inhibiting its activity.Taken together, these results suggest that metformin reduces lipid accumulation in macrophages by repressing FOXO1-mediated FABP4 transcription. Thus, metformin may have a protective effect against lipid accumulation in macrophages and may serve as a therapeutic agent for preventing and treating atherosclerosis in .Copyright 2010 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

PINK1-Parkin-Mediated Mitophagy Protects Mitochondrial Integrity and Prevents Stress-Induced Endothelial Injury.

Mitochondrial injury and dysfunction, a significant feature in , triggers endothelial cell dysfunction and cell death. Increasing evidence suggests that mitophagy, a process of autophagic turnover of damaged mitochondria, maintains mitochondrial integrity. PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase 1) and Parkin signaling is a key pathway in mitophagy control. In this study, we examined whether this pathway could protect mitochondria under stress. We found that (PA) induced significant mitophagy and activated PINK1 and Parkin in endothelial cells. Knocking down PINK1 or Parkin reduced mitophagy, leading to impaired clearance of damaged mitochondria and intracellular accumulation of mitochondrial fragments. Furthermore, PINK1 and Parkin prevented PA-induced mitochondrial dysfunction, ROS production and apoptosis. Finally, we show that PINK1 and Parkin were up-regulated in vascular wall of obese mice and diabetic mice. Our study demonstrates that PINK1-Parkin pathway is activated in response to stress. Through induction of mitophagy, this pathway protects mitochondrial integrity and prevents stress-induced endothelial injury.

Keyword: metabolic syndrome

Castration modifies aortic vasoreactivity and serum fatty acids in a sucrose-fed rat model of .

Levels of testosterone and estradiol influence the incidence of cardiovascular diseases: generally, estrogens in females are protective before menopause; coronaropathies, hypertension, and dyslipidemias in normal men are more frequent at comparable ages. We investigated the modulation by castration of in vitro vasoreactivity, serum lipid content, and systolic blood pressure (SBP) in rats with sucrose-induced . The main characteristics of the rat model are: hypertriglyceridemia, moderately high blood pressure, intra-abdominal accumulation of adipose tissue, hyperinsulinemia, nephropathy, increased oxidative stress, and altered vasoreactivity. Male weanling rats received 30% sucrose solution for 16 weeks (; MS), controls (C) had plain water; both had commercial rodent chow. They were subdivided into five groups with two subgroups each: Group 1, intact C and MS rats, Groups 2-5, C and MS rats castrated for periods of 16, 12, 8, and 4 weeks. At the end of the study period, systolic blood pressure was measured, and blood and aortas were obtained for fatty determination and vasoreactivity assays, respectively. After 16 weeks\' sucrose treatment MS aortas showed hypercontractility and decreased vasodilation. and palmitoleic acids were increased in MS versus C. Arachidonic levels in MS were lower than in intact or castrated C. Long-term castration of 16 weeks normalized the levels of and oleic acids. With the shorter periods of castration, contractility increased and relaxation decreased in C and MS, but it was more significant in C. Regarding fatty composition, long-term castration increased polyunsaturated (arachidonic and eicosapentaenoic) fatty acids. The shorter periods did not modify the fatty profile in either C or MS. altered SBP, aortic reactivity, and levels of fatty acids; castration of long duration normalized them in some cases.

Keyword: metabolic syndrome

Simvastatin inhibits LPS-induced alveolar bone loss during .

Studies in recent years have shown a positive relationship between (MS) and periodontal disease (PD). Given that patients with MS take statins to reduce cholesterol, and statins also have anti-inflammatory effects, it is important to determine if statin intake hinders the progression of MS-associated PD. In this study, PD was induced in Zucker fat rats (ZFRs), an animal model for MS, and in control lean rats by periodontal injection of Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS), while simvastatin was given to some of the rats via gavage. After 4 wk of treatment, alveolar bone loss was determined by micro-computed tomography. To explore the underlying mechanisms, we determined the effect of simvastatin on tissue inflammation and the expression of molecules involved in osteoclastogenesis. Results showed that while bone loss was increased by LPS in both ZFRs and the control lean rats, it was significantly more in the former than the latter. Simvastatin effectively alleviated bone loss in both ZFRs and the control rats. Results also showed that LPS stimulated leukocyte tissue infiltration and expression of molecules for osteoclastogenesis, but simvastatin significantly modulated the stimulation. This study demonstrated that simvastatin inhibited LPS-induced alveolar bone loss and periodontal tissue inflammation in rats with MS.

Keyword: metabolic syndrome

Dietary fatty acids modulate adipocyte TNFa production via regulation of its DNA promoter methylation levels.

The factors regulating TNF alpha (TNFa) levels could be considered therapeutic targets against development. DNA methylation is a potent regulator of gene expression and may be associated with protein levels. In this study we investigate whether the effect of dietary fatty acids on TNFa released from adipocytes might be associated with modifications of the TNFa promoter DNA methylation status. A group of rats was assigned to three diets with a different composition of saturated, monounsaturated and polyunsaturated fatty acids. Samples of visceral adipose tissues were taken for adipocyte isolation, in which released TNFa levels were measured, and for methylation and expression studies. In addition, 3 T3-L1 cells were treated with , oleic and linoleic acids, with and without 5-Azacitydine (5-AZA). After treatments, cells and supernatants were included in the same analyses as rat samples. TNFa promoter methylation levels, gene expression and secretion were different according to the diets and fatty treatments associated with them. Cells treated with 5-AZA displayed higher TNFa levels than in the absence of 5-AZA, without differences between fatty acids. According to our results, dietary fatty regulation of adipocyte TNFa levels may be mediated by epigenetic modifications of the TNFa promoter DNA methylation levels.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

[Breast milk: biological constituents for health and well-being in infancy].

Many factors are involved in infants\' health; one of the most important of them may be the kind of early feeding. Recent evidences suggest that breastfeeding, in addition to its well-established beneficial effects during lactation period, provides also beneficial long-term effects, like the protection against infectious and immune-related diseases, a better cognitive development, a decreased risk of and of obesity. It has been reported that the early feeding mode affects growth and body composition and it could be considered a critical factor for development. Human milk is a source of different nutrients and bioactive factors, especially hormones and growth factors like leptin, ghrelin, insulin, insulin-like growth factor (IGF-I) playing a role in food intake regulation, metabolism and body composition. In particular breast milk leptin may provide a physiological explanation for a number of advantages seen in reaching proper growth and energy balance in breast-fed infants compared with formula fed ones. Etiopathogenesis and therapeutic approach in common minor gastrointestinal diseases in infants are important subject of study for pediatricians. Colic, constipation and regurgitation can be considered feeding problems and they might benefit from dietary treatment. Regarding infantile colic, dietary modifications seem to be more suitable than pharmacological treatment in resolving symptoms; also prebiotics and probiotics are useful for this aim. The occurence of constipation is related to the kind of feeding and it is lower in breastfed infants. Moreover formulas with probiotics and beta- could promote a regression of symptoms. A dietary approach may be useful also in regurgitation. Anyway we have to remember that breastfeeding require a supplementation of vitamin D and K for some months and a correct weaning program is needed from the 5th-6th months of life to prevent iron deficiency.

Keyword: metabolic syndrome

[The oleic triglycerides of palm oil and triglycerides of creamy fat. The reaction of palmitoylation, potassium and magnesium palmitate, absorption of fatty acids by enterocytes and microbiota of large intestine].

The decreasing of content of animal, milk fat (butter) by means of its substitution with vegetable, oleic, palmy oil in food of adults optimal by its quantity is physically chemically and biologically substantiated. In oleic palmy oil higher content of oleic mono unsaturated fatty and oleic triglycerides than in creamy fat is established. The biologic availability of unsaturated in the form of free fatty is decreased at its absorption by enterocytes of small intestines is detected. There are no transforms of mono unsaturated acids in palmy oil in contrast with hydrogenated margarines. In palmy, oleic oil there is not enough of short-chained fatty acids (C4-C6) and it has no taste quality and it has low level of unsaturated fatty acids and factually it is lacking of ω-6 polyunsaturated fatty acids. However, it is compensated in case of availability offish and sea products in food. If adults, especially older ones, will refuse to consume creamy fat and decrease intake of products with high content of unsaturated fatty and triglycerides (beef, sour cream, fatty cheeses) it\'ll positively impact their health. The refusal from these products is a real step in prevention of pandemic (atherosclerosis and atheromatosis, , resistance to insulin, obesity). There are still large number of people who at optimal amount of food retain in vivo increased amount of exogenous, endogenously synthesized from glucose unsaturated fatty in the form of unesterified fatty acids ( of resistance to insulin) and increased content of triglycerides.

Keyword: metabolic syndrome

Protective effects of a compound herbal extract (Tong Xin Luo) on free fatty induced endothelial injury: implications of antioxidant system.

Tong-Xin-Luo (TXL) - a mixture of herbal extracts, has been used in Chinese medicine with established therapeutic efficacy in patients with coronary artery disease.We investigated the protective role of TXL extracts on endothelial cells injured by a known risk factor - (PA), which is elevated in and associated with cardiovascular complications. Human aortic endothelial cells (HAECs) were preconditioned with TXL extracts before exposed to PA for 24 hours.We found that PA (0.5 mM) exposure induced 73% apoptosis in endothelial cells. However, when HAECs were preconditioned with ethanol extracted TXL (100 microg/ml), PA induced only 7% of the endothelial cells into apoptosis. Using antibody-based protein microarray, we found that TXL attenuated PA-induced activation of p38-MAPK stress pathway. To investigate the mechanisms involved in TXL\'s protective effects, we found that TXL reduced PA-induced intracellular oxidative stress. Through AMPK pathway, TXL restored the intracellular antioxidant system, which was depressed by the PA treatment, with an increased expression of thioredoxin and a decreased expression of the thioredoxin interacting protein.In summary, our study demonstrates that TXL protects endothelial cells from PA-induced injury. This protection is likely mediated by boosting intracellular antioxidant capacity through AMPK pathway, which may account for the therapeutic efficacy in TXL-mediated cardiovascular protection.

Keyword: metabolic syndrome

[The and oleic modes of metabolism of fatty acids. The exogenous of resistance to insulin under disorder of biologic function of nutrition (trophology)].

The hyperglycemia and insulin are two phylogenetically different humoral regulators of metabolism in vivo. The development of hyperglycemia occurred billions years hitherto under implementation of nutrition function. The insulin was formed in the process of development of biologic function of locomotion. The of resistance to insulin consists in the derangement of humoral regulation of metabolism of fatty acids and glucose at the phylogenetically different levels in vivo both in paracrine cells cenosis and at the level of organism. The exogenous and endogenic syndromes of resistance to insulin are distinguished. The exogenous resistance to insulin is formed under physiologic function of insulin system when hormone effect is prevented by derangement of biologic function of trophology (nutrition)--the formation of such palmitinic mode of metabolism of fatty acids as substrates for oxidation in mitochondria. The endogenic of resistance to insulin consists in discrepancy of regulation of biologic functions at the level of organism under realization of locomotion function and at the level of paracrine cells cenosis under realization of biologic function of adaptation, endoecology (support of "cleanness" of intracellular medium) and its biologic reaction of inflammation, homeostasis function. The of resistance to insulin is energetic issue in vivo. Primarily, insulin regulates metabolism of fatty acids and only secondly transformations of glucose. In case ofpalmitinic mode of metabolism offatty acids in the enzymes with the same parameters are involved in biologic reactions. The palmitinic triglycerides are not optimal due to aphysiological slow biochemical and physico-chemical reactions.

Keyword: metabolic syndrome

Global profiling of infection by an oncogenic virus: KSHV induces and requires lipogenesis for survival of latent infection.

Like cancer cells, virally infected cells have dramatically altered requirements. We analyzed global changes induced by latent infection with an oncogenic virus, Kaposi\'s Sarcoma-associated herpesvirus (KSHV). KSHV is the etiologic agent of Kaposi\'s Sarcoma (KS), the most common tumor of AIDS patients. Approximately one-third of the nearly 200 measured metabolites were altered following latent infection of endothelial cells by KSHV, including many metabolites of anabolic pathways common to most cancer cells. KSHV induced pathways that are commonly altered in cancer cells including glycolysis, the pentose phosphate pathway, amino production and fatty synthesis. Interestingly, over half of the detectable long chain fatty acids detected in our screen were significantly increased by latent KSHV infection. KSHV infection leads to the elevation of metabolites involved in the synthesis of fatty acids, not degradation from phospholipids, and leads to increased lipid droplet organelle formation in the infected cells. Fatty synthesis is required for the survival of latently infected endothelial cells, as inhibition of key enzymes in this pathway led to apoptosis of infected cells. Addition of to latently infected cells treated with a fatty synthesis inhibitor protected the cells from death indicating that the products of this pathway are essential. Our metabolomic analysis of KSHV-infected cells provides insight as to how oncogenic viruses can induce alterations common to cancer cells. Furthermore, this analysis raises the possibility that pathways may provide novel therapeutic targets for the inhibition of latent KSHV infection and ultimately KS tumors.

Keyword: metabolic syndrome

Dietary fat, fatty composition in plasma and the .

The , a cluster of disorders often including abdominal obesity, is associated with a high risk of cardiovascular disease and premature death. Insulin resistance is a key feature of the . Observational studies have indicated that the type of fat in the diet may be related to the development of insulin resistance and the , also independent of possible effects on body weight. Dietary surveys are often imprecise. One way to monitor the type of fat in the diet is to record the fatty composition in plasma. This review summarizes recent data on the relationships between fatty composition in plasma and insulin resistance, diabetes and other disorders related to the .Insulin resistance and insulin resistant states are often associated with the fatty pattern in plasma, characterized by an increased proportion of (16 : 0) and a low proportion of linoleic (18 : 2 n-6) acids, with a distribution of other fatty acids indicating an increased activity of delta-9 and delta-6 desaturase. This shows that there may be a causal relationship between the type of fat in the diet and insulin action, an assumption supported by recent dietary intervention studies.In a public health perspective these results, from both observational and intervention studies, underline the importance of fat quality in the diet for the development of a number of prevalent diseases. Taken together with several earlier studies and recent epidemiological findings, they give strong support to present dietary guidelines.

Keyword: metabolic syndrome

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: metabolic syndrome

Dietary interventions for : role of modifying dietary fats.

In humans, insulin sensitivity is relatively impaired by diets that are low in oleic (OA), a cis monounsaturated fatty (MUFA), or rich in trans MUFA or (PA), a saturated fatty (FA). Emerging evidence exists that PA, in contrast to OA, causes insulin resistance via stimulation of inflammatory signaling and production of cytosolic lipid compounds (diacylglycerol and ceramide), leading one to presume that dietary or pharmacologic maneuvers that facilitate transport of FA into the mitochondria would be beneficial. However, in some models, insulin resistance is caused by excessive FA transport into the mitochondria, coupled with deficient electron transport and possibly increased reactive oxygen species formation; PA may impair electron transport via effects on gene expression. A research challenge is to determine whether feeding humans diets with markedly different contents of PA and OA would alter insulin sensitivity and/or critical biochemical mechanisms impacting muscle insulin signaling.

Keyword: metabolic syndrome

[Features of fatty- spectrum of triglycerides in patients with insulin resistance and ].

The article presents results of the study of fatty- spectrum of triglycerides in insulin resistance patients with . An analysis was done on fatty- spectrum of triglycerides in healthy men and women, difference between types of dislipidemia diagnosed in men and women, including women in regard to their reproductive function. The authors found increase in level in fatty- spectrum of triglycerides in patients with diabetes mellitus type 2. Conclusion was made on correction of patient diet at the expense of ratio of saturated, nonsaturated and essential polyene acids: decrease in quantity of and increase in oleic for primary and secondary prevention atherosclerosis.

Keyword: metabolic syndrome

[The individual fatty acids in blood plasma, erythrocytes and lipoproteins. The comparison of tests results of patients with ischemic heart disease and volunteers].

According to the generally accepted theory, the atherosclerosis is a kind of disorder of metabolism of lipids which chemically are the ethers of fatty lipids with spirits. Hence, the atherosclerosis is fatty acids pathology. In conformity with the biologic classification, among fatty acids it is functionally valid to distinguish saturated fatty acids without double bonds; monoenic fatty acids with one double bond; unsaturated fatty acids with two or three double bonds and polyenic fatty acids with four of six double bonds in chain. The saturated and monenic fatty acids are the substrates for cells to groundwork energy, ATP The unsaturated fatty acids in vivo are needed to form membranes. The polyenic fatty acids are essential since they are precursors of cell synthesis of humoral regulators--eicosanoids (prostanoids and leukotrienes). To clarify the pathogenesis of the " pandemics" most prevalent in human population, the quantitative determination of individual fatty acids in blood plasma and erythrocytes using gas chromatography technique is needed. It is necessary to evaluate the content of medium chain fatty acids; and stearic saturated fatty acids; oleic monoenic fatty and its transforms--linoleic, linolenic and dihomo-gamma-linolenic unsaturated fatty acids; essential polyenic omega-6 arachidonic, omega-3 eicosapentaenoic and docosahexaenoic fatty acids. The higher is in food the content of saturated fatty , palmitoleic and trans-vaccenic monoenic fatty acids, the more is in patient diet of beef meat and products of fat cow\'s milk. The higher is ratio of /oleic fatty acids the lower is the risk of formation of atheromatosis of arteries intima and development of ischemic heart disease and vice versa. The decrease of ratio of omega-3/omega-6 essential polyenic fatty acids is undesirable in prognostic sense. The metabolism of these acids differs and functional activity of omega-3 eicosanoid type 3 is higher In case of deficiency of omega-3 and omega-6 polyenic fatty acids in cells eicosanoids are synthesized from unsaturated dihomogamma-linolenic fatty and their influence turns out to be aphysiologic. This condition is a pathogenic foundation of atherosclerosis. There is a diagnostic reason to detect fatty acids in case of diabetes mellitus, obesity, and partially arterial hypertension.

Keyword: metabolic syndrome

Associations of erythrocyte fatty acids in the de novo lipogenesis pathway with risk of in a cohort study of middle-aged and older Chinese.

Experimental studies suggest that elevated de novo lipogenesis (DNL) might be involved in the pathogenesis of disorders. Few prospective studies have been conducted, especially among populations with a high carbohydrate intake, to determine whether DNL fatty acids are associated with the risk of the (MetS).We aimed to investigate associations of erythrocyte fatty acids in the DNL pathway-including myristic (14:0), (16:0), palmitoleic (16:1n-7), hexadecenoic (16:1n-9), stearic (18:0), vaccenic (18:1n-7), and oleic (18:1n-9)-with the risk of MetS in a Chinese population with an average carbohydrate intake of >60% of energy.A total of 1176 free-living Chinese men and women aged 50-70 y from Beijing and Shanghai were included in our analysis, giving rise to 412 incident MetS cases during 6 y of follow-up. Erythrocyte fatty acids and traits were measured in these participants.Erythrocyte fatty acids in the DNL pathway were correlated with a high ratio of carbohydrate-to-fat intake, less favorable lipid profiles, and elevated liver enzymes at baseline. In comparison with the lowest quartile, RRs (95% CIs) of MetS in the highest quartile were 1.30 (1.04, 1.62; P-trend = 0.007) for 16:1n-7, 1.48 (1.17, 1.86; P-trend < 0.001) for 16:1n-9, 1.26 (1.01, 1.56; P-trend = 0.06) for 18:1n-7, and 1.51 (1.19, 1.92; P-trend < 0.001) for 18:1n-9 after multivariate adjustment for lifestyle factors and body mass index. Moreover, 16:0 and 16:1n-7 were associated with an elevated risk of diabetes.Our findings suggest that fatty acids in the DNL pathway are independently associated with an elevated risk of disorders.

Keyword: metabolic syndrome

CD36 is upregulated in mice with periodontitis and and involved in macrophage gene upregulation by palmitate.

We reported that high-fat diet (HFD)-induced (MetS) exacerbates lipopolysaccharide (LPS)-stimulated periodontitis and palmitate, the major saturated fatty in the HFD, amplified LPS-stimulated gene expression in\xa0vitro. As CD36 is a major receptor for fatty acids, we investigated periodontal CD36 expression in mice with periodontitis and MetS, and the role of CD36 in inflammatory gene expression in macrophages stimulated by palmitate.MetS and periodontitis were induced in mice by HFD and periodontal injection of LPS, respectively. The periodontal CD36 expression and its relationship with alveolar bone loss were studied using immunohistochemistry, real-time PCR, and correlation analysis. The role of CD36 in upregulation of inflammatory mediators by LPS and palmitate in macrophages was assessed using pharmacological inhibitor and small interfering RNA.Periodontal CD36 expression was higher in mice with both MetS and periodontitis than that in mice with periodontitis or MetS alone and was correlated with osteoclastogenesis and alveolar bone loss. In vitro studies showed that CD36 expression in macrophages was upregulated by LPS and palmitate, and targeting CD36 attenuated palmitate-enhanced gene expression.CD36 expression is upregulated in mice with periodontitis and MetS and involved in gene expression in macrophages stimulated by palmitate and LPS.© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: metabolic syndrome

[REFSUM\'S (HEREDOPATHIA ATACTICA POLYNEURITIFORMIS): CONGENITAL DISORDER OF LIPID METABOLISM WITH STORAGE OF 3,7,11,15-TETRAMETHYL-HEXADECANOIC ].

Keyword: metabolic syndrome

Plasma profiling on postoperative colorectal cancer patients with different traditional Chinese medicine syndromes.

This study aims to investigate the profiles of postoperative colorectal cancer (PCRC) patients with different traditional Chinese medicine (TCM) syndromes and to discuss the mechanism under PCRC progression and TCM classification.Fifty healthy controls (HC) and 70 PCRC patients, including 10 Dampness and heat (DHS), 33 Spleen deficiency (SDS), 19 Liver and kidney Yin deficiency (LKYDS) and 8 with non-TCM (NS) were enrolled. Plasma profiles were detected by Gas chromatography-mass spectrometry (GC-MS) and analyzed by principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA). Furthermore, pathway enrichment was analyzed based on KEGG and DAVID databases and network was constructed via metaboanalyst and cytoscape.The top-3 metabolites with higher abundance in PCRC compared with HC were terephthalic (165.417-fold), ornithine (24.484-fold) and aminomalonic (21.346-fold). And the cholesterol (0.588-fold) level was decreased in PCRC. l-Alanine, 1, 2-ethanediamine, urea, glycerol, glycine, aminomalonic , creatinine and were specifically altered in the DHS, while d-tryptophan was exclusively changed in SDS, and l-proline, 1, 2, 3-propanetricarboxylic , d-galactose and 2-indolecarboxylic acids in LKYDS.The plasma profiles were perturbed in PCRC patients. Increased levels of terephthalic might indicate high risk of relapse and elevated ornithine may contribute to the post-operational recovery or may raise the susceptibility to PCRC recurrence. The profiles of DHS, SDS, LKYDS and NS were almost separately clustered, indicating the possibility of explaining TCM syndromes classification using metabolomics. Furthermore, creatinine and aminomalonic alternation might correlate with the formation of DHS, while d-tryptophan may associate with SDS and d-galactose and 1, 2, 3-propanetricarboxylic may relate to LKYDS. As numbers of patients in each TCM are small, further study is needed to verify those results.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: metabolic syndrome

Long-term exposition to a high fat diet favors the appearance of β-amyloid depositions in the brain of C57BL/6J mice. A potential model of sporadic Alzheimer\'s disease.

The sporadic and late-onset form of Alzheimer\'s disease (AD) constitutes the most common form of dementia. This non-familiar form could be a consequence of , characterized by obesity and the development of a brain-specific insulin resistance known as type III diabetes. This work demonstrates the development of a significant AD-like neuropathology due to these alterations.C57BL/6J mice strain were divided into two groups, one fed with a diet rich in (high-fat diet, HFD) since their weaning until 16 months of age, and another group used as a control with a regular diet. The analyses were carried out in the dentate gyrus area of the hippocampus using a Thioflavin-S stain and immunofluorescence assays.The most significant finding of the present research was that HFD induced the deposition of the βA peptide. Moreover, the diet also caused alterations in different cell processes, such as increased inflammatory reactions that lead to a decrease in the neuronal precursor cells. In addition, the results show that there were also dysregulations in normal autophagy and apoptosis, mechanisms related to βA formation.The present findings confirm that HFD favors the formation of βA depositions in the brain, a key feature of AD, supporting the hypothesis of sporadic AD.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: metabolic syndrome

Metformin plays a dual role in MIN6 pancreatic β cell function through AMPK-dependent autophagy.

Metformin improves insulin sensitivity in insulin sensitive tissues such as liver, muscle and fat. However, the functional roles and the underlying mechanism of metformin action in pancreatic β cells remain elusive. Here we show that, under normal growth condition, metformin suppresses MIN6 β cell proliferation and promotes apoptosis via an AMPK-dependent and autophagy-mediated mechanism. On the other hand, metformin protects MIN6 cells against (PA)-induced apoptosis. Our findings indicate that metformin plays a dual role in β cell survival and overdose of this anti-diabetic drug itself may lead to potential β cell toxicity.

Keyword: metabolic syndrome

Nrf2 affects the efficiency of mitochondrial fatty oxidation.

Transcription factor Nrf2 (NF-E2 p45-related factor 2) regulates the cellular redox homoeostasis and cytoprotective responses, allowing adaptation and survival under conditions of stress. The significance of Nrf2\xa0in intermediary metabolism is also beginning to be recognized. Thus this transcription factor negatively affects fatty synthesis. However, the effect of Nrf2 on fatty oxidation is currently unknown. In the present paper, we report that the mitochondrial oxidation of long-chain () and short-chain (hexanoic) fatty acids is depressed in the absence of Nrf2 and accelerated when Nrf2 is constitutively active. Addition of fatty acids stimulates respiration in heart and liver mitochondria isolated from wild-type mice. This effect is significantly weaker when Nrf2 is deleted, whereas it is stronger when Nrf2 activity is constitutively high. In the absence of glucose, addition of fatty acids differentially affects the production of ATP in mouse embryonic fibroblasts from wild-type, Nrf2-knockout and Keap1 (Kelch-like ECH-associated protein 1)-knockout mice. In acute tissue slices, the rate of regeneration of FADH2 is reduced when Nrf2 is absent. This role of Nrf2 on fatty oxidation has implications for chronic disease conditions including cancer, and neurodegeneration.

Keyword: metabolic syndrome

Effects of 1-year intervention with a Mediterranean diet on plasma fatty composition and in a population at high cardiovascular risk.

(MetS) has become an important public concern due to its increasing prevalence. An altered fatty composition has been associated with MetS, but the Mediterranean diet has been shown to have a protective effect. The aim of the present study was to analyze the influence of a Mediterranean dietary pattern, as assessed by the biomarkers of food supplied, on the plasma fatty composition and its relation with MetS after 1 year of intervention.A total of 424 subjects were randomly selected from the PREDIMED randomized dietary trial after completing a 1-year intervention program. Participants aged 55 to 80 years and at high risk of cardiovascular disease were randomly assigned to three dietary interventions: Mediterranean diet supplemented with virgin olive oil or nuts, or a low-fat diet.After 1 year of intervention participants in the virgin olive oil group showed significantly increased plasma concentrations of and oleic acids, but reduced proportions of margaric, stearic, and linoleic acids. In turn, subjects in the nut group showed significantly increased levels of , linoleic, and α-linolenic acids, but reduced proportions of myristic, margaric, palmitoleic, and dihommo-γ-linoleic acids. Increases in the biomarkers of foods supplied to the Mediterranean diet groups, i.e., oleic and α-linolenic acids, were beneficially associated with the incidence, reversion and prevalence of MetS. No weight changes were observed among participants.The nut and olive oil diets induced a fatty composition that has been shown to be beneficial in the face of MetS. Therefore, a Mediterranean diet rich in fats of vegetable origin may be a useful tool for the management of MetS without the need for concerns over weight gain due to its high fat content.Controlled-Trials.com ISRCTN35739639.

Keyword: metabolic syndrome

Renal amino , fat and glucose metabolism in type 1 diabetic and non-diabetic humans: effects of acute insulin withdrawal.

The aim of this study was to test the hypothesis that type 1 diabetes alters renal amino , glucose and fatty metabolism.We studied five C-peptide-negative, type 1 diabetic subjects during insulin replacement (glucose 5.6 mmol/l) and insulin deprivation (glucose 15.5 mmol/l) and compared them with six non-diabetic subjects. Leucine, phenylalanine, tyrosine, glucose and palmitate tracers were infused after an overnight fast and samples were obtained from the renal vein, femoral vein and femoral artery.Insulin deprivation significantly increased whole-body fluxes (20-25%) of phenylalanine, tyrosine and leucine, and leucine oxidation (50%). Kidney contributed 5-10% to the whole-body leucine and phenylalanine flux. A net uptake of phenylalanine, conversion of phenylalanine to tyrosine (5 micromol/min) and net release of tyrosine (approximately 5 micromol/min) occurred across the kidney. Whole-body (three-fold) and leg (two-fold) leucine transamination increased but amino metabolism in the kidney did not alter with diabetes or insulin deprivation. Insulin deprivation doubled endogenous glucose production, renal glucose production was unaltered by insulin deprivation and diabetes (ranging between 100 and 140 micromol/min). Renal palmitate exchange was unaltered by insulin deprivation.In conclusion, kidney post-absorptively accounts for 5-10% of whole-body protein turnover, 15-20% of leucine transamination and 10-15% of endogenous glucose production, and actively converts phenylalanine to tyrosine. During insulin deprivation, leg becomes a major site for leucine transamination but insulin deprivation does not affect renal phenylalanine, leucine, palmitate or glucose metabolism. Despite its key role, insulin deprivation in type 1 diabetic patients does not alter many of these functions.

Keyword: metabolic syndrome

UHPLC/Q-TOFMS-based plasma metabolomics of polycystic ovary patients with and without insulin resistance.

Polycystic ovary (PCOS), characterized with menstrual irregularities, hyperandrogenism and ovulatory abnormalities, is usually companied with insulin resistance (IR) and accounts for one of the most prevalent reproductive dysfunction of premenopausal women. Despite accumulating investigations, diagnostic standards of this pathological condition remain obscure. The aim of present study is to characterize the plasma characteristics of PCOS patients with and without IR, and subsequently identify the potential biomarkers for the diagnosis of PCOS and its IR complication. A total of 59 plasma samples from eligible healthy controls (CON, n=19), PCOS patients without IR (non-IR PCOS, n=19) and PCOS patients with IR (IR PCOS, n=21) were profiled by an ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOFMS) followed by multivariate statistical analysis. Compared to the healthy controls, significant decrease in the levels of phosphocholines (PCs) and lyso PC (18:2), and increase in trilauric glyceride level were observed in the plasma of IR PCOS. Meanwhile, the significant increase in the levels of saturated fatty acids ( and stearic ) and decanoylcarnitine, and decrease in PC (36:2) and PS (36:0) were found in non-IR PCOS patients. Trilauric glyceride and decanoylcarnitine were identified as the potential biomarkers with the highest sensitivity and specificity for the diagnosis of PCOS patients with and without IR, respectively. Furthermore, based on these alterations of metabolites, MetPA network pathway analysis suggested a profound involvement of the abnormalities of glycerophospholipid, glycerolipid and fatty metabolisms in the pathogenesis of PCOS and IR complications. Collectively, LC-MS-based metabolomics provides a promising strategy for complementary diagnosis of PCOS and its IR complication and offers a new insight to understand their pathogenesis mechanisms.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Nutraceutical Potential of in .

L. is a well-known fruit worldwide, and its highest production occurs in tropical and subtropical regions. The pulp contains vitamins A, C, and E, B complex vitamins, such as pantothenic and folate, and minerals, such as magnesium and potassium, as well as food fibers. Phenolic compounds, such as benzyl isothiocyanate, glucosinolates, tocopherols (α and δ), β-cryptoxanthin, β-carotene and carotenoids, are found in the seeds. The oil extracted from the seed principally presents oleic fatty followed by , linoleic and stearic acids, whereas the leaves have high contents of food fibers and polyphenolic compounds, flavonoids, saponins, pro-anthocyanins, tocopherol, and benzyl isothiocyanate. Studies demonstrated that the nutrients present in its composition have beneficial effects on the cardiovascular system, protecting it against cardiovascular illnesses and preventing harm caused by free radicals. It has also been reported that it aids in the treatment of diabetes mellitus and in the reduction of cholesterol levels. Thus, both the pulp and the other parts of the plant (leaves and seeds) present antioxidant, anti-hypertensive, hypoglycemic, and hypolipidemic actions, which, in turn, can contribute to the prevention and treatment of obesity and associated disorders.

Keyword: metabolic syndrome

12/15-lipoxygenase products induce inflammation and impair insulin signaling in 3T3-L1 adipocytes.

Inflammation and insulin resistance associated with visceral obesity are important risk factors for the development of type 2 diabetes, atherosclerosis, and the . The 12/15-lipoxygenase (12/15-LO) enzyme has been linked to inflammatory changes in blood vessels that precede the development of atherosclerosis. The expression and role of 12/15-LO in adipocytes have not been evaluated. We found that 12/15-LO mRNA was dramatically upregulated in white epididymal adipocytes of high-fat fed mice. 12/15-LO was poorly expressed in 3T3-L1 fibroblasts and was upregulated during differentiation into adipocytes. Interestingly, the saturated fatty palmitate, a major component of high fat diets, augmented expression of 12/15-LO in vitro. When 3T3-L1 adipocytes were treated with the 12/15-LO products, 12-hydroxyeicosatetranoic (12(S)-HETE) and 12-hydroperoxyeicosatetraenoic (12(S)-HPETE), expression of proinflammatory cytokine genes, including tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein 1 (MCP-1), interleukin 6 (IL-6), and IL-12p40, was upregulated whereas anti-inflammatory adiponectin gene expression was downregulated. 12/15-LO products also augmented c-Jun N-terminal kinase 1 (JNK-1) phosphorylation, a known negative regulator of insulin signaling. Consistent with impaired insulin signaling, we found that insulin-stimulated 3T3-L1 adipocytes exhibited decreased IRS-1(Tyr) phosphorylation, increased IRS-1(Ser) phosphorylation, and impaired Akt phosphorylation when treated with 12/15-LO product. Taken together, our data suggest that 12/15-LO products create a proinflammatory state and impair insulin signaling in 3T3-L1 adipocytes. Because 12/15-LO expression is upregulated in visceral adipocytes by high-fat feeding in vivo and also by addition of in vitro, we propose that 12/15-LO plays a role in promoting inflammation and insulin resistance associated with obesity.

Keyword: metabolic syndrome

Optimal elevation of β-cell 11β-hydroxysteroid dehydrogenase type 1 is a compensatory mechanism that prevents high-fat diet-induced β-cell failure.

Type 2 diabetes ultimately results from pancreatic β-cell failure. Abnormally elevated intracellular regeneration of glucocorticoids by the enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in fat or liver may underlie pathophysiological aspects of the . Elevated 11β-HSD1 is also found in pancreatic islets of obese/diabetic rodents and is hypothesized to suppress insulin secretion and promote diabetes. To define the direct impact of elevated pancreatic β-cell 11β-HSD1 on insulin secretion, we generated β-cell-specific, 11β-HSD1-overexpressing (MIP-HSD1) mice on a strain background prone to β-cell failure. Unexpectedly, MIP-HSD1(tg/+) mice exhibited a reversal of high fat-induced β-cell failure through augmentation of the number and intrinsic function of small islets in association with induction of heat shock, protein kinase A, and extracellular signal-related kinase and p21 signaling pathways. 11β-HSD1(-/-) mice showed mild β-cell impairment that was offset by improved glucose tolerance. The benefit of higher β-cell 11β-HSD1 exhibited a threshold because homozygous MIP-HSD1(tg/tg) mice and diabetic Lep(db/db) mice with markedly elevated β-cell 11β-HSD1 levels had impaired basal β-cell function. Optimal elevation of β-cell 11β-HSD1 represents a novel biological mechanism supporting compensatory insulin hypersecretion rather than exacerbating disease. These findings have immediate significance for current therapeutic strategies for type 2 diabetes.

Keyword: metabolic syndrome

Histidine supplementation improves insulin resistance through suppressed inflammation in obese women with the : a randomised controlled trial.

Increased inflammation and oxidative stress are associated with insulin resistance (IR) and disorders. Serum histidine levels are lower and are negatively associated with inflammation and oxidative stress in obese women. The objective of this study was to evaluate the efficacy of histidine supplementation on IR, inflammation, oxidative stress and disorders in obese women with the (MetS).A total of 100 obese women aged 33-51 years with BMI ≥ 28 kg/m² and diagnosed with MetS were included following a health examination in the community hospital in this randomised, double-blinded, placebo-controlled trial. Participants were allocated to interventions by an investigator using sequentially numbered sealed envelopes and received 4 g/day histidine (n = 50) or identical placebo (n = 50) for 12 weeks. Participants then attended the same clinic every 2 weeks for scheduled interviews and to count tablets returned. Serum histidine, HOMA-IR, BMI, waist circumference, fat mass, serum NEFA, and variables connected to inflammation and oxidative stress were measured at baseline and 12 weeks. Participants, examining physicians and investigators assessing the outcomes were blinded to group assignment. In addition, the inflammatory mechanisms of histidine were also explored in adipocytes.At 12 weeks, a total of 92 participants completed this trail. Compared with the placebo group (n = 47), histidine supplementation significantly decreased HOMA-IR (-1.09 [95% CI -1.49, -0.68]), BMI (-0.86 kg/m² [95% CI -1.55, -0.17]), waist circumference (-2.86 cm [95% CI -3.86, -1.86]), fat mass (-2.71 kg [95% CI -3.69, -1.73]), serum NEFA (-173.26 μmol/l [95% CI -208.57, -137.94]), serum inflammatory cytokines (TNF-α, -3.96 pg/ml [95% CI -5.29, -2.62]; IL-6, -2.15 pg/ml [95% CI -2.52, -1.78]), oxidative stress (superoxide dismutase, 17.84 U/ml [95% CI 15.03, 20.65]; glutathione peroxidase, 13.71 nmol/ml [95% CI 9.65, 17.78]) and increased serum histidine and adiponectin by 18.23 μmol/l [95% CI 11.74, 24.71] and 2.02 ng/ml [95% CI 0.60, 3.44] in histidine supplementation group (n = 45), respectively. There were significant correlations between changes in serum histidine and changes of IR and its risk factors. No side effects were observed during the intervention. In vitro study indicated that histidine suppresses IL6 and TNF mRNA expression and nuclear factor kappa-B (NF-κB) protein production in -induced adipocytes in a dose-dependent manner, and these changes were diminished by an inhibitor of NF-κB.Histidine supplementation could improve IR, reduce BMI, fat mass and NEFA and suppress inflammation and oxidative stress in obese women with MetS; histidine could improve IR through suppressed pro-inflammatory cytokine expression, possibly by the NF-κB pathway, in adipocytes.

Keyword: metabolic syndrome

The Sjögren-Larsson gene encodes a hexadecenal dehydrogenase of the sphingosine 1-phosphate degradation pathway.

Sphingosine 1-phosphate (S1P) functions not only as a bioactive lipid molecule, but also as an important intermediate of the sole sphingolipid-to-glycerolipid pathway. However, the precise reactions and the enzymes involved in this pathway remain unresolved. We report here that yeast HFD1 and the\xa0Sjögren-Larsson (SLS)-causative mammalian gene ALDH3A2 are responsible for conversion of\xa0the S1P degradation product hexadecenal to hexadecenoic . The absence of ALDH3A2 in CHO-K1 mutant cells caused abnormal metabolism of S1P/hexadecenal to ether-linked glycerolipids. Moreover, we demonstrate that yeast Faa1 and Faa4 and mammalian ACSL family members are acyl-CoA synthetases involved in the sphingolipid-to-glycerolipid pathway and that hexadecenoic accumulates in Δfaa1 Δfaa4 mutant cells.\xa0These results unveil the entire S1P pathway: S1P is metabolized to glycerolipids via hexadecenal, hexadecenoic , hexadecenoyl-CoA, and palmitoyl-CoA. From our results we propose a possibility that accumulation of the S1P metabolite hexadecenal contributes to the pathogenesis of SLS.Copyright © 2012 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

[The composition of nonesterified fatty acids in patients with ].

The blood composition of non-etherized fatty acids (NEFA) was studied in 22 patients with (MS) and 11 healthy individuals. The qualitative NEFA composition presented by 31 components of individual fatty acids was analyzed, by taking into account of glucose-insulin homeostatic changes in MS patients: those without insulin resistance (IR) (Group 1) and those with diagnosed IR (Group 2). MS patients with normal insulinemia were ascertained to have lower levels of lauric, myristic, , C24:0, C16:0i acids. With a decrease in the relative quantity of saturated NEFA, the levels of polyunsaturated fatty acids (FA) increased. The proportions of linoleic (C18:2 omega 6) and linolenic (C18:3 omega 3) acids doubled (p < 0.01), arachidonic (C20:4 omega 6) was observed to tend to rise. The cumulative FA index sigma omega 6 increased twofold. In Group 1, the integrated index of changes in the FA series (unsaturation index) was 41% higher than that in the control group (p < 0.05). In Group 2, the vector of changes in the relative quantity of NEFA was similar, but impairments were less marked than that in Group 1. The findings suggest that the development of insulin resistance is preceded by impaired blood cell transfer and absorption of NEFA.

Keyword: metabolic syndrome

8-Hydroxyeicosapentaenoic Decreases Plasma and Hepatic Triglycerides via Activation of Peroxisome Proliferator-Activated Receptor Alpha in High-Fat Diet-Induced Obese Mice.

PPARs regulate the expression of genes involved in lipid homeostasis. PPARs serve as molecular sensors of fatty acids, and their activation can act against obesity and syndromes. 8-Hydroxyeicosapentaenoic (8-HEPE) acts as a PPAR ligand and has higher activity than EPA. However, to date, the PPAR ligand activity of 8-HEPE has only been demonstrated in vitro. Here, we investigated its ligand activity in vivo by examining the effect of 8-HEPE treatment on high fat diet-induced obesity in mice. After the 4-week treatment period, the levels of plasma and hepatic triglycerides in the 8-HEPE-fed mice were significantly lower than those in the HFD-fed mice. The expression of genes regulated by PPARα was significantly increased in 8-HEPE-fed mice compared to those that received only HFD. Additionally, the level of hepatic in 8-HEPE-fed mice was significantly lower than in HFD-fed mice. These results suggested that intake of 8-HEPE induced PPARα activation and increased catabolism of lipids in the liver. We found no significant differences between EPA-fed mice and HFD-fed mice. We demonstrated that 8-HEPE has a larger positive effect on than EPA and that 8-HEPE acts by inducing PPARα activation in the liver.

Keyword: metabolic syndrome

PTEN phosphorylation and nuclear export mediate free fatty -induced oxidative stress.

Oxidative stress induced by free fatty acids (FFA) contributes to -associated development of cardiovascular diseases, yet molecular mechanisms remain poorly understood. This study aimed at establishing whether phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and its subcellular location play a role in FFA-induced endothelial oxidative stress.Exposing human endothelial cells (ECs) with FFA activated mammalian target of rapamycin (mTOR)/S6K pathway, and upon activation, S6K directly phosphorylated PTEN at S380. Phosphorylation of PTEN increased its interaction with its deubiquitinase USP7 in the nucleus, leading to PTEN deubiquitination and nuclear export. The reduction of PTEN in the nucleus, in turn, decreased p53 acetylation and transcription, reduced the expression of the p53 target gene glutathione peroxidase-1 (GPX1), resulting in reactive oxygen species (ROS) accumulation and endothelial damage. Finally, C57BL/6J mice fed with high-fat atherogenic diet (HFAD) showed PTEN nuclear export, decreased p53 and GPX1 protein expressions, elevated levels of ROS, and significant lesions in aortas. Importantly, inhibition of mTOR or S6K effectively blocked these effects, suggesting that mTOR/S6K pathway mediates HFAD-induced oxidative stress and vascular damage via PTEN/p53/GPX1 inhibition in vivo.Our study demonstrated for the first time that S6K directly phosphorylated PTEN at S380 under high FFA conditions, and this phosphorylation mediated FFA-induced endothelial oxidative stress. Furthermore, we showed that S380 phosphorylation affected PTEN monoubiquitination and nuclear localization, providing the first example of coordinated regulation of PTEN nuclear localization via phosphorylation and ubiquitination.Our studies provide a novel mechanism by which hyperlipidemia causes vascular oxidative damage through the phosphorylation of PTEN, blocking of PTEN nuclear function, and inhibition of p53/GPX1 activity.

Keyword: metabolic syndrome

[THE UNESTERIFIED FATTY ACIDS IN BLOOD PLASMA AND INTERCELLULAR MEDIUM: EFFECT OF INSULIN AND ALBUMIN (THE LECTURE)].

The high content of saturated fatty , triglycerides in food, the large amount of lipoproteins of very low density of the same name in blood, obvious insufficient amount of unesterified fatty acids releasing under lipolysis in blood to meet in vivo requirements in biotransforming energy of ATP are the causes of biological malfunction of homeostasis. As a rule, for every cell in vivo everything is always to be enough. The deficiency of synthesis of ATP by reason of non-optimal substratum for acquirement of ATP by mitochondria is followed by activation also phylogenetically earlier biological function of adaptation, biological reaction of stress. Thus, surplus of unesterified fatty after every food intake forms in vivo biological reaction of "" stress, deficiency of energy by reason of realization by mitochondria in vivo non-optimal exogenous substratum- unesterified fatty , deficiency of acyl- and acetyl-KoA and prognostically formation of potentially ineffective alternative of metabolism of fatty acids. The deficiency of unesterified fatty acids in biological reaction of exotrophy after every intake of food compensates biological reaction of stress, activation of releasing of unesterified fatty acids from visceral fatty cells of gland as it physiologically occurs in biological reaction of endotrophy. At that, adrenalin increases lipolysis in visceral fatty cells of gland and physiologically late insulin can\'t inhibit lipolysis in phylogenetically early visceral fatty cells. Increasing of content of unesterifed fatty acids in blood plasma, as it always occurs in vivo, stops absorption of glucose by cells initiating hyperglycemia, hyperinsulinemia, and of resistance to insulin. The result of such a compensation of biological reaction of exotrophy is biological reaction of endotrophy, condition of "" stress, depletion of function of β-cells of islets with formation of diabetes mellitus type I, deficiency in vivo of insulin synthesis. The biological role of albumin - transfer of fatty acids in intercellular medium inform of unesterifed fatty acids and prevention of formation of pool of free fatty acids effecting aphysiologically.

Keyword: metabolic syndrome

GSK-3β promotes PA-induced apoptosis through changing β-arrestin 2 nucleus location in H9c2 cardiomyocytes.

(PA), a type of saturated fatty acids, induces cardiovascular diseases by causing cardiomyocyte apoptosis with unclear mechanisms. Akt participates in PA-induced cardiomyocyte apoptosis. GSK-3β is a substrate of Akt, we investigated its role in PA-induced apoptosis. We reveal that PA inhibits GSK-3β phosphorylation accompanied by inactivation of Akt in H9c2 cardiomyocytes. We also reveal that inhibition the activity of GSK-3β by its inhibitor LiCl or knockdown by siRNA significantly attenuates PA-induced cardiomyocyte apoptosis, this suggesting that GSK-3β plays a pro-apoptotic role. To detect its downstream factors, we analyzed the roles of JNK, p38 MAPK and β-arrestin 2 (β-Arr2). Here, we report that GSK-3β regulate PA-induced cardiomyocyte apoptosis by affecting the distribution of β-Arr2. PA diminishes the protein level of β-Arr2 and changes its distribution from nucleus to cytoplasm. Either inhibition of β-Arr2 by its siRNA or overexpression of its protein level by transfection of β-Arr2 full-length plasmid promotes PA-induced cardiomyocyte apoptosis, which remind us to focus on the changes of its location. β-Arr2 siRNA decreased the background level of β-Arr2 in nucleus in normal H9c2 cells. Overexpression of β-Arr2 increased cytoplasm level of β-Arr2 as PA did. While LiCl, the inhibitor of GSK-3β decreased PA-induced apoptosis, accompany with increased nucleus level of β-Arr2. Then we concluded that GSK-3β is closely associated with cardiomyocyte apoptosis induced by PA, it performs its pro-apoptotic function by affecting the location of β-Arr2. LiCl inhibits PA-induced cardiomyocyte apoptosis, which might provide novel therapeutic for cardiovascular diseases induced by .

Keyword: metabolic syndrome

Renal Lipotoxicity-Associated Inflammation and Insulin Resistance Affects Actin Cytoskeleton Organization in Podocytes.

In the last few decades a change in lifestyle has led to an alarming increase in the prevalence of obesity and obesity-associated complications. Obese patients are at increased risk of developing hypertension, heart disease, insulin resistance (IR), dyslipidemia, type 2 diabetes and renal disease. The excess calories are stored as triglycerides in adipose tissue, but also may accumulate ectopically in other organs, including the kidney, which contributes to the damage through a toxic process named lipotoxicity. Recently, the evidence suggests that renal lipid accumulation leads to glomerular damage and, more specifically, produces dysfunction in podocytes, key cells that compose and maintain the glomerular filtration barrier. Our aim was to analyze the early mechanisms underlying the development of renal disease associated with the process of lipotoxicity in podocytes. Our results show that treatment of podocytes with produced intracellular accumulation of lipid droplets and abnormal glucose and lipid metabolism. This was accompanied by the development of inflammation, oxidative stress and endoplasmic reticulum stress and insulin resistance. We found specific rearrangements of the actin cytoskeleton and slit diaphragm proteins (Nephrin, P-Cadherin, Vimentin) associated with this insulin resistance in -treated podocytes. We conclude that lipotoxicity accelerates glomerular disease through lipid accumulation and inflammation. Moreover, saturated fatty acids specifically promote insulin resistance by disturbing the cytoarchitecture of podocytes. These data suggest that renal lipid metabolism and cytoskeleton rearrangements may serve as a target for specific therapies aimed at slowing the progression of podocyte failure during .

Keyword: metabolic syndrome

pathways of oleic and are intensified in PCOS patients with normal androgen levels.

The aetiology and pathogenesis polycystic ovary (PCOS) remain uncertain and thus the relative studies are still crucial.Our aim was to analyse the fatty acids profiles of the main phospholipids species in serum from women with PCOS classified into phenotypes, and to diagnose women more susceptible to the occurrence of inflammatory state.PCOS screening tests were performed in The Clinic of Gynecology and Urogynecology of Pomeranian Medical University in the 2014-2015 years.The study are designed for general community and a primary care or referral center.39 patients with PCOS, diagnosed according to Rotterdam\'s criteria, and 14 healthy women, as a control group, participated in this study. Fatty profiles were investigated using gas chromatography. A total of 36 fatty acids and their derivatives were identified and quantified.Changes in fatty acids profile in plasma from women with PCOS phenotypes are not identical.The analyses showed lowered level of total SFA, increase in the concentration of caprylic and the activation of and oleic acids pathways. The level of nervonic was several times higher than in the control group, and the levels of behenic and tricosanoic acids were reduced.In both phenotypes the alternative pathways of oleic were activated, but they were more pronounced in women with proper level of androgens. Gamma-linolenic (C18:3n6) can be a factor protecting hyperandrogenic women.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: metabolic syndrome

Depot-specific differences in fatty composition and distinct associations with lipogenic gene expression in abdominal adipose tissue of obese women.

Cardiometabolic diseases are primarily linked to enlarged visceral adipose tissue (VAT). However, some data suggest heterogeneity within the subcutaneous adipose tissue (SAT) depot with potential differences between the superficial SAT (sSAT) and deep SAT (dSAT) compartments. We aimed to investigate the heterogeneity of these three depots with regard to fatty (FA) composition and gene expression. Adipose tissue biopsies were collected from 75 obese women undergoing laparoscopic gastric bypass surgery. FA composition and gene expression were determined with gas chromatography and quantitative real-time-PCR, respectively. Stearoyl CoA desaturase-1 (SCD-1) activity was estimated by product-to-precursor FA ratios. All polyunsaturated FAs (PUFA) with 20 carbons were consistently lower in VAT than either SAT depots, whereas essential PUFA (linoleic , 18:2n-6 and α-linolenic , 18:3n-3) were similar between all three depots. Lauric and were higher and lower in VAT, respectively. The SCD-1 product palmitoleic as well as estimated SCD-1 activity was higher in VAT than SAT. Overall, there was a distinct association pattern between lipid metabolizing genes and individual FAs in VAT. In conclusion, SAT and VAT are two distinct depots with regard to FA composition and expression of key lipogenic genes. However, the small differences between sSAT and dSAT suggest that FA metabolism of SAT is rather homogenous.

Keyword: metabolic syndrome

Substrate-energy metabolism and risk factors for cardiovascular disease in relation to fetal growth and adult body composition.

The effect of fetal programming on intermediary metabolism is uncertain. Therefore, we examined whether fetal programming affects oxidative and nonoxidative macronutrient metabolism and the prevalence of the in adult life. Healthy older men, aged 64-72 years, with either a lower birth weight (LBW, or=75th %ile; n = 13) had measurements of 1) net oxidative metabolism using indirect calorimetry before and for 6 h after a mixed meal (3,720 kJ) and 2) postprandial oxidation of exogenous [13C]. Body composition was measured using dual-energy X-ray absorptiometry. After adjustment for current weight and height, the LBW group had a lower resting energy expenditure (REE) in the preprandial (4.01 vs. 4.54 kJ/min, P = 0.015) and postprandial state (4.60 vs. 5.20 kJ/min, P = 0.004), and less fat-free mass than the HBW group. The BW category was a significant, independent, and better predictor of REE than weight plus height. There were no significant differences between groups in net oxidative and nonoxidative macronutrient (protein, fat, carbohydrate) metabolism (or of exogenous [13C]palmitate) or in the prevalence of the , which was present almost twice as commonly in the LBW than in the HBW group. The study suggests that fetal programming affects both pre- and postprandial EE in older life by mechanisms that are at least partly related to the mass of the fat-free body. BW was found to be a significant predictor of REE that was independent of adult weight plus height.

Keyword: metabolic syndrome

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free fatty (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: metabolic syndrome

induces osteoblastic differentiation in vascular smooth muscle cells through ACSL3 and NF-κB, novel targets of eicosapentaenoic .

Free fatty acids (FFAs), elevated in and diabetes, play a crucial role in the development of atherosclerotic cardiovascular disease, and eicosapentaenoic (EPA) counteracts many aspects of FFA-induced vascular pathology. Although vascular calcification is invariably associated with atherosclerosis, the mechanisms involved are not completely elucidated. In this study, we tested the hypothesis that EPA prevents the osteoblastic differentiation and mineralization of vascular smooth muscle cells (VSMC) induced by (PA), the most abundant long-chain saturated fatty in plasma. PA increased and EPA abolished the expression of the genes for bone-related proteins, including bone morphogenetic protein (BMP)-2, Msx2 and osteopontin in human aortic smooth muscle cells (HASMC). Among the long-chain acyl-CoA synthetase (ACSL) subfamily, ACSL3 expression was predominant in HASMC, and PA robustly increased and EPA efficiently inhibited ACSL3 expression. Importantly, PA-induced osteoblastic differentiation was mediated, at least in part, by ACSL3 activation because acyl-CoA synthetase (ACS) inhibitor or siRNA targeted to ACSL3 completely prevented the PA induction of both BMP-2 and Msx2. Conversely, adenovirus-mediated ACSL3 overexpression enhanced PA-induced BMP-2 and Msx2 expression. In addition, EPA, ACSL3 siRNA and ACS inhibitor attenuated calcium deposition and caspase activation induced by PA. Notably, PA induced activation of NF-κB, and NF-κB inhibitor prevented PA-induction of osteoblastic gene expression and calcium deposition. Immunohistochemistry revealed the prominent expression of ACSL3 in VSMC and macrophages in human non-calcifying and calcifying atherosclerotic plaques from the carotid arteries. These results identify ACSL3 and NF-κB as mediators of PA-induced osteoblastic differentiation and calcium deposition in VSMC and suggest that EPA prevents vascular calcification by inhibiting such a new molecular pathway elicited by PA.

Keyword: metabolic syndrome

Effect of Euterpe oleracea Mart. (Açaí) Oil on Dyslipidemia Caused by Cocos nucifera L. Saturated Fat in Wistar Rats.

Dyslipidemia is caused by disturbances in lipid metabolism that lead to chronic elevations of serum lipids, especially low-density lipoprotein (LDL)-cholesterol and triglycerides, increasing the risk of , obesity, diabetes, atherogenic processes, and cardiovascular diseases. The oil from the fruits of Euterpe oleracea (OFEO) is rich in unsaturated fatty acids with potential for treating alterations in lipid metabolism. In this study, we aimed to investigate the effect of OFEO on hyperlipidemia induced by Cocos nucifera L. saturated fat (GSC) in Wistar rats. Chromatographic profile showed that unsaturated fatty acids account for 66.08% in OFEO, predominately oleic (54.30%), and saturated fatty acids ( 31.6%) account for 33.92%. GSC-induced dyslipidemia resulted in an increase in total cholesterol, LDL-cholesterol, triglycerides, glucose, and liver and abdominal fat, as well as atherogenic processes in the thoracic aorta. OFEO treatment did not reduce hypertriglyceridemia, but did reduce total cholesterol and LDL-cholesterol, thus contributing to the antiatherogenic action of OFEO. OFEO treatment inhibited the formation of atheromatous plaques in the vascular endothelium of the treated rats, as well as those who were treated with simvastatin. The results obtained suggest that OFEO has an antiatherogenic effect in a rat model of dyslipidemia.

Keyword: metabolic syndrome

Synthesis and biological evaluations of marine oxohexadecenoic acids: PPARα/γ dual agonism and anti-diabetic target gene effects.

Obesity and associated disorders such as and type 2 diabetes (T2D) have reached epidemic proportions. Several natural products have been reported as Peroxisome Proliferator-Activated Receptor (PPAR) agonists, functioning as lead compounds towards developing new anti-diabetic drugs due to adverse side effects of existing PPAR drugs. We recently isolated and identified (7E)-9-oxohexadec-7-enoic (1) and (10E)-9-oxohexadec-10-enoic (2) from the marine algae Chaetoceros karianus. Herein we report the total synthesis, pharmacological characterization, and biological evaluations of these naturally occurring oxo-fatty acids (oFAs). The syntheses of 1 and 2 afforded sufficient material for extensive biological evaluations. Both oFAs show an appreciable dose-dependent activation of PPARα and -γ, with EC values in the micromolar range, and an ability to regulate important PPAR target genes in hepatocytes and adipocytes. Moreover, both 1 and 2 are able to drive adipogenesis when evaluated in the Simpson-Golabi-Behmel (SGBS) pre-adipocyte cell model, but with lowered expression of adipocyte markers and reduced lipid accumulation compared to the drug rosiglitazone. This seems to be caused by a transient upregulation of PPARγ and C/EBPα expression. Importantly, whole transcriptome analysis shows that both compounds induce anti-diabetic gene programs in adipocytes by upregulating insulin-sensitizing adipokines and repressing pro-inflammatory cytokines.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: metabolic syndrome

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the liver was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce imbalances typical of the and obesity in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

Keyword: metabolic syndrome

effects of fatty -bearing albumin on a proximal tubule cell line.

In glomerular disease, fatty acids carried on albumin are taken up by the proximal tubule with filtered albumin. We postulate that the fatty acids carried on filtered albumin could contribute to the deleterious effects of proteinuria. The effects of fatty -albumin complexes on lipid metabolism have been studied in opossum kidney (OK) cells, a proximal tubule cell line. OK cells transported two-thirds of [14C]palmitate-albumin (5 mg/ml) intracellularly within 16 h. [14C]palmitate-albumin was distributed into phosphatidylcholines, phosphatidylinositols, and tri- and diglycerides. 14C-labeled unsaturated fatty albumins (oleate, linoleate, and arachidonate) showed preferential incorporation into triglycerides, with lesser incorporation into phospholipids. Studies of total lipid pools showed that fatty -albumin uptake produced a particularly marked increase in total triglyceride levels (approximately 10-fold). Oil red O staining of OK cells cultured with oleate-albumin showed a marked increase in intracellular lipid droplets, compared with cells cultured with delipidated albumin, consistent with triglyceride accumulation. Less than 1% of [14C]palmitate taken up was isolated as intracellular free fatty . Less than 5% of [14C]palmitate internalized was oxidized to 14CO2. Different fatty acids, when taken up by the OK cell, have distinct fates. Each fatty is incorporated in a characteristic fashion into certain complex lipids, possibly dependent on the presence or absence of double bonds. We propose that this may have functional consequences for the proximal tubule in the human nephrotic .

Keyword: metabolic syndrome

Identification of Potential Biomarkers for Urine Metabolomics of Polycystic Ovary Based on Gas Chromatography-Mass Spectrometry.

Polycystic ovary (PCOS) is a complex endocrine and disorder, and it\'s diagnosis is difficult. The aim of this study was to investigate the profiles of PCOS patients by analyzing urine samples and identify useful biomarkers for diagnosis of PCOS.This study was carried out in the Department of Obstetrics and Gynecology of the Maternal and Child Health Hospital of Hunan Province from December 2014 to July 2016. In this study, the urine samples of 21 women with PCOS and 16 healthy controls were assessed through gas chromatography-mass spectrometry to investigate the urine metabolite characteristics of PCOS and identify useful biomarkers for the diagnosis of this disorder. The Student\'s t-test and rank sum test were applied to validate the statistical significance of the between the two groups.In total, 35 urine metabolites were found to be significantly different between the PCOS patients and the controls. In particular, a significant increase in the levels of lactose (10.01 [0,13.99] mmol/mol creatinine vs. 2.35 [0.16, 3.26] mmol/mol creatinine, P = 0.042), stearic (2.35 [1.47, 3.14] mmol/mol creatinine vs. 0.05 [0, 0.14] mmol/mol creatinine, P < 0.001), and (2.13 [1.07, 2.79] mmol/mol creatinine vs. 0 [0, 0] mmol/mol creatinine, P < 0.001) and a decrease in the levels of succinic (0 [0, 0] mmol/mol creatinine vs. 38.94 [4.16, 51.30] mmol/mol creatinine, P < 0.001) were found in the PCOS patients compared with the controls. It was possible to cluster the PCOS patients and the healthy controls into two distinct regions based on a principal component analysis model. Of the differentially expressed metabolites, four compounds, including stearic , , benzoylglycine, and threonine, were selected as potential biomarkers.This study offers new insight into the pathogenesis of PCOS, and the discriminating urine metabolites may provide a prospect for the diagnosis of PCOS.

Keyword: metabolic syndrome

Prolonged QT interval and lipid alterations beyond β-oxidation in very long-chain acyl-CoA dehydrogenase null mouse hearts.

Patients with very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency frequently present cardiomyopathy and heartbeat disorders. However, the underlying factors, which may be of cardiac or extra cardiac origins, remain to be elucidated. In this study, we tested for and functional alterations in the heart from 3- and 7-mo-old VLCAD null mice and their littermate counterparts, using validated experimental paradigms, namely, 1) ex vivo perfusion in working mode, with concomitant evaluation of myocardial contractility and fluxes using (13)C-labeled substrates under various conditions; as well as 2) in vivo targeted lipidomics, gene expression analysis as well as electrocardiogram monitoring by telemetry in mice fed various diets. Unexpectedly, when perfused ex vivo, working VLCAD null mouse hearts maintained values similar to those of the controls for functional parameters and for the contribution of exogenous palmitate to β-oxidation (energy production), even at high palmitate concentration (1 mM) and increased energy demand (with 1 μM epinephrine) or after fasting. However, in vivo, these hearts displayed a prolonged rate-corrected QT (QTc) interval under all conditions examined, as well as the following lipid alterations: 1) age- and condition-dependent accumulation of triglycerides, and 2) 20% lower docosahexaenoic (an omega-3 polyunsaturated fatty ) in membrane phospholipids. The latter was independent of liver but affected by feeding a diet enriched in saturated fat (exacerbated) or fish oil (attenuated). Our finding of a longer QTc interval in VLCAD null mice appears to be most relevant given that such condition increases the risk of sudden cardiac death.

Keyword: metabolic syndrome

Ceramide stearic to ratio predicts incident diabetes.

Ceramide lipids have a role in the development of insulin resistance, diabetes and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident diabetes.A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the (Prevent [PrevMetSyn], n\u2009=\u2009371). Diabetes risk score models were developed to estimate the 10\xa0year risk of incident diabetes.Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident diabetes (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and diabetes risk scores significantly decreased in individuals who had 5% or more weight loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident diabetes, and may be modulated by lifestyle intervention.

Keyword: metabolic syndrome

Histone methyltransferase G9a modulates hepatic insulin signaling via regulating HMGA1.

Hepatic insulin sensitivity is critical for glucose homeostasis, and insulin resistance is a fundamental found in various disorders, including obesity and type 2 diabetes. Despite considerable studies on the mechanisms of hepatic insulin resistance, the link between epigenetic regulation and the development of insulin resistance remains elusive. Here, we reported that G9a/EHMT2, a histone methyltransferase, was markedly decreased in the liver of db/db mice and high-fat diet (HFD)-fed mice. In cultured hepatic cells, G9a knockdown resulted in downregulation of insulin receptor, p-AKT and p-GSK3β; while upon upregulation, G9a prevented the - or glucosamine-induced insulin resistance by preserving the normal level of insulin receptor and integrity of insulin signaling. Further mechanistic study suggested that G9a regulated the expression level of high mobility group AT-hook 1 (HMGA1), a key regulator responsible for the transcription of insulin receptor (INSR) gene. Overexpression of HMGA1 normalized the impaired insulin signaling in G9a knockdown hepatic cells. Importantly, in db/db mice, restoring the expression level of G9a not only upregulated HMGA1 level and improved the impaired hepatic insulin signaling, but also alleviated hyperglycemia and hyperinsulinemia. Together, our results revealed a novel role for G9a in modulating insulin signaling, at least in part, depending on its regulatory function on HMGA1.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Soraphen, an inhibitor of the acetyl-CoA carboxylase system, improves peripheral insulin sensitivity in mice fed a high-fat diet.

Inhibition of the acetyl-CoA carboxylase (ACC) system, consisting of the isozymes ACC1 and ACC2, may be beneficial for treatment of insulin resistance and/or obesity by interfering with de novo lipogenesis and beta-oxidation. We have evaluated effects of pharmacological inhibition of ACC by soraphen (SP) on high fat (HF) diet-induced insulin resistance in mice.Male C57Bl6/J mice were fed control chow, a HF diet or a HF diet supplemented with SP (50 or 100 mg/kg/day).Body weight gain and total body fat content of SP-treated animals were significantly reduced compared with HF-fed mice. Fractional synthesis of palmitate was significantly reduced in mice treated with SP, indicative for ACC1 inhibition. Plasma beta-hydroxybutyrate levels were significantly elevated by SP, reflecting simultaneous inhibition of ACC2 activity. Mice treated with SP showed improved peripheral insulin sensitivity, as assessed by hyperinsulinaemic euglycaemic clamps.Pharmacological inhibition of the ACC system is of potential use for treatment of key components of the .

Keyword: metabolic syndrome

Free fatty acids inhibit insulin signaling-stimulated endothelial nitric oxide synthase activation through upregulating PTEN or inhibiting Akt kinase.

In , a systemic deregulation of the insulin pathway leads to a combined deregulation of insulin-regulated metabolism and cardiovascular functions. Free fatty acids (FFAs), which are increased in , inhibit insulin signaling and induce insulin resistance. This study was designed to examine FFAs\' effects on vascular insulin signaling and endothelial nitric oxide (NO) synthase (eNOS) activation in endothelial cells. We showed that FFAs inhibited insulin signaling and eNOS activation through different mechanisms. While linoleic inhibited Akt-mediated eNOS phosphorylation, appeared to affect the upstream signaling. Upregulation of PTEN (phosphatase and tensin homolog deleted on chromosome 10) activity and transcription by mediated the inhibitory effects on insulin signaling. We further found that activated stress signaling p38, but not Jun NH(2)-terminal kinase, was involved in PTEN upregulation. The p38 target transcriptional factor activating transcription factor (ATF)-2 bound to the PTEN promoter, which was increased by treatment. In summary, both and linoleic exert inhibitory effect on insulin signaling and eNOS activation in endothelial cells. inhibits insulin signaling by promoting PTEN activity and its transcription through p38 and its downstream transcription factor ATF-2. Our findings suggest that FFA-mediated inhibition of vascular insulin signaling and eNOS activation may contribute to cardiovascular diseases in .

Keyword: metabolic syndrome

Urinary metabonomic study of patients with acute coronary using UPLC-QTOF/MS.

This urinary metabonomic study aimed to identify the potential biomarkers in acute coronary (ACS) patients. Ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) was used to analyze the urine samples from ACS patients and healthy controls. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were applied to characterizing the endogenous metabolites and potential biomarker, respectively. Among twenty biomarkers that functioned in nine pathways, nine biomarkers were found up-regulated significantly, including of isobutyryl‑l‑carnitine, 3‑methylglutarylcarnitine, cinnavalininate, l‑tryptophan, 3‑methyldioxyindole, , N4‑acetylaminobutanal, 3‑sulfino‑l‑alanine and S‑adenosyl‑l‑homocysteine. The other eleven biomarkers were showed down-regulated, including of l‑lactic , trigonelline, nicotinuric , l‑alanine, d‑alanyl‑d‑alanine, creatine, N4‑acetylaminobutanoate, glutathionyl spermidine, 5‑methoxytryptamine, kynurenic and xanthurenic . This study also implied that fatty metabolism, fatty β‑oxidation metabolism, amino metabolism and TCA cycle played important roles in ACS. Therefore, urinary metabolomics may improve the diagnosis efficacy of ACS and make it more accurate and comprehensive for ACS diagnosis.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Stearoyl-CoA desaturase-1 (SCD1) augments saturated fatty -induced lipid accumulation and inhibits apoptosis in cardiac myocytes.

Mismatch between the uptake and utilization of long-chain fatty acids in the myocardium leads to abnormally high intracellular fatty concentration, which ultimately induces myocardial dysfunction. Stearoyl-Coenzyme A desaturase-1 (SCD1) is a rate-limiting enzyme that converts saturated fatty acids (SFAs) to monounsaturated fatty acids. Previous studies have shown that SCD1-deficinent mice are protected from insulin resistance and diet-induced obesity; however, the role of SCD1 in the heart remains to be determined. We examined the expression of SCD1 in obese rat hearts induced by a sucrose-rich diet for 3 months. We also examined the effect of SCD1 on myocardial energy metabolism and apoptotic cell death in neonatal rat cardiac myocytes in the presence of SFAs. Here we showed that the expression of SCD1 increases 3.6-fold without measurable change in the expression of lipogenic genes in the heart of rats fed a high-sucrose diet. Forced SCD1 expression augmented -induced lipid accumulation, but attenuated excess fatty oxidation and restored reduced glucose oxidation. Of importance, SCD1 substantially inhibited SFA-induced caspase 3 activation, ceramide synthesis, diacylglycerol synthesis, apoptotic cell death, and mitochondrial reactive oxygen species (ROS) generation. Experiments using SCD1 siRNA confirmed these observations. Furthermore, we showed that exposure of cardiac myocytes to glucose and insulin induced SCD1 expression. Our results indicate that SCD1 is highly regulated by a component in the heart, and such induction of SCD1 serves to alleviate SFA-induced adverse fatty catabolism, and eventually to prevent SFAs-induced apoptosis.

Keyword: metabolic syndrome

Identification of palmitate-regulated genes in HepG2 cells by applying microarray analysis.

Palmitate is the most abundant saturated fatty in the human diet and the major one synthesized de novo. To identify palmitate-regulated genes we performed whole genome mRNA expression profiling by using human hepatoma HepG2 cells. We identified eleven genes which are significantly (single-sided permutational t-test, p<0.05) regulated by low concentration of palmitate (50 microM). We observed a decreased expression of five metallothioneins, and an increased expression of liver expressed plasminogen activator inhibitor-1 protein and insulin-like growth factor II, which play a prominent role in the development of the . Comparative promoter analysis in-silico revealed common transcriptional regulation of differentially expressed genes through erythroid kruppel-like factor and members of the zinc binding protein factor family. In conclusion, low physiological palmitate concentrations changed expression of very responsive genes.

Keyword: metabolic syndrome

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: metabolic syndrome

Identification and synthesis of novel inhibitors of acetyl-CoA carboxylase with in vitro and in vivo efficacy on fat oxidation.

Acetyl CoA carboxylase isoforms 1 and 2 (ACC1/2) are key enzymes of fat utilization and their inhibition is considered to improve aspects of the . To identify pharmacological inhibitors of ACC1/2, a high throughput screen was performed which resulted in the identification of the lead compound 3 ( Gargazanli , G. ; Lardenois , P. ; Frost , J. ; George , P. Patent WO9855474 A1, 1998 ) as a moderate selective ACC2 inhibitor. Optimization of 3 led to 4m ( Zoller , G. ; Schmoll , D. ; Mueller , M. ; Haschke , G. ; Focken , I. Patent WO2010003624 A2, 2010 ) as a submicromolar dual ACC1/2 inhibitor of the rat and human isoforms. 4m possessed favorable pharmacokinetic parameters. This compound stimulated fat oxidation in vivo and reduced plasma triglyceride levels in a rodent model after subchronic administration. 4m is a suitable tool compound for the elucidation of the pharmacological potential of ACC1/2 inhibition.

Keyword: metabolic syndrome

Free fatty acids inhibit TM-EPCR expression through JNK pathway: an implication for the development of the prothrombotic state in .

is associated with significant hypercoagulable prothrombotic tendency; however, the mechanism for the prothrombotic state is not completely understood. We hypothesize that higher circulating plasma free fatty acids (FFAs) in inhibit the endothelial thrombomodulin (TM)-endothelial protein C receptor (EPCR) pathway, thereby promoting thrombus formation. Human umbilical vein endothelial cells were cultured in media supplemented with various doses of (PA), in the presence or absence of JNK inhibitor, and the expression of TM and EPCR was measured by western blot. The thrombotic state of high fat fed C57BL/6J mice was examined by tail bleeding time and deep venous thrombosis (DVT) model. As a result, PA inhibited the expression of TM and EPCR in endothelial cells, and this effect was blunted by inhibiting JNK signaling. High fat diet fed mice had higher level of circulating FFAs and exhibited prothrombotic state, evidenced by increased tail bleeding time and enlarged thrombotic size in DVT model, compared to the control diet fed mice. Hence, FFAs inhibit TM-EPCR-Protein C system in endothelial cells through activating JNK signaling, which may be a mechanism for the prothrombotic state in .

Keyword: metabolic syndrome

High glucose, unsaturated and saturated fatty acids differentially regulate expression of ATP-binding cassette transporters ABCA1 and ABCG1 in human macrophages.

The ATP-binding cassette transporters ABCA1 and ABCG1 are highly expressed in macrophage-derived foam cells and promote reverse cholesterol efflux via biogenesis of high-density lipoproteins. The aim of this study was to analyze the direct effects of bioactive factors related to the on macrophage transcript levels of all 47 human ABC transporters. Using in vitro M-CSF predifferentiated macrophages and TaqMan low density arrays we could show that linoleic , , and high glucose levels have a major impact on ABCA1 and ABCG1 expression but do not strongly affect most other human ABC transporters. In Western blot experiments we demonstrate that ABCA1 and ABCG1 protein levels are synchronously suppressed by high glucose levels and the w6-unsaturated fatty linoleic . We conclude that metabolites associated with the enhance the formation of atherosclerotic lesions by diminishing the reverse cholesterol transport function of ABCA1 and ABCG1.

Keyword: metabolic syndrome

is an intracellular signaling molecule involved in disease development.

Emerging evidence shows that (PA), a common fatty in the human diet, serves as a signaling molecule regulating the progression and development of many diseases at the molecular level. In this review, we focus on its regulatory roles in the development of five pathological conditions, namely, , cardiovascular diseases, cancer, neurodegenerative diseases, and inflammation. We summarize the clinical and epidemiological studies; and also the mechanistic studies which have identified the molecular targets for PA in these pathological conditions. Activation or inactivation of these molecular targets by PA controls disease development. Therefore, identifying the specific targets and signaling pathways that are regulated by PA can give us a better understanding of how these diseases develop for the design of effective targeted therapeutics.

Keyword: metabolic syndrome

Fatty Content of Plasma Triglycerides May Contribute to the Heterogeneity in the Relationship Between Abdominal Obesity and the .

About one-third of the people with abdominal obesity do not exhibit the (MetS). Fatty acids in plasma triglycerides (TGs) may help to explain part of this heterogeneity. This study compared TG fatty profile of adults with and without abdominal obesity and examined the associations of these fatty acids with MetS components.Fifty-four abdominally obese subjects were matched by age and sex with 54 adults without abdominal obesity. People were classified with MetS according to the harmonizing criteria for MetS. Fatty acids in plasma TGs were analyzed by gas chromatography.There were no differences in fatty acids of plasma TGs between people with and without abdominal obesity. However, there were differences between abdominally obese people with and without MetS. The abdominally obese group with MetS had higher (+2.9%; P\u2009=\u20090.012) and oleic (+4.0%; P\u2009=\u20090.001) acids and lower linoleic (-6.4%; P\u2009=\u20090.018) and arachidonic (-1.2%; P\u2009=\u20090.004) acids. After adjustment for abdominal obesity, age, and sex, a stepwise regression analysis showed that positively contributed to the variance in insulin (β\u2009=\u2009+1.08\u2009±\u20091.01; P\u2009=\u20090.000) and homeostasis model assessment of insulin resistance (HOMA-IR) index (β\u2009=\u2009+1.09\u2009±\u20091.01; P\u2009=\u20090.000) and myristic positively contributed to the variance in systolic blood pressure (β\u2009=\u2009+1.09\u2009±\u20091.03; P\u2009=\u20090.006). In contrast, linoleic negatively contributed to the variance in glucose (β\u2009=\u2009-0.321\u2009±\u20090.09; P\u2009=\u20090.001) and high-sensitivity C-reactive protein (hsCRP; β\u2009=\u2009-1.05\u2009±\u20091.01; P\u2009=\u20090.000).There were no differences in the plasma TG fatty profile between people with and without abdominal obesity. Likewise, fatty acids in plasma TGs associated with many of the MetS variables independently of abdominal obesity. These results suggest that the plasma TG fatty profile may help to explain part of the heterogeneity between abdominal obesity and the MetS.

Keyword: metabolic syndrome

Association of fatty composition in serum phospholipids with and arterial stiffness.

We examined the association of fatty (FA) composition in serum phospholipids with the features of (MetS) and arterial stiffness.Korean men (n = 593, 30-79 yrs) were categorized based on the number of MetS risk factors (RFs) and measured for the markers of MetS, serum phospholipid FA composition and brachial-ankle pulse wave velocity (baPWV), an index for the severity of arterial stiffness.Insulin resistance (HOMA-IR), baPWV, LDL size, and adiponectin were significantly altered corresponding to the number of MetS RFs. The proportions of total monounsaturated FA, palmitoleic (16:1), oleic (18:1ω-9) and dihomo-γ-linolenic (DGLA, 20:3ω-6) in serum phospholipids, and DGLA/linoleic (LA) (20:3ω-6/18:2ω-6), deta9-desaturase activity (D9D-16: 16:1/16:0 and D9D-18: 18:1ω-9/18:0) significantly increased corresponding to the number of MetS RFs, but D5D (20:4ω-6/20:3ω-6) decreased. baPWV positively correlated with HOMA-IR, (16:0), oleic , D6D (18:3ω-6/18:2ω-6), DGLA/LA and D9D-18, and negatively with adiponectin, LDL size, LA, docosahexaenoic (DHA, 22:6ω-3) and D5D. Multiple stepwise regression models revealed that baPWV was significantly influenced by systolic blood pressure, age, body weight, triglyceride and LA in serum phospholipids (R(2) = 0.378). Interestingly, baPWV (1419 ± 1 cm/s) and MetS (22%) were highest in individuals with lower proportion of LA (< 12.361%) and higher proportion of DGLA (≥ 1.412%) in serum phospholipid FAs.The features of MetS significantly related to serum phosopholipid FA composition. Particularly, arterial stiffness was associated with LA additively together with DLGA. It may suggest a potential benefit of sufficient amounts of LA in serum or in diet can reduce cardiovascular risk.Copyright © 2011 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Acetyl-CoA from inflammation-induced fatty acids oxidation promotes hepatic malate-aspartate shuttle activity and glycolysis.

Hepatic is associated with inflammation, as inflammation stimulates the reprogramming of nutrient metabolism and hepatic mitochondria-generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and fatty acids, in the condition of inflammation is still unclear. Here, we used an acute inflammation model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic glycolysis and fatty oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs liver showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain glycolysis. With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived fatty oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by fatty oxidation promotes MAS activity and glycolysis via nonenzymatic acetylation during the inflammatory stress response.

Keyword: metabolic syndrome

Fructose and glucose combined with free fatty acids induce disorders in HepG2 cell: A new model to study the impacts of high-fructose/sucrose and high-fat diets in vitro.

This work investigated the underlying mechanism of high-fructose/sucrose and high-fat diets, which rapidly induce in vivo, via a new cell model.Glucose and/or fructose were used to induce the human hepatoma cell (HepG2) in the presence of , oleic , or combined fatty acids (CFA) for 24 h. The alterations in lipid and uric production, glucose metabolism, oxidative status, and related genes and proteins were monitored. The cell model that featured disorders was established by treatment of 10 mM glucose and 15 mM fructose plus 1 mM CFA. Results showed that mainly induced insulin resistance, oxidative stress, and triglyceride (TG) secretion, whereas oleic mainly contributed to intracellular TG. Fructose was mainly responsible for uric and cholesterol production. In addition, fructose synergistically elevated the intra- and extracellular TG and extracellular malonaldehyde with glucose and CFA. Regulations of genes and proteins associated with carbohydrate metabolism and lipogenesis partially explained the action of fructose in inducing the disorders in cell.The combination of glucose, fructose, and CFA could successfully induce disorders in HepG2 cells, including dyslipidemia, insulin resistance, hyperuricemia, and oxidative stress.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: metabolic syndrome

Withdrawal from high-carbohydrate, high-saturated-fat diet changes saturated fat distribution and improves hepatic low-density-lipoprotein receptor expression to ameliorate in rats.

The "lipid hypothesis" determined that saturated fatty (SFA) raises low-density lipoprotein cholesterol, thereby increasing the risk for (MetS). The aim of this study was to investigate the effect of subchronic withdrawal from a high-carbohydrate, high-saturated fat (HCHF) diet during MetS with reference to changes in deleterious SFA (C12:0, lauric ; C14:0, myristic ; C16:0, ; C18:0, stearic ) distribution in liver, white adipose tissue (WAT), and feces.MetS induced by prolonged feeding of an HCHF diet in Wistar albino rat is used as a model of human MetS. The MetS-induced rats were withdrawn from the HCHF diet and changed to a basal diet for final 4\xa0wk of the total experimental duration of 16\xa0wk. SFA distribution in target tissues and hepatic low-density lipoprotein receptor (LDLr) expression were analyzed.Analyses of changes in SFA concentration of target tissues indicate that C16:0 and C18:0 reduced in WAT and liver after withdrawal of the HCHF diet. There was a significant (P\xa0<\xa00.001) decrease in fecal C12:0 with HCHF feeding, which significantly (P\xa0<\xa00.01) increased after withdrawal of this diet. Also, an improvement in expression of hepatic LDLr was observed after withdrawal of HCHF diet.The prolonged consumption of an HCHF diet leads to increased SFA accumulation in liver and WAT, decreased SFA excretion, and reduced hepatic LDLr expression during MetS, which is prominently reversed after subchronic withdrawal of the HCHF diet. This can contribute to better understanding of the fate of dietary SFA during MetS and may apply to the potential reversal of complications by the simple approach of nutritional modification.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Potential Role of Free Fatty Acids in the Pathogenesis of Periodontitis and Primary Sjögren\'s .

Clinical studies have shown that disorders such as type 2 diabetes and dyslipidemia are associated with increased risk of oral-related diseases, such as periodontitis and Sjögren\'s . Although changes in the immune system are critical in both of these disorders and oral-related diseases, the mechanism underlying the interaction between these diseases remains largely unknown. Obesity and type 2 diabetes are known to be associated with higher concentrations of free fatty acids in blood. Among free fatty acids, saturated fatty acids such as have been demonstrated to induce inflammatory responses mainly via the innate immune systems, and to be involved in the pathogenesis of type 2 diabetes in tissues such as adipose tissue, liver, pancreas, and skeletal muscle. Here, we highlight recent advances in evidence for the potential involvement of in the pathogenesis of periodontitis and Sjögren\'s , and discuss the possibility that improvement of the lipid profile could be a new strategy for the treatment of these diseases.

Keyword: metabolic syndrome

Chronic intake of moderate fat-enriched diet induces fatty liver and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and , there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of disorders, such as overweight/obesity and . However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: metabolic syndrome

(16:0) competes with omega-6 linoleic and omega-3 ɑ-linolenic acids for FADS2 mediated Δ6-desaturation.

Sapienic , 16:1n-10 is the most abundant unsaturated fatty on human skin where its synthesis is mediated by FADS2 in the sebaceous glands. The FADS2 product introduces a double bond at the Δ6, Δ4 and Δ8 positions by acting on at least ten substrates, including 16:0, 18:2n-6, and 18:3n-3. Our aim was to characterize the competition for accessing FADS2 mediated Δ6 desaturation between 16:0 and the most abundant polyunsaturated fatty acids (PUFA) in the human diet, 18:2n-6 and 18:3n-3, to evaluate whether competition may be relevant in other tissues and thus linked to abnormalities associated with FADS2 or fatty levels. MCF7 cells stably transformed with FADS2 biosynthesize 16:1n-10 from exogenous 16:0 in preference to 16:1n-7, the immediate product of SCD highly expressed in cancer cell lines, and 16:1n-9 via partial β-oxidation of 18:1n-9. Increasing availability of 18:2n-6 or 18:3n-3 resulted in decreased bioconversion of 16:0 to 16:1n-10, simultaneously increasing the levels of highly unsaturated products. FADS2 cells accumulate the desaturation-elongation products 20:3n-6 and 20:4n-3 in preference to the immediate desaturation products 18:3n-6 and 18:4n-3 implying prompt/coupled elongation of the nascent desaturation products. MCF7 cells incorporate newly synthesized 16:1n-10 into phospholipids. These data suggest that excess 16:0 due to, for instance, de novo lipogenesis from high carbohydrate or alcohol consumption, inhibits synthesis of highly unsaturated fatty acids, and may in part explain why supplemental preformed EPA and DHA in some studies improves insulin resistance and other factors related to diabetes and aggravated by excess calorie consumption.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: metabolic syndrome

Saturated long-chain fatty acids activate inflammatory signaling in astrocytes.

This study describes the effects of long-chain fatty acids on inflammatory signaling in cultured astrocytes. Data show that the saturated fatty , as well as lauric and stearic , trigger the release of TNFα and IL-6 from astrocytes. Unsaturated fatty acids were unable to induce cytokine release from cultured astrocytes. Furthermore, the effects of on cytokine release require Toll-like receptor 4 rather than CD36 or Toll-like receptor 2, and do not depend on metabolism to palmitoyl-CoA. Inhibitor studies revealed that pharmacologic inhibition of p38 or p42/44 MAPK pathways prevents the pro-inflammatory effects of , whereas JNK and PI3K inhibition does not affect cytokine release. Depletion of microglia from primary astrocyte cultures using the lysosomotropic agent l-leucine methyl ester revealed that the ability of to trigger cytokine release is not dependent on the presence of microglia. Finally, data show that the essential ω-3 fatty docosahexaenoic acts in a dose-dependent manner to prevent the actions of on inflammatory signaling in astrocytes. Collectively, these data demonstrate the ability of saturated fatty acids to induce astrocyte inflammation in vitro. These data thus raise the possibility that high levels of circulating saturated fatty acids could cause reactive gliosis and brain inflammation in vivo, and could potentially participate in the reported adverse neurologic consequences of obesity and .© 2012 The Authors. Journal of Neurochemistry © 2012 International Society for Neurochemistry.

Keyword: metabolic syndrome

Short-term Regulation of Resistin in vivo by Oral Lipid Ingestion and in vitro by Fatty Stimulation.

Dysbalance of pro- and anti-inflammatory adipokines is a hallmark of but their nutrition-dependent regulation in healthy individuals is poorly characterized. We investigated pro-inflammatory resistin and anti-inflammatory adiponectin regulation during oral lipid ingestion (OLI) in healthy adults. Response of resistin upon free fatty (FFA) stimulation was investigated in 3T3-L1 adipocytes.100 healthy volunteers underwent OLI. Venous blood was drawn after 0, 2, 4, and 6\u2009hours (h). Subjects were characterized by anthropometric and standard laboratory parameters. Serum concentrations of adiponectin and resistin were measured by enzyme-linked immunosorbent assay (ELISA). Adipocytes were stimulated with FFA and concentrations of adipokines were measured by ELISA.Irrespective of BMI and gender, OLI led to a significant reduction of resistin serum levels in a stepwise manner whereas adiponectin concentrations remained unchanged. There were positive correlations of resistin with waist/hip ratio and visfatin levels, as was calculated by regression analysis. Resistin concentrations were significantly higher in smokers when compared to non-smokers. Adiponectin concentrations were higher in females and in users of hormonal contraception. Adiponectin levels showed a positive correlation with heart rate and HDL cholesterol and a negative correlation with age, waist/hip-ratio, BMI, diastolic/systolic blood pressure, visfatin levels and LDL/HDL-ratio. Resistin secretion was significantly induced by , linoleic and oleic in adipocytes.OLI is a physiological repressor of systemic resistin release whereas FFA upregulate resistin release in vitro from adipocytes.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: metabolic syndrome

Ceramide-activated phosphatase mediates fatty -induced endothelial VEGF resistance and impaired angiogenesis.

Endothelial dysfunction, including endothelial hyporesponsiveness to prototypical angiogenic growth factors and eNOS agonists, underlies vascular pathology in many dysmetabolic states. We investigated effects of a saturated free fatty , (PA), on endothelial cell responses to VEGF. PA-pretreated endothelial cells had markedly diminished Akt, eNOS, and ERK activation responses to VEGF, despite normal VEGFR2 phosphorylation. PA inhibited VEGF-induced angiogenic cord formation in Matrigel, and PA-treated endothelial cells accumulated early species (C16) ceramide. The serine palmitoyltransferase inhibitor myriocin reversed these defects. Protein phosphatase 2A (PP2A) became more eNOS-associated in PA-treated cells; the PP2A inhibitor okadaic reversed PA-induced signaling defects. Mice fed a diet high in saturated fat for 2 to 3 weeks had impaired i) aortic Akt and eNOS phosphorylation to infused VEGF, ii) ear angiogenic responses to intradermal adenoviral-VEGF injection, and iii) vascular flow recovery to hindlimb ischemia as indicated by laser Doppler and αVβ3 SPECT imaging. High-fat feeding did not impair VEGF-induced signaling or angiogenic responses in mice with reduced serine palmitoyltransferase expression. Thus, de novo ceramide synthesis is required for these detrimental PA effects. The findings demonstrate an endothelial VEGF resistance mechanism conferred by PA, which comprises ceramide-induced, PP2A-mediated dephosphorylation of critical activation sites on enzymes central to vascular homeostasis and angiogenesis. This study defines potential molecular targets for preservation of endothelial function in .Copyright © 2014 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of obesity, insulin resistance, type-2 diabetes and . Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA metabolism on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: metabolic syndrome

Activation of PPARdelta up-regulates fatty oxidation and energy uncoupling genes of mitochondria and reduces palmitate-induced apoptosis in pancreatic beta-cells.

Recent evidence indicates that decreased oxidative capacity, lipotoxicity, and mitochondrial aberrations contribute to the development of insulin resistance and type 2 diabetes. The goal of this study was to investigate the effects of peroxisome proliferator-activated receptor delta (PPARdelta) activation on lipid oxidation, mitochondrial function, and insulin secretion in pancreatic beta-cells. After HIT-T15 cells (a beta-cell line) were exposed to high concentrations of palmitate and GW501516 (GW; a selective agonist of PPARdelta), we found that administration of GW increased the expression of PPARdelta mRNA. GW-induced activation of PPARdelta up-regulated carnitine palmitoyltransferase 1 (CPT1), long-chain acyl-CoA dehydrogenase (LCAD), pyruvate dehydrogenase kinase 4 (PDK4), and uncoupling protein 2 (UCP2); alleviated mitochondrial swelling; attenuated apoptosis; and reduced basal insulin secretion induced by increased palmitate in HIT cells. These results suggest that activation of PPARdelta plays an important role in protecting pancreatic beta-cells against aberrations caused by lipotoxicity in and diabetes.Copyright 2009 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Signature of MELAS/Leigh Overlap in Patient-specific Induced Pluripotent Stem Cells Model.

Mitochondrial myopathy, Encephalopathy, Lactic Acidosis, Stroke-like episodes/Leigh overlap (MELAS) is caused by defects in the mitochondrial respiratory chain. It is still largely unknown how these mitochondrial respiratory chain defects affect cellular metabolisms and lead to variable clinical phenotypes. Here, we analyzed signatures in a cellular model of MELAS/ Leigh overlap using untargeted gas chromatography coupled to mass spectrometry (GC-MS). .We obtained fibroblasts from a MELAS/Leigh overlap patient carrying the heteroplasmic m.10191T>C mutation, and generated induced pluripotent stem cells (iPSCs) from these fibroblast. Isogenic iPSC clones carrying two different loads of the heteroplasmic mutation (ND3hig-iPSC, ND3"*w- iPSC-) were subjected to metabolome analysis. Metabolite profiles, which were identified by GC-MS, were analyzed by principal component analysis (PCA).We were able to identify about 40 metabolites in control fibroblasts and iPSCs. Upon comparative metabolome analysis between fibroblasts and iPSCs, lactic and proline were distinct between the two groups. When we compared patient fibroblasts and control fibroblasts, no significant distinct metabolites were found. On the other hand, patient specific iPSC with high mutational load (ND3high_ iPSC) showed a distinct metabolite profile compared with ND3""-iPSC and control-iPSCs. Metabolites that contributed to this distinction were pyruvate, malic , , stearic , and lactic . This metabolomic signature was only seen in the undifferentiated state of iPSCs and was lost upon differentiationThese findings suggest that patient specific iPSC technology is useful to elucidate unique pathogenic pathways ,6mitochondrial chain diseases.

Keyword: metabolic syndrome

[Low and very low density lipoproteins: pathogenetic and clinical significance].

LDLP and VLDLP have different biological functions: phylogenetically older LDLP transfer FA that serve as substrates for intracellular production of energy and ATP while VLDLP transfer FA--precursors of cell membranes and eicosanoids. The cells absorb LDLP via apoB-100 endocytosis and VLDLP through apoE/B-100 receptors. VLDLP consist of and oleic VLDLP and LDLP of linoleic and linolenic LDLP. The contribution of LDLP to the development of HLP atherosclerosis and atheromatosis is negligible. LDLP form and oleic VLDLP with hydrated LDLP density. Blockade of LDLP absorption by apoB endocytosis and deficit of poly-FA constitute the etiological basis of atherosclerosis. Its pathogenetic basis is the excess of VLDLP with LDPL density in the intercellular space that block absorption of linoleic LDLP with all transferred SC poly-FA. Atheromatosis is clinically and prognostically most significant symptom of atherosclerosis associated with accumulation of ligand-free VLDLP and LDLP in arterial intima of the elastic type as the local pool of interstitial tissue for intravascular pool of intercellular medium. Type 2 diabetes mellitus in aged patients is a symptom of atherosclerosis resulting from SC poly-FA deficit and GLUT4 incompetence. Insulin-dependent cells differ in the degree of insulin resistance. Non-alcoholic fatty liver disease, synthesis of a physiological TG by hepatocytes and excessive formation of VLDLP in liver integrate pathogenesis of atherosclerosis and hepatic steatosis. The main pathogenetic factor is the excess of s-FA and TG.

Keyword: metabolic syndrome

Diet-induced obesity in the selenocysteine lyase knockout mouse.

Selenocysteine lyase (Scly) mediates selenocysteine decomposition. It was previously demonstrated that, upon adequate caloric intake (12% kcal fat) and selenium deficiency, disruption of Scly in mice leads to development of . In this study, we investigate the effect of a high-fat (45% kcal) selenium-adequate diet in Scly knockout (KO) mice on development of . Involvement of selenoproteins in energy metabolism after Scly disruption was also examined in vitro in the murine hepatoma cell line, Hepa1-6, following palmitate treatment.Scly KO mice were more susceptible to diet-induced obesity than their wild-type counterparts after feeding a high-fat selenium-adequate diet. Scly KO mice had aggravated hyperinsulinemia, hypercholesterolemia, glucose, and insulin intolerance, but unchanged inflammatory cytokines and expression of most selenoproteins, except increased serum selenoprotein P (Sepp1). Scly KO mice also exhibited enhanced hepatic levels of pyruvate and enzymes involved in the regulation of pyruvate cycling, such as pyruvate carboxylase (Pcx) and pyruvate dehydrogenase (Pdh). However, in vitro silencing of Scly in Hepa1-6 cells led to diminished Sepp1 expression, and concomitant palmitate treatment decreased Pdh expression.The role of selenium in lipid metabolism is recognized, but specific selenium-dependent mechanisms leading to obesity are unclear. This study uncovers that Scly has a remarkable effect on obesity and development triggered by high-fat exposure, independent of the expression of most selenoproteins.Diet-induced obesity in Scly KO mice is aggravated, with effects on pyruvate levels and consequent activation of energy metabolism independent of selenoprotein levels.

Keyword: metabolic syndrome

Calmodulin dependent protein kinase II activation by exercise regulates saturated & unsaturated fatty acids and improves some markers.

Activation of Calmodulin dependent protein kinase (CaMK)-II by exercise has a plethora of benefits in health. Fatty acids play a pivotal role in the pathogenesis of (MetS). Prevention of MetS and treatment of its main characteristics are very significant to fight against type 2 diabetes. CaMKII activation in the regulation of saturated and unsaturated fatty acids in relation to type 2 diabetes and MetS has not been studied, which became the focus of this present study.Using Gas chromatography-Mass spectrometry, we investigated saturated fatty acids and unsaturated fatty acids. Quantitative real time PCR was also used to assess the gene expression.Results indicate that both palmitoleic and oleic which are monounsaturated fatty acids were increased in response to CaMKII activation. On the other hand, myristic and which are saturated fatty acids known to increase the risk factors of MetS and type 2 diabetes were decreased by exercise induction of CaMKII. Conversely, lauric also a saturated fatty was increased in response to CaMKII activation by exercise. This fatty is known to have beneficial effects in alleviating symptoms of both type 2 diabetes and MetS.According to our knowledge, this is the first study to show that CaMKII activation by exercise regulates fatty acids essential in type 2 diabetes and MetS. CaMKII can be an avenue of designing novel therapeutic drugs in the management and treatment of type 2 diabetes and MetS.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Exacerbation and Prolongation of Psoriasiform Inflammation in Diabetic Obese Mice: A Synergistic Role of CXCL5 and Endoplasmic Reticulum Stress.

Accumulating evidence suggests that psoriasis is frequently accompanied by disorders, such as obesity and diabetes. However, the mechanisms underlying the association between increased psoriasis severity and concomitant have not been fully clarified. Herein, we show that imiquimod-induced psoriasiform inflammation was exacerbated and prolonged in diabetic obese mice compared to that in control mice, accompanied by remarkably increased lesional expressions of Cxcl5 and Il-1b. Notably, a large number of CXCL5 Ly6G cells infiltrated the dermis and subcutaneous fat tissue of the diabetic obese mice. Most macrophages in the subcutaneous fat tissues of the diabetic obese mice were positive for expression of IL-1β and GRP78/Bip, an endoplasmic reticulum stress marker. Depletion of Ly6G cells and macrophages diminished the imiquimod-induced psoriasiform inflammation. Further, CXCL5 potentiated the secretion of IL-1β from macrophages and , a fatty released from subcutaneous adipocytes, further enhanced IL-1β secretion via endoplasmic reticulum stress induction. Combined with the fact that the serum levels of both CXCL5 and are significantly elevated in patients with , our results suggest a role for CXCL5 and endoplasmic reticulum stress in the increase of psoriasis severity of patients with concomitant .Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Uncoupling AMPK from autophagy: a foe that hinders the beneficial effects of metformin treatment on -associated atherosclerosis? Focus on "glucose and palmitate uncouple AMPK from autophagy in human aortic endothelial cells".

Keyword: metabolic syndrome

Accumulation of lipids and oxidatively damaged DNA in hepatocytes exposed to particles.

Exposure to particles has been suggested to generate hepatosteatosis by oxidative stress mechanisms. We investigated lipid accumulation in cultured human hepatocytes (HepG2) and rat liver after exposure to four different carbon-based particles. HepG2 cells were exposed to particles for 3h and subsequently incubated for another 18h to manifest lipid accumulation. In an animal model of we investigated the association between intake of carbon black (CB, 14nm) particles and hepatic lipid accumulation, inflammation and gene expression of Srebp-1, Fasn and Scd-1 involved in lipid synthesis. There was a concentration-dependent increase in intracellular lipid content after exposure to CB in HepG2 cells, which was only observed after co-exposure to oleic/. Similar results were observed in HepG2 cells after exposure to diesel exhaust particles, fullerenes C60 or pristine single-walled carbon nanotubes. All four types of particles also generated oxidatively damaged DNA, assessed as formamidopyrimidine DNA glycosylase (FPG) sensitive sites, in HepG2 cells after 3h exposure. The animal model of showed increased lipid load in the liver after one oral exposure to 6.4mg/kg of CB in lean Zucker rats. This was not associated with increased iNOS staining in the liver, indicating that the oral CB exposure was associated with hepatic steatosis rather than steatohepatitis. The lipid accumulation did not seem to be related to increased lipogenesis because there were unaltered gene expression levels in both the HepG2 cells and rat livers. Collectively, exposure to particles is associated with oxidative stress and steatosis in hepatocytes.© 2013. Published by Elsevier Inc. All rights reserved.

Keyword: metabolic syndrome

Unsaturated fatty acids and insulin resistance in childhood obesity.

Obesity is characterized by increased levels of plasma free fatty acids (FFAs) that interfere with insulin signaling. The aim of our study was to assess the FFA profile in obese children and adolescents and to determine their relation with different degrees of insulin resistance.A transversal study was conducted of 51 children and adolescents (mean age, 11.7±1.6 years; 47% males) with obesity (body mass index ≥95 percentile). Anthropometric, clinical, and biochemical parameters were assessed. Insulin resistance was determined using the homeostasis model assessment of insulin resistance (HOMA-IR) index. Plasma fatty acids were quantified by high-performance liquid chromatography with heptadecanoic as the internal standard.The mean concentration of myristic , linoleic , , oleic , stearic , and total fatty acids was 9.3±2.2, 86.5±38.3, 93.0±35.5, 177.0±83.6, 48.5±14.9, and 414.3±160.9 μmol/L, respectively. Total fatty acids and unsaturated fatty acids such as oleic and linoleic showed an inverse significant correlation with insulin resistance. Children with high insulin resistance (HOMA-IR >2.5) showed a decrease in unsaturated fatty acids compared with children having a HOMA-IR of <2.5. There were no changes in saturated fatty concentrations between those groups.A decrease in unsaturated fatty acids was correlated with insulin resistance in childhood obesity.

Keyword: metabolic syndrome

Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis.

Autophagy, which is critical for the proper turnover of organelles such as endoplasmic reticulum and mitochondria, affects diverse aspects of metabolism, and its dysregulation has been incriminated in various disorders. However, the role of autophagy of myeloid cells in adipose tissue inflammation and type 2 diabetes has not been addressed. We produced mice with myeloid cell-specific deletion of Atg7 (autophagy-related 7), an essential autophagy gene (Atg7 conditional knockout [cKO] mice). While Atg7 cKO mice were metabolically indistinguishable from control mice, they developed diabetes when bred to ob/w mice (Atg7 cKO-ob/ob mice), accompanied by increases in the crown-like structure, inflammatory cytokine expression and inflammasome activation in adipose tissue. Mφs (macrophages) from Atg7 cKO mice showed significantly higher interleukin 1 β release and inflammasome activation in response to a plus lipopolysaccharide combination. Moreover, a decrease in the NAD(+):NADH ratio and increase in intracellular ROS content after treatment with in combination with lipopolysaccharide were more pronounced in Mφs from Atg7 cKO mice, suggesting that mitochondrial dysfunction in autophagy-deficient Mφs leads to an increase in lipid-induced inflammasome and deterioration in Atg7 cKO-ob/ob mice. Atg7 cKO mice were more susceptible to experimental colitis, accompanied by increased colonic cytokine expression, T helper 1 skewing and systemic bacterial invasion. These results suggest that autophagy of Mφs is important for the control of inflammasome activation in response to or extrinsic stress, and autophagy deficiency in Mφs may contribute to the progression of associated with lipid injury and colitis.

Keyword: metabolic syndrome

risk factors for stroke and transient ischemic attacks in middle-aged men: a community-based study with long-term follow-up.

The impact of lipometabolic and glucometabolic disturbances on stroke incidence remains to be characterized in detail. We investigated relations of a comprehensive panel of baseline lipometabolic and glucometabolic variables to incident fatal and nonfatal stroke or transient ischemic attack (TIA), and stroke subtypes.A community-based prospective study of 2313 middle-aged men invited to a health survey at age 50.During a follow-up of up to 32 years, 421 developed stroke or TIA. In Cox proportional hazards analyses adjusting for treatment with cardiovascular drugs at baseline, 1-standard deviation increases in body mass index, systolic and diastolic blood pressures, serum proinsulin, and lipoprotein(a) were associated with 11 to 35% increased risk for subsequent stroke/TIA. Electrocardiographic left ventricular hypertrophy and smoking were also associated with a higher risk for stroke/TIA. Essentially the same variables were related to brain infarction/TIA. Higher proportions of (16:0), palmitoleic (16:1), and oleic (18:1) in cholesterol esters were associated with an increased risk, whereas a higher proportion of linoleic (18:2 n-6) was protective against stroke/TIA. Further adjusting all models also for hypertension, diabetes, the , serum cholesterol, atrial fibrillation, cardiovascular disease, smoking, and physical activity, essentially the same pattern was observed.Indices of an unhealthy dietary fat intake and a high serum lipoprotein (a) level predicted fatal and nonfatal stroke/TIA independently of established risk factors in a community-based sample of middle-aged men followed for 32 years.

Keyword: metabolic syndrome

Skeletal muscle miR-34a/SIRT1:AMPK axis is activated in experimental and human non-alcoholic steatohepatitis.

Non-alcoholic fatty liver disease (NAFLD) pathogenesis associates with intramyocellular lipid deposition and mitochondrial dysfunction. microRNAs (miRs), including pro-apoptotic miR-34a, are modulated during disease progression in liver tissue and plasma. We aimed to investigate the functional role of the miR-34a/SIRT1:AMP-activated protein kinase (AMPK) pathway in modulating local mitochondrial dysfunction in the skeletal muscle of human and experimental non-alcoholic steatohepatitis. Muscle biopsies were obtained from morbid obese NAFLD patients undergoing bariatric surgery. C57BL/6N mice were fed different NAFLD-inducing diets and C2C12 muscle cells incubated with (PA) in the presence or absence of an AMPK activator, or upon miR-34a functional modulation. Several muscle miRNAs, including miR-34a, were found increased with human NAFLD progression. Activation of the miR-34a/SIRT1:AMPK pathway, concomitant with impairment in insulin signalling mediators and deregulation of mitochondrial-shaping proteins, was evident in C2C12 cells incubated with PA, as well as in the skeletal muscle of all three diet-induced NAFLD mice models. Functional studies established the association between miR-34a- and PA-induced muscle cell deregulation. Of note, activation of AMPK almost completely prevented miR-34a- and PA-induced cellular stress. In addition, the miR-34a/SIRT1:AMPK pathway and mitochondrial dynamics dysfunction were also found amplified in muscle of human NAFLD. Finally, muscle miR-34a expression and mitofusin 2 (Mfn2) protein levels correlated with hallmarks of NAFLD and disease progression. Our results indicate that activation of the miR-34a/SIRT1:AMPK pathway leads to mitochondrial dynamics dysfunction in skeletal muscle of human and experimental NAFLD, representing an appealing prospective target in . KEY MESSAGES: Skeletal muscle microRNAs are modulated during NAFLD progression. -induced muscle cell dysfunction occurs, at least in part, through activation of the miR-34a/SIRT1:AMPK pathway. miR-34a/SIRT1:AMPK activation associates with mitochondria dynamics dysfunction in human NAFLD.

Keyword: metabolic syndrome

Pigment epithelium-derived factor (PEDF) suppresses IL-1β-mediated c-Jun N-terminal kinase (JNK) activation to improve hepatocyte insulin signaling.

Pigment epithelium-derived factor (PEDF) is an antiinflammatory protein that circulates at high levels in the . studies of PEDF knockout (KO) mice were conducted to investigate the relationship between PEDF, inflammatory markers, and homeostasis. Male PEDF KO mice demonstrated a phenotype consisting of increased adiposity, glucose intolerance, and elevated serum levels of metabolites associated with the . Genome expression analysis revealed an increase in IL-1β signaling in the livers of PEDF KO mice that was accompanied by impaired IRS and Akt signaling. In human hepatocytes, PEDF blocked the effects of an IL-1β challenge by suppressing activation of the inflammatory mediator c-Jun N-terminal kinase while restoring Akt signaling. RNA interference of PEDF in human hepatocytes was permissive for c-Jun N-terminal kinase activation and decreased Akt signaling. A metabolomics profile identified elevated circulating levels of tricarboxyclic cycle intermediates including succinate, an inducer of IL-1β, in PEDF KO mice. Succinate-dependent IL-1β expression was blocked by PEDF in PEDF KO, but not wild-type hepatocytes. In vivo, PEDF restoration reduced hyperglycemia and improved hepatic insulin signaling in PEDF KO mice. These findings identify elevated PEDF as a homeostatic mechanism in the human .

Keyword: metabolic syndrome

α-Ionone attenuates high-fat diet-induced skeletal muscle wasting in mice via activation of cAMP signaling.

Skeletal muscle atrophy is a common and debilitating condition that lacks a pharmacological therapy. Our aim was to investigate the potential of α-ionone, a naturally occurring flavoring agent, in preventing muscle atrophy and to delineate the mechanisms involved. We found that α-ionone not only stimulated myogenesis but also attenuated -induced atrophy of cultured skeletal myotubes, as evidenced by an increased myotube diameter and length, fusion index, and cellular protein content. These beneficial actions of α-ionone were abrogated by cAMP inhibitor. The antiatrophic effects of α-ionone on cultured myotubes were confirmed in a corresponding mouse model. The skeletal muscle mass, muscle protein content, myofiber diameter, and muscle strength were greater in α-ionone-treated mice than in untreated animals fed high-fat diet. Furthermore, α-ionone increased cAMP concentration and enhanced its downstream PKA-CREB signaling in skeletal muscle of mice fed high-fat diet. Thus, α-ionone is a promising agent that may enhance skeletal muscle mass and strength.

Keyword: metabolism

Urinary metabolomics for discovering biomarkers of laryngeal cancer using UPLC-QTOF/MS.

The incidence of laryngeal cancer (LYC) is second only to lung cancer, which is also the second most common cancer in head and neck cancer. Risk assessment metabolomics biomarkers to diagnose LYC have not been found by now. To profile the metabolites in healthy controls (HCs) and LYC patients (LYCs), urine metabonomics study was performed based on reversed phase liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RPLC-QTOF/MS). Six urine differentially expressed metabolites (Variable Importance in Projection >1, p\u2009<\u2009 0.05) were identified by searching reference library or comparing with standard based on OPLS-DA (orthogonal partial least squares-discriminant analysis) model. d-pantothenic , , myristic , oleamide, sphinganine and phytosphingosine were identified as differential metabolites associated with the LYC and they might play roles in sphingolipid , fatty biosynthesis, fatty elongation in mitochondria, pantothenate and coenzyme A (CoA) biosynthesis, beta-Alanine and fatty . These six differential metabolites were combined to test the potentiality of diagnosis of LYC. Results revealed that the area under the curve (AUC) value, sensitivity and specificity of receiving operator characteristic (ROC) curve were 0.97, 95% and 97%, respectively, indicating that this diagnosis method could be used to distinguish LYCs from HCs with good sensitivity and specificity.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolism

Dietary zerumbone from shampoo ginger: new insights into its antioxidant and anticancer activity.

The dietary sesquiterpene dienone zerumbone (ZER) selectively targets cancer cells, inducing mitochondrial dysfunction and apoptosis, and protects non-cancerous cells towards oxidative stress and insult. This study examines the in vitro effects of ZER on lipid peroxidation in biological systems (cholesterol and phospholipid membrane oxidation) and explores its antitumor action in terms of its ability to modulate cancer cell lipid profile. Evaluation of the antioxidant activity of ZER showed that this compound is unable to trap lipoperoxyl radicals per se. ZER significantly modulated the total lipid and fatty profiles in cancer cells, inducing marked changes in the phospholipid/cholesterol ratio, with significant decreases in the levels of oleic and acids and a marked increase of stearic . Cell-based fluorescent measurements of intracellular membranes and lipid droplets using the Nile Red staining technique showed that in cancer cells, ZER induced significant accumulation of cytosolic lipid droplets and altered cell membrane organization/protein dynamics, depolarizing the mitochondrial membranes and inducing apoptosis and alteration of nuclear morphology. The modulatory activity of ZER on the total lipid and fatty profiles and lipid droplets may therefore represent another possible mechanism of its anticancer properties.

Keyword: metabolism

Overexpression of Acyl-ACP Thioesterases, and , Induce Distinct Gene Expression Reprogramming in Developing Seeds of .

We examined the substrate preference of acyl-ACP thioesterases, and , and gene expression changes associated with the modification of lipid composition in the seed, using transgenic plants overexpressing or under the control of a seed-specific promoter. seeds contained a higher level of total saturated fatty (FA) content, with 4.3 times increase in 16:0 , whereas seeds showed approximately 3% accumulation of 10:0 and 12:0 medium-chain FAs, and a small increase in other saturated FAs, resulting in higher levels of total saturated FAs. RNA-Seq analysis using entire developing pods at 8, 25, and 45 days after flowering (DAF) showed up-regulation of genes for β-ketoacyl-acyl carrier protein synthase I/II, stearoyl-ACP desaturase, oleate desaturase, and linoleate desaturase, which could increase unsaturated FAs and possibly compensate for the increase in 16:0 at 45 DAF in transgenic plants. In transgenic plants, many putative chloroplast- or mitochondria-encoded genes were identified as differentially expressed. Our results report comprehensive gene expression changes induced by alterations of seed FA composition and reveal potential targets for further genetic modifications.

Keyword: metabolism

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to lipid and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related disorders.

Keyword: metabolism

causes insulin resistance in granulosa cells via activation of JNK.

Obesity is a worldwide health problem with rising incidence and results in reproductive difficulties. Elevated saturated free fatty acids (FFAs) in obesity can cause insulin resistance (IR) in peripheral tissues. The high intra-follicular saturated FFAs may also account for IR in ovarian granulosa cells (GCs). In the present study, we investigated the relationship between saturated FFAs and IR in GCs by the use of (PA). We demonstrated that the glucose uptake in cultured GCs and lactate accumulation in the culture medium were stimulated by insulin, but the effects of insulin were attenuated by PA treatment. Besides, insulin-induced phosphorylation of Akt was reduced by PA in a dose and time-dependent manner. Furthermore, PA increased phosphorylation of JNK and JNK blockage rescued the phosphorylation of Akt which was down-regulated by PA. These findings highlighted the negative effect of PA on GCs and may partially account for the obesity-related reproductive disorders.

Keyword: metabolism

Beyond the Scavenging of Reactive Oxygen Species (ROS): Direct Effect of Cerium Oxide Nanoparticles in Reducing Fatty Acids Content in an In Vitro Model of Hepatocellular Steatosis.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic accumulation of lipids. Antisteatotic effects of cerium oxide nanoparticles (CeONPs) have recently been shown in animal models of liver disease. However, it is unclear whether the activity of CeONPs is related solely to the decrease in oxidative stress or, in addition, they directly decrease liver fatty accumulation. To address this question, in this work, we used an in vitro model of hepatocellular steatosis, exposing HepG2 cells to oleic and . Cell uptake of CeONPs and their effect on oxidative stress and viability of hepatic cells cultured with HO were also evaluated. Results show that CeONPs were uptaken by HepG2 cells and reduced oxidative stress and improved cell viability. Treatment with oleic and increased lipogenesis and the content of different fatty acids. CeONPs reduced and stearic and most fatty acids consisting of more than 18 carbon atoms. These effects were associated with significant changes in elongase and desaturase activity. In conclusion, CeONPs directly protected HepG2 cells from cell injury in oxidative stress conditions and reduced fatty content in steatotic conditions by inducing specific changes in fatty , thus showing potential in the treatment of NAFLD.

Keyword: metabolism

The Mitochondria-Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

Mitochondria-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive oxygen species (ROS) production during glucose in β cells can be exacerbated under hyperglycaemic conditions such as type 2 diabetes (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: metabolism

Examination of the Catabolic Rates of C-Labeled Fatty Acids Bound to the α and β Positions of Triacylglycerol Using CO Expired from Mice.

Fatty acids in triacylglycerol (TAG) are catabolized after digestion. However, the catabolic rates of several fatty acids bound to the α (sn-1, 3) or β (sn-2) position of TAG have not been thoroughly compared. In this study, the catabolic rates of C-labeled , oleic , linoleic , α-linolenic , eicosapentaenoic (EPA), or docosahexaenoic (DHA) bound to the α and β position of TAG were compared using isotope ratio mass spectrometry. The catabolic rates of the studied fatty acids were evaluated using the ratio of C and C in carbon dioxide expired from mice. The results indicated that , oleic , or α-linolenic bound to the β position was slowly catabolized for a long duration compared to that when bound to the α position. In contrast, EPA bound to the β position was quickly catabolized, and EPA bound to the α position was slowly catabolized for a long time. For linoleic or DHA, no difference in the catabolic rates was detected between the binding positions in TAG. Furthermore, EPA and DHA were less catabolized than the other fatty acids. These results indicate that the catabolic rates of fatty acids are influenced by their binding positions in TAG and that this influence on the catabolic rate differed depending on the fatty species.

Keyword: metabolism

Exogenous addition of alkanoic acids enhanced production of antifungal lipopeptides in Bacillus amyloliquefaciens Pc3.

The bacterium, Bacillus amyloliquefaciens Pc3, was previously isolated from Antarctic seawater and has been found to show antagonistic activity against the fungus, Rhizoctonia solani ACCC 36316, which causes a severe disease known as Sclerotinia sclerotiorum in rapeseed plants. Bacillus lipopeptides had been widely used as biocontrol agents for plant diseases. In this study, we isolated 11 lipopeptide compounds from B. amyloliquefaciens Pc3 culture broth via reversed-phase high-performance liquid chromatography (RP-HPLC) and used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to identify these as iturin A (C, C, C, C), fengycin B (C, C, C, C), and surfactin (C, C, C). We further found that the addition of exogenous alkanoic acids, including myristic , pentadecanoic , , heptadecanoic , octadecanoic , and nonadecanoic , to the bacterial growth media could promote lipopeptide production and enhance the antifungal activities of crude lipopeptide extracts from B. amyloliquefaciens Pc3. In addition, the transcriptional levels of three lipopeptide synthesis genes, ituD, fenA, and srfA-A, and two fatty -related genes, FabI, which encodes enoyl-ACP reductase, and FadB, which encodes enoyl-CoA hydratase, were found to be upregulated in cells grown with exogenous alkanoic acids. Among the six alkanoic acids tested, those with odd carbon chain lengths had a greater effect on lipopeptide production, antifungal activity, and target gene upregulation than those with even carbon chain lengths. These results provide a practical approach for the efficient production of lipopeptides in Bacillus amyloliquefaciens Pc3.

Keyword: metabolism

Palmitate is not an effective fuel for pancreatic islets and amplifies insulin secretion independent of calcium release from endoplasmic reticulum.

The aim of the study was to determine the acute contribution of fuel oxidation in mediating the increase in insulin secretion rate (ISR) in response to fatty acids. Measures of mitochondrial , as reflected by oxygen consumption rate (OCR) and cytochrome c reduction, calcium signaling, and ISR by rat islets were used to evaluate processes stimulated by acute exposure to (PA). The contribution of mitochondrial oxidation of PA was determined in the presence and absence of a blocker of mitochondrial transport of fatty acids (etomoxir) at different glucose concentrations. Subsequent to increasing glucose from 3 to 20 mM, PA caused small increases in OCR and cytosolic calcium (about 20% of the effect of glucose). In contrast, the effect of PA on ISR was almost 3 times that by glucose, suggesting that the of PA is not the dominant mechanism mediating PA\'s effect on ISR. This was further supported by lack of inhibition of PA-stimulated OCR and ISR when blocking entry of PA into mitochondria (with etomoxir), and PA\'s lack of stimulation of reduced cytochrome c in the presence of high glucose. Consistent with the lack of stimulation by PA, an inhibitor of calcium release from the endoplasmic reticulum, but not a blocker of L-type calcium channels, abolished the PA-induced elevation of cytosolic calcium. Notably, ISR was unaffected by thapsigargin showing the dissociation of endoplasmic reticulum calcium release and second phase insulin secretion. In conclusion, stimulation of ISR by PA was mediated by mechanisms largely independent of the oxidation of the fuel.

Keyword: metabolism

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: metabolism

Probing the Global Cellular Responses to Lipotoxicity Caused by Saturated Fatty Acids.

Excessive levels of saturated fatty acids are toxic to cells, although the basis for this lipotoxicity remains incompletely understood. Here, we analyzed the transcriptome, lipidome, and genetic interactions of human leukemia cells exposed to palmitate. Palmitate treatment increased saturated glycerolipids, accompanied by a transcriptional stress response, including upregulation of the endoplasmic reticulum (ER) stress response. A comprehensive genome-wide short hairpin RNA (shRNA) screen identified >350 genes modulating lipotoxicity. Among previously unknown genetic modifiers of lipotoxicity, depletion of RNF213, a putative ubiquitin ligase mutated in Moyamoya vascular disease, protected cells from lipotoxicity. On a broader level, integration of our comprehensive datasets revealed that changes in di-saturated glycerolipids, but not other lipid classes, are central to lipotoxicity in this model. Consistent with this, inhibition of ER-localized glycerol-3-phosphate acyltransferase activity protected from all aspects of lipotoxicity. Identification of genes modulating the response to saturated fatty acids may reveal novel therapeutic strategies for treating diseases linked to lipotoxicity.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 diabetes.

stress-induced low-grade chronic inflammation plays an important role in the development of insulin-resistance and type 2 diabetes (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the obesity-induced stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: metabolism

dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the energy source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive oxygen species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal lipid loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: metabolism

Yeast Smell Like What They Eat: Analysis of Volatile Organic Compounds of in Growth Media Supplemented with Different Lipids.

is part of the human skin microbiota. Its volatile organic compounds (VOCs) possibly contribute to the characteristic odour in humans, as well as to microbiota interaction. The aim of this study was to investigate how the lipid composition of the liquid medium influences the production of VOCs. Growth was performed in four media: (1) mDixon, (2) oleic (OA), (3) oleic + (OA+PA), and (4) (PA). The profiles of the VOCs were characterized by HS-SPME/GC-MS in the exponential and stationary phases. A total number of 61 VOCs was found in , among which alkanes, alcohols, ketones, and furanic compounds were the most abundant. Some compounds previously reported for (γ-dodecalactone, 3-methylbutan-1-ol, and hexan-1-ol) were also found. Through our experiments, using univariate and multivariate unsupervised (Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA)) and supervised (Projection to Latent Structures Discriminant Analysis (PLS-DA)) statistical techniques, we have proven that each tested growth medium stimulates the production of a different volatiles profile in . Carbon dioxide, hexan-1-ol, pentyl acetate, isomer5 of methyldecane, dimethyl sulphide, undec-5-ene, isomer2 of methylundecane, isomer1 of methyldecane, and 2-methyltetrahydrofuran were established as differentiating compounds among treatments by all the techniques. The significance of our findings deserves future research to investigate if certain volatile profiles could be related to the beneficial or pathogenic role of this yeast.

Keyword: metabolism

Comprehensive characterization of nanostructured lipid carriers using laboratory and synchrotron X-ray scattering and diffraction.

The development of lipid nanoparticles requires knowledge on the crystalline structure, polymorphic transitions and lipid-drug interactions. This study aimed at introducing advanced techniques to characterize nanostructured lipid carriers (NLC) comprising , oleic , stabilizer and Domperidone. Crystallinity of single components and mixtures was investigated by laboratory Small Angle X-ray Scattering (SAXS). NLC were studied with laboratory Small and Wide Angle X-ray Scattering (SWAXS). Photon Correlation Spectroscopy and Freeze Fracture Transmission Electron Microscopy were used to monitor particle size, zeta potential and shape. Stability of NLC was investigated using synchrotron X-ray Diffraction (XRD) and SAXS and laboratory SAXS. showed a lamellar structure (polymorph C), which was still present after particle preparation. Spherical 300\u202fnm-sized particles with zeta potential values above -30\u202fmV were obtained and Domperidone was incorporated in its amorphous form. During storage, no differences in synchrotron XRD spectra were seen. However, laboratory SAXS measurements showed a second lamellar structure, identified as polymorph B. Synchrotron SAXS temperature scans confirmed that polymorph B did not affect the morphology of the encapsulated drug or the shape of NLC. These results highlight the unique capabilities of laboratory and synchrotron X-ray Scattering and Diffraction for improved structural characterization of lipid nanoparticles.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolism

A Novel Role for Somatostatin in the Survival of Mouse Pancreatic Beta Cells.

Cross-talk between different pancreatic islet cell types regulates islet function and somatostatin (SST) released from pancreatic delta cells inhibits insulin secretion from pancreatic beta cells. In other tissues SST exhibits both protective and pro-apoptotic properties in a tissue-specific manner, but little is known about the impact of the peptide on beta cell survival. Here we investigate the specific role of SST in the regulation of beta cell survival in response to physiologically relevant inducers of cellular stress including palmitate, cytokines and glucose.Pancreatic MIN6 beta cells and primary mouse islet cells were pre-treated with SST with or without the G signalling inhibitor, pertussis toxin, and exposed to different cellular stress factors. Apoptosis and proliferation were assessed by measurement of caspase 3/7 activity, TUNEL and BrdU incorporation, respectively, and expression of target genes was measured by qPCR.SST partly alleviated upregulation of cellular stress markers (Hspa1a and Ddit3) and beta cell apoptosis in response to factors such as lipotoxicity (palmitate), pro-inflammatory cytokines (IL1β and TNFα) and low glucose levels. This effect was mediated via a G protein-dependent pathway, but did not modify transcriptional upregulation of the specific NFκB-dependent genes, Nos2 and Ccl2, nor was it associated with transcriptional changes in SST receptor expression.Our results suggest an underlying protective effect of SST which modulates the beta cell response to ER stress and apoptosis induced by a range of cellular stressors associated with type 2 diabetes.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: metabolism

Molecular Targets of Fatty Ethanolamides in Asthma.

Asthma is a common allergic pathology of the respiratory tract that requires the study of mechanisms underlying it, due to severe forms of the disease, which are refractory to therapy. The review is devoted to the search for molecular targets of fatty ethanolamides in asthma, in particular palmitoylethanolamide (PEA), which has been successfully used in the treatment of chronic inflammatory and neurodegenerative diseases, in the pathogenesis of which the nervous and immune systems are involved. Recently, the potentially important role of neuro-immune interactions in the development of allergic reactions has been established. Many of the clinical symptoms accompanying allergic airway inflammation are the result of the activation of neurons in the airways, so the attention of researchers is currently focused on neuro-immune interactions, which can play an important role in asthma pathophysiology. A growing number of scientific works confirm that the key molecule in the implementation of these inter-systemic interactions is nerve growth factor (NGF). In addition to its classic role in nervous system physiology, NGF is considered as an important factor associated with the pathogenesis of allergic diseases, particularly asthma, by regulating of mast cell differentiation. In this regard, NGF can be one of the targets of PEA in asthma therapy. PEA has a biological effect on the nervous system, and affects the activation and the degranulation of mast cells.

Keyword: metabolism

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy . However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: metabolism

fingerprinting unveils quinoa oil as a source of bioactive phytochemicals.

Quinoa is a pseudo-cereal with high phytochemical contents with proven biological and nutritional significance. In recent decades several breeding programmes have introduced new and traditional quinoa varieties to North America and other non-traditional quinoa regions, raising questions regarding variability in their secondary metabolite profiles. In this work, we have fingerprinted 28 quinoa varieties cultivated in Washington State, focussing on the poorly investigated oil fraction. We found variability in both phenolics and carotenoid contents, which is reflected in different antioxidant capacities, as measured by FRAP and DPPH. Fatty profiles show significant differences in and long-chain fatty acids. Finally, conspicuous amounts of phytosterols and squalene were found. Through factor analysis, we classified the quinoa varieties into two groups: a first comprising varieties with higher phytochemical and PUFA contents and a second group, with higher linolenic and long-chain fatty contents. Both groups may be suitable for potential food applications.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Mangiferin Improved Palmitate-Induced-Insulin Resistance by Promoting Free Fatty in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved Insulin Resistance.

Elevated free fatty (FFA) is a key risk factor for insulin resistance (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high-fat diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA in HepG2 and C2C12 cells. The model was used to quantify PA-induced lipid accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased insulin-stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: metabolism

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: metabolism

A worldwide reliable indicator to differentiate wild vs. farmed Penaeid shrimps based on 207 fatty profiles.

Shrimps and prawns are especially subject to food fraud, which has consequences not only on the economy but also represents a potential risk for public health. Fatty acids (FA) of Penaeid shrimps have been largely explored in the literature, and although they are unable to discriminate shrimps geographical origin or species, they might provide an interesting tool to distinguish their production method (wild vs. farmed). The present study is based on a literature compilation of Penaeid shrimp FA profiles encompassing all continents and 28 species. It reveals that the ratio of FA 18:2ω6\u202f+\u202fFA 18:3ω3 / FA 16:1ω7 can differentiate wild vs. farmed Penaeid shrimps with 100% accuracy within the 207 FA profiles of the dataset considered. Assuming a normal distribution of the dataset, 94.4% of the farmed shrimps population is expected to exhibit a ratio above 2.92, and 99.7% of the wild shrimps population is expected to fall below 2.92.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Palm Oil and Beta-palmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition.

Palm oil (PO) is used in infant formulas in order to achieve (PA) levels similar to those in human milk. PA in PO is esterified predominantly at the SN-1,3 position of triacylglycerol (TAG), and infant formulas are now available in which a greater proportion of PA is in the SN-2 position (typical configuration in human milk). As there are some concerns about the use of PO, we aimed to review literature on health effects of PO and SN-2-palmitate in infant formulas.PubMed and Cochrane Database of Systematic Reviews were systematically searched for relevant studies on possible beneficial effects or harms of either PO or SN-2-palmitate in infant formula on various health outcomes.We identified 12 relevant studies using PO and 21 studies using SN-2-palmitate. Published studies have variable methodology, subject characteristics, and some are underpowered for the key outcomes. PO is associated with harder stools and SN-2-palmitate use may lead to softer stool consistency. Bone effects seem to be short-lasting. For some outcomes (infant colic, faecal microbiota, lipid ), the number of studies is very limited and summary evidence inconclusive. Growth of infants is not influenced. There are no studies published on the effect on markers of later diseases.There is insufficient evidence to suggest that PO should be avoided as a source of fat in infant formulas for health reasons. Inclusion of high SN-2-palmitate fat blend in infant formulas may have short-term effects on stool consistency but cannot be considered essential.

Keyword: metabolism

Comparison of monoculture and mixed culture (Scenedesmus obliquus and wild algae) for C, N, and P removal and lipid production.

This study compared the efficiency of nutrient removal and lipid accumulation by a monoculture of Scenedesmus obliquus and mixed cultures of microalgae. The highest removal efficiencies of ammonium (99.2%), phosphate (91.2%), and total organic carbon (83.6%) occurred in the monoculture. All the mixed cultures were dominated by S. quadricauda; in some mixed cultures, the proportions of Chlamydomonas reinhardtii and C. microsphaera reached >\u200920%. The lipid content and lipid production in the monoculture were 15.9% and 52.3\xa0mg\xa0kg, respectively, significantly higher than those in all the mixed cultures of microalgae. In all the mixed cultures, the proportion of was >\u200950%. The results suggest that the monoculture had advantages over the mixed culture of microalgae in terms of nutrient removal and lipid production.

Keyword: metabolism

Early and reversible changes to the hippocampal proteome in mice on a high-fat diet.

The rise in global obesity makes it crucial to understand how diet drives obesity-related health conditions, such as premature cognitive decline and Alzheimer\'s disease (AD). In AD hippocampal-dependent episodic memory is one of the first types of memory to be impaired. Previous studies have shown that in mice fed a high-fat diet (HFD) episodic memory is rapidly but reversibly impaired.In this study we use hippocampal proteomics to investigate the effects of HFD in the hippocampus. Mice were fed either a low-fat diet (LFD) or HFD containing either 10% or 60% (Kcal) from fat for 3\u2009days, 1\u2009week or 2\u2009weeks. One group of mice were fed the HFD for 1\u2009week and then returned to the LFD for a further week. Primary hippocampal cultures were challenged with (PA), the most common long-chain saturated FA in the Western diet, and with the anti-inflammatory, n-3 polyunsaturated FA, docosahexaenoic (DHA), or a combination of the two to ascertain effects of these fatty acids on dendritic structure.HFD-induced changes occur in hippocampal proteins involved in , inflammation, cell stress, cell signalling, and the cytoskeleton after 3\u2009days, 1\u2009week and 2\u2009weeks of HFD. Replacement of the HFD after 1\u2009week by a low-fat diet (LFD) for a further week resulted in partial recovery of the hippocampal proteome. Microtubule-associated protein 2 (MAP2), one of the earliest proteins changed, was used to investigate the impact of fatty acids (FAs) on hippocampal neuronal morphology. PA challenge resulted in shorter and less arborised dendrites while DHA had no effect when applied alone but counteracted the effects of PA when FAs were used in combination. Dendritic morphology recovered when PA was removed from the cell culture media.This study provides evidence for the rapid and reversible effects of diet on the hippocampal proteome and the impact of PA and DHA on dendritic structure.

Keyword: metabolism

Green preparation of seaweed-based silver nano-liquid for cotton pathogenic fungi management.

Silver nanoparticles (Ag NPs) were synthesised using the crude ethyl acetate extracts of and evaluated their bioefficacy against two crop-damaging pathogens. The sets of lattice planes in the XRD spectrum for the Ag NPs were indexed to the 111, 200, 220 and 311 orientations and support the crystalline nature of the Ag NPs. The 3414 and 2968\u2005cm peaks were observed in crude algal thallus extract and they were characteristic of terpenoids. Further, a peak at 1389\u2005cm was observed as fatty acids. The marine macroalgae terpenoids and acted as reducing agent and stabiliser, respectively. The size (3 and 50\u2005nm) and shape (spherical) of Ag NPs were recorded. The energy-dispersive X-ray spectroscopy analysis exemplified the presence of silver in its elemental nature. Moreover, Ag NPs were effective against two cotton phytopathogens namely f.sp. (FOV) and pv. malvacearum (XAM). The minimum inhibitory concentration was found to be 80.0 and 43.33\u2005μg ml against FOV and XAM, respectively. Results confirmed the anti-microbial activity of green nanoparticles against select pathogens and suggest their possible usage in developing antifungal agents for controlling destructive pathogens in a cotton agroecosystem.

Keyword: metabolism

Analysis of Fatty Esters of Hydroxyl Fatty in Selected Plant Food.

syndrome, characterized by obesity, low-grade inflammation, insulin resistance, hyperglycemia, dyslipidemia and hypertension, is a major risk factor for cardiovascular mortality. Preclinical studies on recently discovered classes of lipids - fatty esters of hydroxy fatty acids (FAHFA) have revealed their anti-inflammatory and insulin-sensitizing potential. The FAHFA levels are significantly decreased in insulin-resistant individuals, their application exhibited anti-inflammatory effects and restoring the glucose-insulin homeostasis. The aim of our research was to analyze the overall FAHFA composition in a common diet, as only a partial FAHFA composition has been revealed so far (only the PAHSA subclass was analyzed in a few foods). A new approach to the FAHFAs analysis includes nano-LC and post-column modifier followed by negative ion mass spectrometry, in order to obtain maximum sensitivity. Analysis of different foods - oat (whole grain, coarse flakes and fine flakes), apple, clementine, lemon, strawberry, blueberry, mango, kiwi, avocado, pineapple, banana, onion, garlic, cherry tomato, carrot, parsley root, pepper and radish - exhibited wide inter-food variation in the FAHFA profiles. Sixteen analyzed FAHFAs (, oleic, palmitoleic and stearic hydroxy-esters) showed microgram to low nanogram levels (0.165\xa0ng/g - 32\xa0μg/g FW), with the highest abundancy in oat, clementine, garlic and pineapple. Stearic hydroxy stearic (SAHSA) was the most abundant FAHFA, especially in the food with antioxidative, anti-inflammatory and beneficial effects. In contrary, the PAHSA - previously proven to have the strongest antihyperglycemic and insulin-sensitizing effects, was not present in some foods (radish, avocado, mango, lemon, cherry tomato, kiwi). Our study proves the importance of overall FAHFA analysis in food (especially in a functional food), because of their potential benefits and possible future incorporation in special diets.

Keyword: metabolism

Fatty profiles of the European migratory common noctule bat (Nyctalus noctula).

In animals, fatty acids (FA) are essential as structural components in membranes and for energy storage in adipocytes. Here, we studied the relative proportions of FA in a mammal with extreme changes in rates. Common noctule bats (Nyctalus noctula) switch from energetically demanding long-distance migration at high rates to regular torpor with extremely low rates. We found that composition of FA categories differed between adipose tissue types (white adipose tissue (WAT) vs brown adipose tissue (BAT)) and muscle tissue types (skeletal vs heart), but not between sexes. We found oleic to be the most abundant FA in all studied tissues. Concentrations of polyunsaturated FA (PUFA) were not always higher in muscular tissue compared with adipocyte tissue, even though high concentrations of PUFA are considered beneficial for low body temperatures in torpor. In all tissues, we observed a high content in monounsaturated fatty acids (MUFA), possibly to compensate for a low PUFA content in the diet. Ratios of ω6/ω3 were lower in the heart than in skeletal muscles of common noctules. Three FA (, oleic, and linoleic ) accounted for about 70% of the FA in adipose tissue, which is similar to proportions observed in migrating birds, yet migrating birds generally have a higher PUFA content in muscle and adipose tissues than bats. Bats seem to contrast with other mammals in having a high MUFA content in all tissues. We conclude that FA profiles of bats differ largely from those of most cursorial mammals and instead are-with the exception of MUFA-similar to those of migrating birds.

Keyword: metabolism

Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic fatty liver disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high-fat diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of fatty liver disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: metabolism

Dietary habits affect fatty composition of visceral adipose tissue in subjects with colorectal cancer or obesity.

Aim of this study was to identify a possible relationship among dietary fatty acids (FA) intake, FA adipose tissue (AT) profile and cancer condition in lean vs obese subjects affected or not by colorectal cancer (CRC). Actually, inadequate dietary habits together with physical inactivity are primary determinants of obesity and cancer risk. Changes in lipid play a crucial role in different types of cancer and key enzymes involved in lipid- , such as stearoyl-coA-desaturase 1 (SCD-1), are differentially expressed in normal and cancer tissues.Food frequency questionnaires (FFQ) were analyzed by Winfood software. FA were assessed by gas-liquid chromatography in visceral AT samples. Estimated desaturase activities were calculated as precursor FA/product FA ratio. Desaturase gene expressions were evaluated by RT-qPCR.Lean and obese CRC subjects showed inadequate dietary habits. In particular, lean CRC subjects showed increase in the intake of saturated FA, specifically (p\u2009=\u20090.0042) and stearic (p\u2009=\u20090.0091), and a corresponding reduction of monounsaturated FA consumption, in particular oleic (p\u2009=\u20090.002) with respect to lean without CRC. Estimated SCD-1 activity in AT was increased in all the groups vs lean without CRC (pANOVA\u2009=\u20090.029).Unhealthy eating habits, characterizing obese and CRC subjects, may influence the visceral AT profile and contribute to the alteration of the . The quality of the diet, other than the quantity, can have a main role in the establishment of inflammatory microenvironment and in changes favouring CRC.

Keyword: metabolism

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid . We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: metabolism

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK .

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose , such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in lipid , including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated .

Keyword: metabolism

Telmisartan protects against high glucose/high lipid-induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: metabolism

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the of fatty acids (FA) and only in the second turn in glucose ; regulation of FA in cells started millions of years earlier than that of glucose . Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: metabolism

Relationship between acyl-lipid and sterol metabolisms in diatoms.

Diatoms are a phylum of unicellular photosynthetic eukaryotes living in oceans and fresh waters, characterized by the complexity of their plastid, resulting from a secondary endosymbiosis event. In the model diatom Phaeodactylum tricornutum, fatty acids (FAs) are synthesized from acetyl-CoA in the stroma of the plastid, producing . FAs are elongated and desaturated to form very-long chain polyunsaturated fatty acids (VLC-PUFAs) in domains of the endomembrane system that need to be identified. Synthesis of VLC-PUFAs is coupled with their import to the core of the plastid via the so-called "omega" pathway. The biosynthesis of sterols in diatoms is likely to be localized in the endoplasmic reticulum as well as using precursors deriving from the mevalonate pathway, using acetyl-CoA as initial substrate. These modules can be characterized functionally by genetic analyzes or chemical treatments with appropriate inhibitors. Some \' modules\' are characterized by a very low level of intermediates. Since some chemical treatments or genetic perturbation of lipid induce the accumulation of these intermediates, channeling processes are possibly involved, suggesting that protein-protein interactions might occur between enzymes within large size complexes or metabolons. At the junction of these modules, intermediates might therefore play dramatic roles in directing carbon fluxes from one direction to another. Here, acetyl-CoA seems determinant in the balance between TAGs and sterols. Future lines of research and potential utilization for biotechnological applications are discussed.Published by Elsevier B.V.

Keyword: metabolism

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and lipid .

Fibroblast growth factor 21 (FGF21) is known as a potent regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory whereby FGF21 mediates islet lipid in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high-fat diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced lipid accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced lipid deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell lipid accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: metabolism

Factors associated with the content of mammary-synthesized fatty acids in milk fat: A meta-analysis.

Consumption of specific fatty acids (FA) that are synthesized in the mammary gland, namely de novo FA, has implications for human health. The objective of the present meta-analysis was to study the associations between milk fat content of de novo FA, with (1) diet composition, and (2) milk production and composition. Milk FA data from 96 peer-reviewed studies published between 1990 and 2016 that included 324 treatment means from 83 bovine experiments, 36 treatment means from 12 caprine experiments, and 40 treatment means from 12 ovine experiments were used in this analysis. Individual species models including the fixed effect of experiment were fitted using multiple regression to explain milk content of de novo FA as a function of diet composition and milk production and composition variables. We also evaluated replacing the effect of the experiment by the effect of the experiment nested in the laboratory at which the research had been conducted, and the effect of the laboratory. Butyric content in milk fat was positively but weakly related to dietary ether extract in does and ewes. Lauric, myristic, and contents in milk fat were negatively related to dietary ether extract in does and to a somewhat lesser extent in cows and ewes. The results confirm that the inclusion of lipids in the diet may not only affect the availability of preformed FA but also the profile of FA synthesized de novo in the mammary gland. Most of the variation in all prediction models was explained by the experiment or by the laboratory if the latter was included in the model. The ample variation in analytical methods reported by the different research groups suggests that differences in analytical protocols might explain a substantial proportion of the variation in de novo FA profile. A main conclusion of this study is the potential influence of differences in analytical procedures to explain the variation in de novo FA profile. Standardization of methods of FA analysis to improve reproducibility seems to be an aspect of importance to this area of research.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

Flaxseed oil ameliorated high-fat-diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts.

α-Linolenic (ALA) is a plant-derived omega-3 unsaturated fatty that is rich in flaxseed oil (FO). The effect of FO on bone health is controversial. This study aims to evaluate the effect of FO on bone damage induced by a high-fat diet (HFD) and to explore the possible mechanism.Male Sprague-Dawley rats were fed a normal control diet (NC, 10% fat), FO diet (NY, 10% fat), HFD (60% fat), or HFD containing 10% FO (HY, 60% fat) for 22\u2009weeks. Micro CT and three-point bending tests were conducted to evaluate bone microstructure and biomechanics. Serum was collected for the detection of ALP, P1NP, and CTX-1. Rat primary osteoblasts (OBs) were treated with different concentrations of ALA with or without (PA) treatment. The ALP activity, osteogenic-related gene and protein expression were measured.Rats in the HFD group displayed decreased biomechanical properties, such as maximum load, maximum fracture load, ultimate tensile strength, stiffness, energy absorption, and elastic modulus, compared with the NC group ( \xa0<\u20090.05). However, HY attenuated the HFD-induced decreases in bone biomechanical properties, including maximum load, maximum fracture load, and ultimate tensile strength (\u2009<\u20090.05). Trabecular bone markers such as trabecular volume bone mineral density (Tb. vBMD trabecular bone volume/total volume (Tb. BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) were decreased, trabecular separation (Tb. Sp) and the structure model index (SMI) were increased in the HFD group compared with the NC group, and all parameters were remarkably improved in the HY group compared to the HFD group (p\u2009<\u20090.05). However, cortical bone markers such as cortical volume bone mineral density (Ct. vBMD), cortical bone volume/total volume (Ct. BV/TV) and cortical bone thickness (Ct. Th) were not significantly different among all groups. Moreover, the serum bone formation markers ALP and P1NP were higher and the bone resorption marker CTX-1 was lower in the HY group compared with levels in the HFD group. Compared with the NC group, the NY group had no difference in the above indicators. In rat primary OBs, PA treatment significantly decreased ALP activity and osteogenic gene and protein (β-catenin, RUNX2, and osterix) expression, and ALA dose-dependently restored the inhibition induced by PA.FO might be a potential therapeutic agent for HFD-induced bone loss, most likely by promoting osteogenesis.© The Author(s). 2019.

Keyword: metabolism

Anthocyanins ameliorate palmitate-induced inflammation and insulin resistance in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade inflammation leading to insulin resistance. Adipocyte differentiation inhibitors are expected to be effective in preventing obesity and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, inflammation, and insulin resistance in 3T3-L1 adipocytes. ACN extract pretreatment reduces lipid accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces inflammation with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces insulin resistance by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce insulin pathway with levels higher than insulin control cells, supporting an insulin sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against obesity comorbidities, due to their protective effects against inflammation/insulin resistance in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and insulin resistance conditions related to obesity.© 2019 John Wiley & Sons, Ltd.

Keyword: metabolism

Protocatechuic -Ameliorated Endothelial Oxidative Stress through Regulating Acetylation Level via CD36/AMPK Pathway.

As one of the main metabolites of anthocyanin, protocatechuic (PCA) possesses strong antioxidant activity. In the present study, we explored the capacity of PCA on the alleviation of endothelial oxidative stress and investigated the underlying mechanisms using RNA sequencing (RNA-Seq). In comparison with (PA)-treated cells, PCA (100 μM) significantly decreased the generations of 3-nitrotyrosine (3-NT) and 8-hydroxydeoxyguanosine (8-OHdG) (0.82 ± 0.01 vs 1.16 ± 0.05 and 0.80 ± 0.01 vs 1.48 ± 0.15, respectively, p < 0.01), two biomarkers of oxidative damage, and restored the levels of nitric oxide (NO) (0.97 ± 0.04 vs 0.54 ± 0.02, p < 0.01) and mitochondrial membrane potential (MMP) (0.96 ± 0.03 vs 0.86 ± 0.02, p < 0.01) in human umbilical vein endothelial cells (HUVECs). PCA also obviously reduced the level of reactive oxygen species (ROS) (0.86 ± 0.15 vs 2.67 ± 0.09, p < 0.01) in aorta from high-fat diet (HFD)-fed mice. RNA-Seq and Western blot analysis indicated that PCA markedly reduced the expression of cluster of differentiation 36 (CD36), a membrane fatty transporter, and reduced the generations of adenosine triphosphate (ATP) and acetyl coenzyme A (Ac-CoA). These effects of PCA were associated with decreased level of acetylated-lysine and restored the activity of manganese-dependent superoxide dismutase (MnSOD) through reducing the generation of Ac-CoA or activating Sirt1 and Sirt3 via a CD36/AMP-kinase (AMPK) dependent pathway.

Keyword: metabolism

[Screening of serum susceptibility biomarkers in patients with mild liver function abnormalities based on non-targeted metabolomics].

In the present study,non-targeted metabolomics technique was used to screen potentially susceptibility biomarkers in patients with mild liver function abnormalities during long-term use of Chinese herbal compound. According to the inclusion and exclusion criteria,we collected 7 cases of patients with abnormal liver function during the period of complete taking Chinese herbal medicine( 60 days),and 18 cases of patients with normal liver function in re-examination from the reproductive medicine center in our hospital. Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF/MS~E) technique combined with Progenesis QI software was used to analyze the differential biomarkers in serum of patients with wild liver function abnormalities and normal liver function. 11 potential biomarkers such as bilirubin,pantothenic ,hippuric ,sphingomyelin,,and oleic were tentatively identified. disorders in patients with herbal-induced mild liver abnormality were mainly related to two : pantothenic and coenzyme A biosynthesis and linoleic . It could provide a reference for the early warning of mild liver function abnormalities of patients that may be caused by long-term use of Chinese medicine compound in clinical application,and will lay a foundation for further understanding the endogenous substance changes in different levels of liver injury.

Keyword: metabolism

Chokeberry Juice Containing Polyphenols Does Not Affect Cholesterol or Blood Pressure but Modifies the Composition of Plasma Phospholipids Fatty Acids in Individuals at Cardiovascular Risk.

Chokeberry polyphenols have been suggested to reduce cholesterol and blood pressure and thus protect against cardiovascular diseases (CVD), but the evidence in humans is limited and inconsistent. This randomized double-blinded three-parallel groups trial investigated the changes in various anthropometric and clinical biomarkers, and in plasma phospholipids fatty acids (PPFA) in volunteers at cardiovascular risk after a four-week intervention with 100 mL/day of (1) chokeberry juice with a high-dose of polyphenols (1177.11 mg gallic equivalents, GAE); (2) chokeberry juice with a low-dose of polyphenols (294.28 mg GAE) and; (3) a nutritionally matched polyphenol-free placebo drink. Our results indicate that the intake of chokeberry juice containing either the low or the high dose of polyphenols cannot be linked with a reduction in total- and low-density lipoprotein (LDL)cholesterol or in systolic (SBP) and diastolic (DBP) blood pressure in comparison with the consumption of the placebo drink. However, we found evidence of moderate changes in the PPFA, i.e., increased saturated fatty acids (SFA), mostly , and reduced n-6 polyunsaturated fatty acids (PUFA), principally linoleic (LA) with the intake of chokeberry against the placebo. These effects may be associated with the polyphenols but we could not differentiate a clear dose-response effect. Further research is still needed to elucidate the contribution of the polyphenolic fraction to the potential cardiovascular effects of the chokeberry and to build up the evidence of its potential benefit via the modulation of PPFA composition.

Keyword: metabolism

Fatty in the host and commensal bacteria for the control of intestinal immune responses and diseases.

Intestinal tissue has a specialized immune system that exhibits an exquisite balance between active and suppressive responses important for the maintenance of health. Intestinal immunity is functionally affected by both diet and gut commensal bacteria. Here, we review the effects of fatty acids on the regulation of intestinal immunity and immunological diseases, revealing that dietary fatty acids and their metabolites play an important role in the regulation of allergy, inflammation, and immunosurveillance in the intestine. Several lines of evidence have revealed that some dietary fatty acids are converted to biologically active metabolites by enzymes not only in the host but also in the commensal bacteria. Thus, biological interaction between diet and commensal bacteria could form the basis of a new era in the control of host immunity and its associated diseases.

Keyword: metabolism

Saturated fatty stimulates production of extracellular vesicles by renal tubular epithelial cells.

Lipotoxicity, an accumulation of intracellular lipid metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of disease. , a predominant lipid derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated (PA), and BSA-conjugated oleic (OA) for 24-48\xa0h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30-150\xa0nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of lipid-induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of lipid-induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keyword: metabolism

Rice bran oil ameliorates inflammatory responses by enhancing mitochondrial respiration in murine macrophages.

Previous studies have revealed the anti-inflammatory properties of rice bran oil (RBO), but the detailed mechanisms are poorly understood. Recent studies on the molecular/cellular anti-inflammatory mechanisms of dietary components have demonstrated that mitochondrial respiration plays a key role in macrophage functioning. Since dietary lipids are major substrates for mitochondrial respiration through β-oxidation, the current study examined whether RBO regulates inflammatory responses by modulating mitochondrial energy . Palm oil (PO), enriched with which are known to be effectively taken up by cells and used for oxidative phosphorylation, served as a positive control. In the in vitro model of LPS-stimulated RAW 264.7 murine cells, the levels of pro-inflammatory cytokines (IL-6 and TNF-α) in the culture supernatant were significantly reduced by RBO treatment. In contrast, secretion of the anti-inflammatory cytokine IL-10 was upregulated by RBO. Transcription of genes encoding inflammatory mediator molecules (COX-2 and iNOS) and expression of activation markers (CD80, CD86, and MHC-II) in LPS-stimulated RAW 264.7 cells were suppressed by RBO. Mitochondrial respiration (as assessed by an extracellular flux analyzer) increased upon RBO treatment, as the basal respiration, maximal respiration, ATP production, and spare respiratory capacity were upregulated. In an in vivo study, C57BL/6 mice were fed a negative control diet containing corn oil (CO), PO, or RBO for 4 weeks, and bone marrow-derived macrophages (BMDM) were isolated from their tibias and femurs. In pro-inflammatory M1-polarized BMDM (M1-BMDM), the RBO-induced suppression of IL-6 and TNF-α was recapitulated in vivo. Mitochondrial respiration in M1-BMDM also increased following the RBO intervention and the PO control treatment as compared to CO fed negative control. Overall, the current study for the first time demonstrates that RBO regulates inflammatory responses in murine macrophages by upregulating mitochondrial respiration. Further clinical studies are required to validate the animal study.

Keyword: metabolism

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced insulin resistance that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty , insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

GC-MS Metabolomics Reveals Distinct Profiles of Low- and High-Grade Bladder Cancer Cultured Cells.

Previous studies have shown that metabolomics can be a useful tool to better understand the mechanisms of carcinogenesis; however, alterations in biochemical that lead to bladder cancer (BC) development have hitherto not been fully investigated. In this study, gas chromatography-mass spectrometry (GC-MS)-based metabolomics was applied to unveil the alterations between low-grade and high-grade BC cultured cell lines. Multivariable analysis revealed a panel of metabolites responsible for the separation between the two tumorigenic cell lines. Significantly lower levels of fatty acids, including myristic, , and palmitoleic acids, were found in high-grade versus low-grade BC cells. Furthermore, significantly altered levels of some amino acids were observed between low- and high-grade BC, namely glycine, leucine, methionine, valine, and aspartic . This study successfully demonstrated the potential of metabolomic analysis to discriminate BC cells according to tumor aggressiveness. Moreover, these findings suggest that bladder tumorigenic cell lines of different grades disclose distinct profiles, mainly affecting fatty biosynthesis and amino to compensate for higher energetic needs.

Keyword: metabolism

Palmitate and Stearate are Increased in the Plasma in a 6-OHDA Model of Parkinson\'s Disease.

Parkinson\'s disease (PD) is the second most common neurodegenerative disorder, without any widely available curative therapy. Metabolomics is a powerful tool which can be used to identify unexpected pathway-related disease progression and pathophysiological mechanisms. In this study, metabolomics in brain, plasma and liver was investigated in an experimental PD model, to discover small molecules that are associated with dopaminergic cell loss.Sprague Dawley (SD) rats were injected unilaterally with 6-hydroxydopamine (6-OHDA) or saline for the vehicle control group into the medial forebrain bundle (MFB) to induce loss of dopaminergic neurons in the substantia nigra pars compacta. Plasma, midbrain and liver samples were collected for profiling. Multivariate and univariate analyses revealed metabolites that were altered in the PD group.In plasma, ( = 3.72 × 10, FC = 1.81) and stearic ( = 3.84 × 10, FC = 2.15), were found to be increased in the PD group. ( = 3.5 × 10) and stearic ( = 2.7 × 10) correlated with test scores indicative of motor dysfunction. Monopalmitin ( = 4.8 × 10, FC = -11.7), monostearin ( = 3.72 × 10, FC = -15.1) and myo-inositol ( = 3.81 × 10, FC = -3.32), were reduced in the midbrain. The liver did not have altered levels of these molecules.Our results show that saturated free fatty acids, their monoglycerides and myo-inositol in the midbrain and enteric circulation are associated with 6-OHDA-induced PD pathology.

Keyword: metabolism

Comparison of Fatty Profiles in a Group of Female Patients with Chronic Kidney Diseases (CKD) and Syndrome (MetS)⁻Similar Trends of Changes, Different Pathophysiology.

Fatty (FA) profiles in the plasma of patients with syndrome and chronic kidney disease (CKD) seem to be identical despite their different etiology (dietary mistakes vs. cachexia). The aim of this study was to compare both profiles and to highlight the differences that could influence the improvement of the treatment of patients in both groups. The study involved 73 women, including 24 patients with chronic kidney disease treated with haemodialysis, 19 patients with syndrome (MetS), and 30 healthy women in the control group. A total of 35 fatty acids and derivatives were identified and quantified by gas chromatography. Intensified elongation processes from C10:0 to C16:0 were noted in both groups (more intense in MetS), as well as an increased synthesis of arachidonic (C20:4n6), which was more intense in CKD. Significant correlations of oleic (C18:1n9), gamma linoleic (C18:3n6), and docosatetraenoate (C22:4n6) with parameters of CKD patients were observed. In the MetS group, auxiliary of oleic were activated, which simultaneously inhibited the synthesis of eicosapentanoic (EPA) and docosahexaenoic (DHA) from alpha lipoic (ALA). On the other hand, in the group of female patients with CKD, the synthesis of EPA and DHA was intensified. Activation of the synthesis of oleic (C18: 1n9 ct) and trans-vaccinic (C18:1) is a protective mechanism in kidney diseases and especially in MetS due to the increased concentration of saturated fatty (SFA) in plasma. The cause of the increased amount of all FAs in plasma in the CKD group, especially in the case of (C16:0) and derivatives stearic (C18:0) acids, may be the decomposition of adipose tissue and the progressing devastation of the organism, whereas, in the MetS group, dietary intake seems to be the main reason for the increase in SFA. Moreover, in MetS, auxiliary are activated for oleic , which cause the simultaneous inhibition of EPA and DHA synthesis from ALA, whereas, in the CKD group, we observe an increased synthesis of EPA and DHA. The higher increase of nervonic (C24:1) in CKD suggests a higher degree of demyelination and loss of axons.

Keyword: metabolism

Sodium Orthovanadate Changes Fatty Composition and Increased Expression of Stearoyl-Coenzyme A Desaturase in THP-1 Macrophages.

Vanadium compounds are promising antidiabetic agents. In addition to regulating glucose , they also alter lipid . Due to the clear association between diabetes and atherosclerosis, the purpose of the present study was to assess the effect of sodium orthovanadate on the amount of individual fatty acids and the expression of stearoyl-coenzyme A desaturase (SCD or Δ-desaturase), Δ-desaturase, and Δ-desaturase in macrophages. THP-1 macrophages differentiated with phorbol 12-myristate 13-acetate (PMA) were incubated in vitro for 48\xa0h with 1\xa0μM or 10\xa0μM sodium orthovanadate (NaVO). The estimation of fatty composition was performed by gas chromatography. Expressions of the genes SCD, fatty desaturase 1 (FADS1), and fatty desaturase 2 (FADS2) were tested by qRT-PCR. Sodium orthovanadate in THP-1 macrophages increased the amount of saturated fatty acids (SFA) such as and stearic , as well as monounsaturated fatty acids (MUFA)-oleic and palmitoleic . Sodium orthovanadate caused an upregulation of SCD expression. Sodium orthovanadate at the given concentrations did not affect the amount of polyunsaturated fatty acids (PUFA) such as linoleic , arachidonic , eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA). In conclusion, sodium orthovanadate changed SFA and MUFA composition in THP-1 macrophages and increased expression of SCD. Sodium orthovanadate did not affect the amount of any PUFA. This was associated with a lack of influence on the expression of FADS1 and FADS2.

Keyword: metabolism

Effect of dietary n-3 polyunsaturated fatty supplementation and post-insemination plane of nutrition on systemic concentrations of analytes, progesterone, hepatic gene expression and embryo development and survival in beef heifers.

Nutrition, and particularly dietary energy intake, plays a fundamental role in reproductive function in cattle. There is some evidence that supplemental omega-3 dietary polyunsaturated fatty acids (n-3 PUFA) can exert positive effects on fertility. The objectives of this study were to evaluate the effect of dietary n-3 PUFA supplementation, post-insemination energy plane of nutrition and their interaction on embryo survival in cattle. Crossbred beef heifers (n\u202f=\u202f185) were individually offered barley straw ad libitum and 6\u202fkg DM of concentrate supplemented with either a rumen-protected source of saturated fatty (; control, CON) or a partially rumen-protected n-3 PUFA-enriched supplement (n-3 PUFA). Estrous was synchronised using two injections of PG administered at 11-d intervals and following artificial insemination (AI\u202f=\u202fDay 0) 179 heifers exhibiting oestrus were inseminated and assigned to one of two dietary treatments: (i) remain on their pre-insemination high dietary plane of nutrition (High) or (ii) restricted to 0.6\xa0× estimated maintenance energy requirements (Low) in a 2\u202f×\u202f2 factorial design. The heifers were then maintained on their assigned diets until slaughter and embryo recovery on Day 16 (n\u202f=\u202f92) or pregnancy diagnosis by ultrasound scanning at Day 30 post-AI (n\u202f=\u202f87). Plasma concentrations of fatty acids, metabolites, insulin, progesterone (P4) and insulin-like growth factor 1 (IGF-1) were measured at appropriate intervals. Hepatic expression of mRNA for aldo-keto reductase (AKR1C), cytochrome P450 2C (CYP 2C) and cytochrome P450 3A (CYP 3A) was examined. The n-3 PUFA supplementation increased plasma n-3 PUFA concentration (P\u202f<\u202f0.05) and reduced n-6: n-3 PUFA ratio (P\u202f<\u202f0.05). Plasma IGF-1 was higher for n-3 PUFA relative to the CON (P\u202f<\u202f0.05) and for High compared with Low plane of nutrition post-AI (P\u202f<\u202f0.05) groups. A low plane of nutrition post-AI increased plasma concentrations of progesterone from Days 7-16 after insemination (P\u202f<\u202f0.001) but reduced embryo length (P\u202f<\u202f0.001). Supplementation with n-3 PUFA reduced and tended to reduce hepatic expression of CYP2C (P\u202f=\u202f0.01) and CYP3A (P\u202f=\u202f0.08), respectively. However, while dietary n-3 PUFA supplementation and an abrupt reduction in nutrient status following insemination elevated plasma concentrations of n-3 PUFA and mid and late phase P4, respectively, there was no effect of either PUFA supplementation or post-insemination plane of nutrition on embryo survival.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

[Lipolysis in very low density lipoproteins - locus minoris resistentiae - in the pathogenesis of hypertriglyceridemia. Positive effects of diet, polyenic fatty acids, statins and fibrates.]

Inhibition of hydrolysis of and oleic triglycedires (TG) in very low density lipoproteins (VLDL), slow formation of active apoВ-100 conformation, blockade of апоЕ/В-100 ligand formation in VLDL and their reduced uptake by insulin-dependent cells cause hypertriglyceridemia (HTG). and oleic VLDL (>80% total VLDL) are not converted in low density lipoproteins (LDL). Atherosclerosis is not an alimentary deficiency of polyenic fatty acids (PFA), but results from low in vivo bioavailability of PFA in LDL against the background of high dietary FA and LDL. Plasma PFA content and cellular PFA deficiency are as high as LDL cholesterol (CL). Primary prevention of atherosclerosis should be based on a decrease in dietary content of saturated FA, trans FA and a moderate increase in PFA. It seems highly unlikely that the xeobiotics statins, fibrates and probucol produce pleiotropic biological effects in vivo. These effects are brought about by phylogenetically early humoral mediators eicosanoids: prostacyclins, prostaglandins, thromboxanes, leukotrienes, and resolvins. It is reasonable to suggest that all preparations which act according to the same algorithm activate TG hydrolysis in VLDL and normalize cellular uptake of PFA in linoleic and linolenic LDL via apoВ-100 endocytosis. Atherosclerosis is a syndrome of cellular deficiency of essential polyenic FA.

Keyword: metabolism

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated inflammation.

Keyword: metabolism

Pseudomonas putida Δ9-fatty desaturase: Gene cloning, expression, and function in the cationic surfactants stress.

Pseudomonas putida counteract the fluidizing effect of cationic surfactants decreasing the content of membrane unsaturated fatty (UFA). A Δ9-fatty desaturase gene (desA) from P. putida was isolated, cloned, and successfully expressed in Escherichia coli, a Δ9 desaturase deficient organism. desA consists of 1185\u2009bp and codes for 394 amino acids. The deduced amino sequence reveals three histidine clusters and a hydropathy profile, typical of membrane-bound desaturases. Validating desA expression in E. coli cells, the amount of palmitoleic increased from 2.05 to 7.36%, with the concomitant increase in membrane fluidity (fluorescence polarization value decrease from 0.13\u2009±\u20090.03 to 0.09\u2009±\u20090.02). Also, when DesA activity was assayed in vivo, the percentage of UFA obtained from exogenous [1- C] increased 10-fold. In contrast, when cells expressing desA were exposed 15\u2009min at sublethal concentration of cationic surfactants, the amount of UFA was 82% lower than that detected in cells non-exposed. Thus, the decrease in UFA content to counteract the fluidizing effect of cationic surfactants can be correlated with reduction of DesA activity.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: metabolism

modulation predicts heart failure tests performance.

The changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Pathway Analysis (MetPA) used predictors to identify the most relevant associated to the study, aminoacyl-tRNA and amino biosynthesis, amino , nitrogen , pantothenate and CoA biosynthesis, sphingolipid and glycerolipid , fatty biosynthesis, glutathione , and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the underpinning of CPET and HFBio.

Keyword: metabolism

Effect of co-culture with Tetragenococcus halophilus on the physiological characterization and transcription profiling of Zygosaccharomyces rouxii.

Zygosaccharomyces rouxii and Tetragenococcus halophilus are widely existed and play vital roles during the manufacture of fermented foods such as soy sauce. The aim of this study was to elucidate the effect of T. halophilus CGMCC 3792 on the physiological characterizations and transcription profiling of Z. rouxii CGMCC 3791. Salt tolerance analysis revealed that co-culture with T. halophilus enhanced the salt tolerance of Z. rouxii during salt stress. Analysis of the volatile compounds revealed that co-culture reduced the level of 1-butanol, improved the level of octanoic which all were produced by T. halophilus and reduced the level of phenylethyl alcohol produced by Z. rouxii. The presence of Z. rouxii decreased the contents of 3,4-dimethylbenzaldehyde and acetic produced by T. halophilus. In addition, co-culture improved the content of benzyl alcohol significantly. Analysis of membrane fatty showed that co-culture improved the content of (C16:0) and stearic (C18:0) acids in cells of Z. rouxii, and reduced the contents of myristic (C14:0), palmitoleic (C16:1) and oleic (C18:1). In order to further explore the interactions between the two strains, RNA-seq technology was used to investigate the effect of co-culture with T. halophilus on the transcription profiling of Z. rouxii. By comparing cells incubated in co-culture group with cells incubated in single-culture group, a total of 967 genes were considered as differentially expressed genes (DEGs). Among the DEGs, 72 genes were up-regulated, while 895 genes were down-regulated. These DEGs took party in various activities in cells of Z. rouxii, and the result showed co-culture with T. halophilus had a positive effect on proteolysis, the attachment of a cell to another cell, extracellular protein accumulation, energy , and a negative effect on oxidative phosphorylation, small molecular substances , DNA replication and repair, and transcription in cells of Z. rouxii. Results presented in this study may contribute to further understand the interactions between Zygosaccharomyces rouxii and Tetragenococcus halophilus.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA into herbivorous dry land-living species with oleic type of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: metabolism

Oxidized unsaturated fatty acids induce apoptotic cell death in cultured cells.

Polyunsaturated fatty acids are oxidized by non‑enzymatic or enzymatic reactions. The oxidized products are multifunctional. In this study, we investigated how oxidized fatty acids inhibit cell proliferation in cultured cells. We used polyunsaturated and saturated fatty acids, docosahexaenoic (DHA; 22:6), eicosapentaenoic (EPA; 20:5), linoleic (LA; 18:2), and (16:0). Oxidized fatty acids were produced by autoxidation of fatty acids for 2\xa0days in the presence of a gas mixture (20% O2 and 80% N2). We found that oxidized polyunsaturated fatty acids (OxDHA, OxEPA and OxLA) inhibited cell proliferation much more effectively compared with un‑oxidized fatty acids (DHA, EPA and LA, respectively) in THP‑1 (a human monocytic leukemia cell line) and DLD‑1 (a human colorectal cancer cell line) cells. In particular, OxDHA markedly inhibited cell proliferation. DHA has the largest number of double bonds and is most susceptible to oxidation among the fatty acids. OxDHA has the largest number of highly active oxidized products. Therefore, the oxidative levels of fatty acids are associated with the anti‑proliferative activity. Moreover, caspase‑3/7 was activated in the cells treated with OxDHA, but not in those treated with DHA. A pan‑caspase inhibitor (zVAD‑fmk) reduced the cell death induced by OxDHA. These results indicated that oxidized products from polyunsaturated fatty acids induced apoptosis in cultured cells. Collectively, the switch between cell survival and cell death may be regulated by the activity and/or number of oxidized products from polyunsaturated fatty acids.

Keyword: metabolism

Evidence for perturbed patterns in bipolar disorder subjects associated with lithium responsiveness.

Bipolar disorder (BD) is multifactorial mood disorder characterized by alternating episodes of hyperactive mania and severe depression. Lithium is one of the most preferred drug used as mood stabilizer in treating BD. In this study, we examined the changes in plasma metabolome in BD subjects in the context of lithium responsiveness. Plasma samples from clinically defined, age and gender matched unrelated healthy controls and BD subjects (lithium responders and non-responders) were obtained and processed in positive and negative mode using untargeted liquid chromatography/mass spectrometry analysis. We identified significant alterations in plasma levels of dopamine along with its precursors (tyrosine and phenylalanine), branched chain amino such as valine and excitatory neurotransmitter glutamate between healthy control and BD subjects. Lipid molecules such as, eicosenoic and retinyl ester also showed distinguished patterns between control and BD individuals. Lithium responsiveness was markedly associated with significant differences in proline, L-gamma-glutamyl-isoleucine, dopamine, methyl ester, cholesterol sulfate, androsterone sulfate and 9S,12S,13S-triHOME levels. Altered metabolites enriched with key biochemical associated with neuropsychiatry disorders. We hypothesize that BD pathogenesis and lithium responsiveness is associated with impaired homeostasis of amino and lipid .Copyright © 2019. Published by Elsevier B.V.

Keyword: metabolism

A Peptide-Based HIV-1 Fusion Inhibitor with Two Tail-Anchors and Exhibits Substantially Improved In Vitro and Ex Vivo Anti-HIV-1 Activity and Prolonged In Vivo Half-Life.

Enfuvirtide (T20) is the first U.S. FDA-approved HIV fusion inhibitor-based anti-HIV drug. Its clinical application is limited because of its low potency and short half-life. We previously reported that peptide HP23-E6-IDL, containing both N- and C-terminal anchor-tails, exhibited stronger potency and a better resistance profile than T20. Here we designed an analogous peptide, YIK, by introducing a mutation, T639I, and then a lipopeptide, YIK-C16, by adding (C16) at the C-terminus of YIK. We found that YIK-C16 was 4.4- and 3.6-fold more potent than HP23-E6-IDL and YIK against HIV-1 infection and 13.3- and 10.5-fold more effective than HP23-E6-IDL and YIK against HIV-1 infection, respectively. Consistently, the ex vivo anti-HIV-1 activity, as determined by the highest dilution-fold of the serum causing 50% inhibition of HIV-1 infection, of YIK-C16 in the sera of pretreated mice was remarkably higher than that of YIK or HP23-E6-IDL. The serum half-life (t = 5.9 h) of YIK-C16 was also significantly longer than that of YIK (t = 1.3 h) and HP23-E6-IDL (t = 1.0 h). These results suggest that the lipopeptide YIK-C16 shows promise for further development as a new anti-HIV drug with improved anti-HIV-1 activity and a prolonged half-life.

Keyword: metabolism

The effect of enterolactone on liver lipid precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on lipid fractions fatty acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free fatty acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their fatty acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of fatty acids were evaluated by Western Blot.Enterolactone modified fatty acids composition in FFA, DAG and TAG fractions. In conjunction with lipid overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated fatty acids in each of the investigated lipid fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Rare ginsenosides ameliorate lipid overload-induced myocardial insulin resistance via modulating flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 diabetes. However, their pharmacological studies regarding myocardial fuel and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high fat diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of lipid overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: metabolism

Metabolomics analysis of the protective effect of rubusoside on -induced lipotoxicity in INS-1 cells using UPLC-Q/TOF MS.

Diabetes is one of the most severe chronic diseases worldwide. It is widely accepted that apoptosis of the pancreatic beta cell is an important cause for the induction of hyperglycemia and high levels of free fatty acids (FFAs), also called lipotoxicity associated with pancreatic beta cell dysfunction. Lipotoxicity has been proven to be an important pathogenic factor of diabetes. However, until now, the mechanism of FFA-induced lipotoxicity in INS-1 cells has not been fully understood. Current anti-diabetic drugs that protect islet cells are often toxic to healthy cells, resulting in negative side effects. Thus, there is an urgent need to identify more effective and safer anti-diabetic agents to protect pancreatic islet cells. Rubusoside (RUB) is a major ingredient in the leaves of Rubus suavissimus S. Lee, which decreases blood glucose levels by protecting pancreatic islet cells. However, the exact mechanism of this effect is unknown. In this study, metabolomics experiments based on UPLC-Q/TOF MS characterized a total of 15 metabolites as potential biomarkers associated with lipotoxicity induced by in INS-1 cells. According to the pathway analysis, pentose and glucuronate interconversions, and glycerophospholipid were recognized as the most influenced associated with lipotoxicity. Unexpectedly, deviations of 14 metabolites in lipotoxic INS-1 cells were regulated by RUB, suggesting synergistic mediation of the abnormal . The metabolomics strategy based on UPLC/Q-TOF MS analysis provides a new insight into the mechanisms of lipotoxicity induced by and the anti-lipotoxic activity of RUB in INS-1 cells.

Keyword: metabolism

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: metabolism

Enterobacter sp. strain Fs-11 adapted to diverse ecological conditions and promoted sunflower achene yield, nutrient uptake, and oil contents.

Plant growth-promoting rhizobacteria are under extensive investigation to supplement the chemical fertilizers due to cost-effective and eco-friendly nature. However, their consistency in heterogeneous soil and diverse ecological settings is unclear. The current study presents in vitro and field evaluation of pre-characterized PGPR strain Enterobacter sp. Fs-11 (GenBank accession # GQ179978) in terms of its potential to enhance sunflower yield and oil contents under diverse environmental conditions. Under in vitro conditions, strain Fs-11 showed optimal growth at a range of temperature (15 to 40\xa0°C) and pH values (6.5 to 8.5). Extracellular and intracellular localizations of the strain Fs-11 in sunflower root cortical cells through transmission electron microscopy confirmed its epiphytic and endophytic colonization patterns, respectively. In field experiments, conducted at three different agro-climatic locations, inoculation of strain Fs-11 at 50% reduced NP fertilizer resulted in a significant increase in growth, achene yield, nutrient uptake, and oil contents. Inoculation also responded significantly in terms of increase in mono- and polyunsaturated fatty acids (oleic and linoleic acids, respectively) without rising saturated fatty ( and stearic acids) contents. We concluded that Enterobacter sp. Fs-11 is a potential candidate for biofertilizer formulations to supplement chemical fertilizer requirements of sunflower crop under diverse climatic conditions.

Keyword: metabolism

Permethrin and ivermectin modulate lipid in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid . The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic lipid may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Reporting temporal fluctuations of hepatic C16 and C18 fatty acids during late gestation and early lactation in dromedary camel.

Based on current knowledge, C16 and C18 fatty acids (FA) are considered the most functional FA in hepatic . Although these FAs have been satisfyingly investigated in cattle, other species such as camel have been neglected. For this reason, the current study was designed to scrutinize changing patterns of C16 and C18 FAs in 10 dromedary camels from the last 2\xa0months of gestation to the first\xa0months of lactation. Camels were grazed on natural pasture and supplemented with a balanced ration. Liver biopsies were obtained through blind biopsy technique at about 60, 45, 30, and 15-day antepartum (AP), and at 3, 15, 30, 45, and 60 post-partum (PP). Data were analyzed by the ANOVA procedure of SPSS with repeated measurements. From 15-day AP, saturated FA content of the liver declined (P\u2009<\u20090.01) and 15-day PP reached its peak (P\u2009=\u20090.02). At 30-day PP it went down (P\u2009<\u20090.01), and re-elevated at 45-day PP (P\u2009<\u20090.01) but remained at a steady state for the duration of the study. Mono-unsaturated and polyunsaturated FA content of hepatic tissue were constant throughout AP, albeit observed to peak at 15-day AP compared with 45 (P\u2009=\u20090.04) and 30-day AP (P\u2009<\u20090.01) for mono-unsaturated FAs, and with 60-, 45-, and 30-day AP (P\u2009≤\u20090.01) for polyunsaturated FAs. The content of the liver reached a nadir at 30-day AP (P\u2009<\u20090.01), increased sharply (P\u2009<\u20090.01) at the next sampling time-point, and had a trend to escalate until 3-day PP. Palmitoleic levels were unchanged from 60- to 30-day AP, decreased at 15 AP and 3-day PP, increased at 15-day PP, then remained constant until the end of the study period (P\u2009≤\u20090.04). Stearic content started to grow at 15-day AP and reached its peak at 15-day PP (P\u2009<\u20090.01). At 30-day PP, stearic level in liver dropped abruptly (P\u2009<\u20090.01), then intensified at 45-day PP and did not change after; hepatic content of stearic was lower during AP compared with PP time-points. Other C18 FAs changed significantly during the study period. These results suggest that parturition could have a profound effect on FA composition and other metabolites in camel liver. Further research is required to establish the mechanism behind these changes.

Keyword: metabolism

Replication of Marek\'s Disease Virus Is Dependent on Synthesis of Fatty and Prostaglandin E.

Marek\'s disease virus (MDV) causes deadly lymphoma and induces an imbalance of the lipid in infected chickens. Here, we discovered that MDV activates the fatty synthesis (FAS) pathway in primary chicken embryo fibroblasts (CEFs). In addition, MDV-infected cells contained high levels of fatty acids and showed increased numbers of lipid droplets (LDs). Chemical inhibitors of the FAS pathway (TOFA and C75) reduced MDV titers by approximately 30-fold. Addition of the downstream metabolites, including malonyl-coenzyme A and , completely restored the inhibitory effects of the FAS inhibitors. Furthermore, we could demonstrate that MDV infection activates the COX-2/prostaglandin E (PGE) pathway, as evident by increased levels of arachidonic , COX-2 expression, and PGE synthesis. Inhibition of the COX-2/PGE pathway by chemical inhibitors or knockdown of COX2 using short hairpin RNA reduced MDV titers, suggesting that COX-2 promotes virus replication. Exogenous PGE completely restored the inhibition of the COX-2/PGE pathway in MDV replication. Unexpectedly, exogenous PGE also partially rescued the inhibitory effects of FAS inhibitors on MDV replication, suggesting that there is a link between these two in MDV infection. Taken together, our data demonstrate that the FAS and COX-2/PGE play an important role in the replication of this deadly pathogen. Disturbances of the lipid in chickens infected with MDV contribute to the pathogenesis of disease. However, the role of lipid in MDV replication remained unknown. Here, we demonstrate that MDV infection activates FAS and induces LD formation. Moreover, our results demonstrate that MDV replication is highly dependent on the FAS pathway and the downstream metabolites. Finally, our results reveal that MDV also activates the COX-2/PGE pathway, which supports MDV replication by activating PGE/EP2 and PGE/EP4 signaling .Copyright © 2019 Boodhoo et al.

Keyword: metabolism

miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: metabolism

Novel hypolipidemic conjugates of fatty and bile with lysine for linkage.

Novel fatty -bile conjugates (1a-1k) were designed and synthesized by coupling of the fatty acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve liver-specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high-fat diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced liver damage.

Keyword: metabolism

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced Lipogenesis by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells.

4EBP1 is a chief downstream factor of mTORC1, and PPARγ is a key lipogenesis-related transcription factor. mTORC1 and PPARγ are associated with lipid . However, it is unknown which effector protein connects mTORC1 and PPARγ. This study investigated the interaction between 4EBP1 with PPARγ as part of the underlying mechanism by which insulin-induced lipid synthesis and secretion are regulated by mTORC1 in primary bovine mammary epithelial cells (pBMECs). Rapamycin, a specific inhibitor of mTORC1, downregulated 4EBP1 phosphorylation and the expression of PPARγ and the following lipogenic genes: lipin 1, DGAT1, ACC, and FAS. Rapamycin also decreased the levels of intracellular triacylglycerol (TAG); 10 types of fatty ; and the accumulation of TAG, (PA), and stearic (SA) in the cell culture medium. Inactivation of mTORC1 by shRaptor or shRheb attenuated the synthesis and secretion of TAG and PA. In contrast, activation of mTORC1 by Rheb overexpression promoted 4EBP1 phosphorylation and PPARγ expression and upregulated the mRNA and protein levels of lipin 1, DGAT1, ACC, and FAS, whereas the levels of intracellular and extracellular TAG, PA, and SA also rose. Further, 4EBP1 interacted directly with PPARγ. Inactivation of mTORC1 by shRaptor prevented the nuclear location of PPARγ. These results demonstrate that mTORC1 regulates lipid synthesis and secretion by inducing the expression of lipin 1, DGAT1, ACC, and FAS, which is likely mediated by the 4EBP1/PPARγ axis. This finding constitutes a novel mechanism by which lipid synthesis and secretion are regulated in pBMECs.

Keyword: metabolism

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 .

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic . DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: metabolism

Valorization of crude glycerol based on biological processes for accumulation of lipophilic compounds.

Bacteria that are capable of accumulating lipids in their cells as storage compounds can also produce polyhydroxyalkanoates of high technological value, depending on the specific culture conditions. The objective of this study was to utilize crude glycerol from biodiesel (CGB) as a substrate, which is a major byproduct from biodiesel production, to produce lipophilic compounds. Bacillus megaterium INCQS 425 was cultivated and evaluated for the production of lipophilic compounds and the properties of these compounds were investigated. Cultivation of the bacteria in a medium with a C:N ratio of 0.60:1 favored the accumulation of lipids by (17.5%) comprising mainly (13.08%), palmitoleic (39.48%), and especially oleic (37.02%), which imparts good characteristics to biodiesel. Meanwhile, cultivation of the bacteria in a medium with a C:N ratio of 4:1 favored the accumulation of polyhydroxyalkanoates (PHA) (3.31gL) mainly comprising medium and long chain PHA. Low crystallinity (<30%) and excellent thermal properties make them suitable for processes that demand high temperatures, such as extrusion. The lipids produced in the present study had satisfactory oxidative stability for the production of quality biodiesel. The polyhydroxyalkanoates produced in the study are of low cost and have promising thermal properties that justify its technological potential, thereby configuring highly competitive bioproducts.Copyright © 2019. Published by Elsevier B.V.

Keyword: metabolism

Modeling growth and fermentation inhibition during bioethanol production using component profiles obtained by performing comprehensive targeted and non-targeted analyses.

Corn cob and corn stover hydrolysates are forms of lignocellulosic biomass that can be used in second generation bioethanol production and biorefinery processes. Growth and fermentation inhibitors generated during physicochemical and enzymatic hydrolysis decrease ethanol and biomaterial production during the subsequent biological processes. Here, estimates of growth and fermentation inhibition during bioethanol fermentation were made using component profiles of corn cobs and corn stover at different degrees of hydrolysis. The component profiles were acquired by non-targeted gas chromatography mass spectrometry and targeted high-performance liquid chromatography. Correlations between the comprehensive analysis results and yeast growth and ethanol production were modeled very accurately by partial-least-squares regression analysis. Acetate, apocynin, butyrovanillone, furfural, furyl hydroxymethyl ketone, m-methoxyacetophenone, , syringaldehyde, and xylose, were compounds with very variable importance in projection values and had negative correlation coefficients in the model. In fact, methoxyacetophenone, apocynin, and syringaldehyde inhibited fermentation more than furfural in equivalent concentration.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Cyclophilin D participates in the inhibitory effect of high-fat diet on the expression of steroidogenic acute regulatory protein.

The high-fat diet (HFD)-induced obesity is responsible for the testosterone deficiency (TD). However, the mechanism remains unknown. Mitochondrial homeostasis is proved to be important for maintaining the function of steroidogenic acute regulatory protein (StAR), the first rate-limiting enzyme in testosterone synthesis. As the key regulator of mitochondrial membrane permeability, cyclophilin D (CypD) plays a crucial role in maintaining mitochondrial function. In this study, we sought to elucidate the role of CypD in the expression of StAR affected by HFD.To analyse the influence of CypD on StAR in vivo and in vitro, mouse models of HFD, CypD overexpression and CypD knockout (Ppif ) as well as Leydig cells treated with (PA) and CypD overexpression plasmids were examined with an array of , mitochondrial function and molecular assays.Compared with the normal diet mice, consistent with reduced testosterone in testes, the expressions of StAR in both mRNA and protein levels in HFD mice were down-regulated, while expressions of CypD were up-regulated. High-fat intake impaired mitochondrial function with the decrease in StAR in Leydig cells. Overexpression of CypD inhibited StAR expressions in vivo and in vitro. Compared with C57BL/6 mice with HFD, expressions of StAR were improved in Ppif mice with HFD.Mitochondrial CypD involved in the inhibitory effect of HFD on StAR expression in testes.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: metabolism

Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether obesity impacts placental lipid transport and remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and insulin-sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal obesity. Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in lipid classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to insulin resistance and low-grade inflammation in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by obesity.-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

Keyword: metabolism

Deciphering the intervention mechanism of Taohong Siwu Decoction following the abnormal uterine bleeding rats based on serum profiles.

Abnormal uterine bleeding (AUB), one of the most significant characters of incomplete abortion, is a widespread phenomenon in gynecological that put a woman into a terrible physiological and psychological state. Taohong Siwu Decoction (TSD) is a traditional Chinese medicine (TCM) prescriptions which have treated AUB in China for decades. Our previous study elucidated that TSD reduced the volume of uterine bleedings as well as repaired the endometrium. The present study aims to investigate the mechanisms of TSD on AUB based on serum metabolomics. In this study, serum profile data was collected using ultra high-performance liquid chromatography with ion trap/time-of-flight mass spectrometry and gas chromatography-mass spectrometry. 23 potential biomarkers (urea, serine, L-proline, L-glutamic , , l-acetylcarnitine, LysoPC(16:0), LysoPC(20:4), l-proline, linoleic , stearic , l-isoleucine, phenylalanine, l-tyrosine, Oleic , et al) were eventually identified using multivariate statistical analysis (PCA and OPLS-DA) with VIP\u2009>\u20091, P\u2009<\u20090.05. Correlation analysis, fold-change (FC), area under receiver characteristic (ROC), false discovery rate (FDR) were used for data confirmation to ensure the authenticity of the data. The related- pathway mainly included amino (Phenylalanine, tyrosine, and tryptophan ; Valine, leucine and isoleucine biosynthesis; Arginine and proline ; Glycine, serine and threonine ) and lipid (linoleic , glycerophospholipid ). The results show that TSD has a favorable therapeutic effect on AUB by adjusting the disorders, which could provide dietary guidance for the clinic.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolism

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic fatty liver disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling .

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid , liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal .Copyright © 2019. Published by Elsevier B.V.

Keyword: metabolism

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host energy through the regulation of body fat storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: metabolism

[Psoralen and isopsoralen improve lipid disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: metabolism

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid -related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of lipid - and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid -related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid , suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Physicochemical characteristics of fat blend from hydrogenated coconut oil and acyl migrated palm mid-fraction.

Palm mid-fraction (PMF), which has a high content of symmetric POP, was converted to asymmetric PPO (APMF) via acyl migration. After solvent fractionation, the liquid phase of acyl migrated PMF (APMF-L) was obtained and blended with hydrogenated coconut oil (HCO, 50:50, w/w) to produce a fat blend (namely, an alternative fat blend) which had reduced saturated fatty content while having similar melting behavior to HCO. In an alternative fat blend, the major fatty acids were lauric (27.94), (26.93) and oleic (15.75\u202fmol%) . The solid fat index was quite similar to that of HCO, especially at 28-44\u202f°C. Nevertheless, an alternative fat blend had lower saturated fatty content, by 18%, compared to HCO. The content of highly atherogenic myristic was reduced by approximately 40%. The alternative fat blend in this study could be used as a raw material for non-dairy cream with low saturated fat content.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Cypate and Cypate-Glucosamine as Near-Infrared Fluorescent Probes for In Vivo Tumor Imaging.

Near-infrared (NIR) imaging is a promising technique for use as a noninvasive and sensitive diagnostic tool. Although the NIR fluorescently labeled glucose analog glucosamine (cypate-glucosamine) has applications in preclinical imaging, the transport and fate of this probe in tissues remain unaddressed. Here, we have synthesized and characterized cypate and cypate-glucosamine conjugate (cy-2-glu), and investigated the probable transport of these probes in vitro and in vivo. We compared uptake of the probes in the presence and absence of excess d-glucose, "saturated cypate" and in two normal-cancer cell line pairs: lung cancer (A549)-normal (MRC9) and prostate cancer (DU145)-normal (BPH). Breast cancer (MDA-MB-231) and liver cancer (HepG2) cell lines were also examined. Results support use of the glucose transport pathway by cy-2-glu and fatty transport pathway by cypate. Mass spectrometry data on the in vitro extracts revealed deamidation of cy-2-glu in prostate and liver cells, suggesting release of glucosamine. In vivo biodistribution studies in mice engrafted with breast tumors showed a distinct accumulation of cy-2-glu in liver and tumors, and to a lesser extent in kidneys and spleen. A negligible accumulation of cypate alone in tumors was observed. Analysis of urine extracts revealed renal excretion of the cy-2-glu probe in the form of free cypate, indicating deamidation of cy-2-glu in tissues. Thus, investigation of the used by NIR probes such as cy-2-glu advances their use in the detection and monitoring of tumor progression in preclinical animal studies.Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: metabolism

A high-fat diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high-fat diet (HFD) commonly leads to obesity and insulin resistance. However, even 3\u2009days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus.Mice were fed either a low-fat diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from fat for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: metabolism

Prepartum fatty supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb liver and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb liver and adipose tissue fatty (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and liver and adipose tissue collected for FA and gene expression analysis. Liver concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in liver tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), fatty synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in liver of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain fatty 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that lipogenesis decreased, and lipolysis increased in lamb liver with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that lipogenesis decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not liver, to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: metabolism

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces , oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on -, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and .

Keyword: metabolism

Adverse Effects in Skeletal Muscle Following the Medicinal Use of Nicotiana glauca.

Nicotiana glauca is a cosmopolitan shrub, used in medicine to treat swellings, wounds, sores and cancer. However, its users lack of knowledge of the adverse effects. We seek to evaluate the effects of lipid extracts from N. glauca on myoblasts, identifying the compounds which cause undesirable effects. Myoblasts are important in muscle homeostasis, thus a high death rate of them cause myopathies. We performed an ethanolic extraction from leaves of N. glauca and the extract was successively partitioned with hexane, chloroform and ethyl acetate. The effects of extracts in C2C12 cells were analysed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Mitotracker and 4\',6-diamidino-2-phenylindole (DAPI) staining, Western blotting, real-time PCR and immunofluorescence assays. Caspase activity was studied. The fraction with the highest apoptotic effects was analysed by chromatography, NMR and GC-MS spectrometry were used to identify the apoptotic agent, after which its biological activity was evaluated. The extracts from N. glauca induced apoptosis in C2C12 cells involving caspase-3/7. We found that the extracts trigger a defence response in muscle through Akt and heat shock protein 27 (HSP27). We identified an apoptotic agent as . These data suggest that the use of N. glauca in hormone replacement therapy, or in other therapies affects skeletal muscle homeostasis, worsening the negative effects of the menopause. Thus, the relevance of this work lies in the fact that it is the first time that a report about the molecular mechanism responsible for the side effects of medicinal use of N. glauca, has been shown. Moreover the compound responsible for these effects has been identified.

Keyword: metabolism

Human cytochrome P450 enzymes bind drugs and other substrates mainly through conformational-selection modes.

Cytochrome P450 (P450) enzymes are major catalysts involved in the oxidations of most drugs, steroids, carcinogens, fat-soluble vitamins, and natural products. The binding of substrates to some of the 57 human P450s and other mammalian P450s is more complex than a two-state system and has been proposed to involve mechanisms such as multiple ligand occupancy, induced-fit, and conformational-selection. Here, we used kinetic analysis of binding with multiple concentrations of substrates and computational modeling of these data to discern possible binding modes of several human P450s. We observed that P450 2D6 binds its ligand rolapitant in a mechanism involving conformational-selection. P450 4A11 bound the substrate lauric via conformational-selection, as did P450 2C8 with . Binding of the steroid progesterone to P450 21A2 was also best described by a conformational-selection model. Hexyl isonicotinate binding to P450 2E1 could be described by either a conformational-selection or an induced-fit model. Simulation of the binding of the ligands midazolam, bromocriptine, testosterone, and ketoconazole to P450 3A4 was consistent with an induced-fit or a conformational-selection model, but the concentration dependence of binding rates for varying both P450 3A4 and midazolam concentrations revealed discordance in the parameters, indicative of conformational-selection. Binding of the P450s 2C8, 2D6, 3A4, 4A11, and 21A2 was best described by conformational-selection, and P450 2E1 appeared to fit either mode. These findings highlight the complexity of human P450-substrate interactions and that conformational-selection is a dominant feature of many of these interactions.© 2019 Guengerich et al.

Keyword: metabolism

Mechanisms of electrical remodeling in lipotoxic guinea pig heart.

To develop an adult guinea pig model of lipotoxicity and explore the underlying mechanisms associated with changes in the expression of the delayed rectifier potassium current (I).Lipotoxicity may represent a common link among disorders and a higher vulnerability to arrhythmias.Whole-cell patch clamp, and (PA, a potent inducer of lipotoxicity), were used to assess mechanisms of short-term (∼50 days) high-fat diet (HFD) feeding on atrial electrophysiology in guinea pig hearts and myocytes.HFD fed guinea pigs were significantly heavier, displayed hypertriglyceridemia and hypercholesterolemia; but no signs of hyperglycemia or inflammation compared to low-fat diet fed controls. Increasing cardiac PA levels, resulted in shortened atrial action potential duration, and increased I density. Inhibition of phosphoinositide 3-kinase (PI3K) prevented increases in I due to PA. Acute (≥1hr) exposure of atrial myocytes to exogenous PA (1\u202fmM) increased the density of the rapid delayed rectifier potassium current I, while it was decreased with the unsaturated oleic (OA, 1\u202fmM). Serine-threonine protein phosphatase-2 (PP2A) inhibition with cantharidin reversed the effect of OA on I.Our data provide evidence of a novel lipotoxic guinea pig model with signs of vulnerability to arrhythmias. Inhibition of PA/PI3K/I and/or activation of the OA/PP2A/I may be therapeutically beneficial for lipotoxic arrhythmias.Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

Inhibition of soluble epoxide hydrolase attenuates a high-fat diet-mediated renal injury by activating PAX2 and AMPK.

A high-fat diet (HFD) causes obesity-associated morbidities involved in macroautophagy and chaperone-mediated autophagy (CMA). AMPK, the mediator of macroautophage, has been reported to be inactivated in HFD-caused renal injury. However, PAX2, the mediator for CMA, has not been reported in HFD-caused renal injury. Here we report that HFD-caused renal injury involved the inactivation of Pax2 and Ampk, and the activation of soluble epoxide hydrolase (sEH), in a murine model. Specifically, mice fed on an HFD for 2, 4, and 8 wk showed time-dependent renal injury, the significant decrease in renal Pax2 and Ampk at both mRNA and protein levels, and a significant increase in renal sEH at mRNA, protein, and molecular levels. Also, administration of an sEH inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea, significantly attenuated the HFD-caused renal injury, decreased renal sEH consistently at mRNA and protein levels, modified the renal levels of sEH-mediated epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) as expected, and increased renal Pax2 and Ampk at mRNA and/or protein levels. Furthermore, (PA) treatment caused significant increase in , and decrease in both Pax2 and Ampk in murine renal mesangial cells (mRMCs) time- and dose-dependently. Also, 14(15)-EET (a major substrate of sEH), but not its sEH-mediated metabolite 14,15-DHET, significantly reversed PA-induced increase in , and PA-induced decrease in Pax2 and Ampk. In addition, plasmid construction revealed that Pax2 may positively regulate Ampk transcriptionally in mRMCs. This study provides insights into and therapeutic target for the HFD-mediated renal injury.

Keyword: metabolism

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: metabolism

Outdoor cultivation of the green microalga Chlorella vulgaris under stress conditions as a feedstock for biofuel.

The present work investigated the potential of the green alga Chlorella vulgaris to produce high-quality biofuel under culture stress conditions. The cultivation was carried out in a 1000 l\xa0open plate tank system, which provides biomass yields comparable to open pond systems, but with less area needed. Algal biomass and lipid content were measured repeatedly. We compared the two solvent systems n-hexane and hexane/isopropanol (HIP) for extraction efficiency of lipids and applied three different extraction methods Soxhlet, soaking, and soaking followed by Soxhlet (soak-Sox). The combination of the HIP solvent and the soak-Sox provided the highest lipid yield (15.8\u2009±\u20090.174). Volumetric biomass and lipid productivity were 0.201\xa0g\xa0l\xa0day and 31.71\xa0mg\xa0l\xa0day, respectively, whereas areal biomass and lipid productivity were 25.73\xa0g\xa0m\xa0day and 4.066\xa0g\xa0m\xa0day, respectively. The fatty profile by means of gas chromatography resulted in seven fatty acids from C to C. The most abundant fatty methyl esters (FAMES) were (C16:0), oleic (C18:1), and stearic (C18:0) acids. Lipid synthesis enhanced by optimizing the Kuhl growth medium with replacing nitrate by urea (50% N compared to the original recipe) increased salt content (10\xa0g/l NaCl), ferrous sulfate (0.5\xa0g/l), and sodium acetate addition (1\xa0g/l). With regard to density, kinematic viscosity, gravity, pour point, flash point, and cetane number, the Chlorella-biodiesel comply with ASTM and EN standards thus pointing at the high potential of lipids synthesized by Chlorella as a feedstock for biodiesel production.

Keyword: metabolism

Berberine attenuates nonalcoholic hepatic steatosis through the AMPK-SREBP-1c-SCD1 pathway.

Berberine (BBR), a natural compound extracted from Chinese herb, has been shown to effectively attenuate nonalcoholic fatty liver disease (NAFLD) in clinic. However, the mechanism underlying the effect of BBR is not fully understood. Stearyl-coenzyme A desaturase 1 (SCD1) mediates lipid in liver. Therefore, we hypothesized that SCD1 mediated the beneficial effect of BBR on NAFLD.The expression of SCD1 was measured in the liver of NAFLD patients and ob/ob mice. The effect of BBR on NAFLD was evaluated in C57BL/6\u202fJ mice on high fat diet (HFD). The effect of BBR was also investigated in HepG2 and AML12\u202fcells exposed to high glucose and . Oil red O staining was performed to detect triglyceride (TG) level. Quantitative real-time polymerase chain reaction and Western blot were used to detect the messenger ribonucleic (mRNA) and protein expression of target genes. The activity of SCD1 promoter was measured by dual-luciferase reporter assay.The expression of SCD1 was increased in the liver of NAFLD patients and ob/ob mice. BBR reduced hepatic TG accumulation and decreased the expressions of hepatic SCD1 and other TG synthesis related genes both in vivo and in vitro. Knockdown of SCD1 expression mimicked the effect of BBR decreasing TG level in steatotic hepatocytes, whereas overexpression of SCD1 attenuated the effect of BBR. Mechanistically, BBR promoted the phosphorylation of AMP-activated protein kinase (AMPK) and sterol regulatory element-binding protein-1c (SREBP-1c) in HepG2 cells and the liver of HFD-fed mice. Activation of the AMPK-SREBP-1c pathway and sterol regulatory element (SRE) motif in SCD1 promoter (-920/-550) was responsible for the BBR-induced suppression of SCD1.BBR reduces liver TG synthesis and attenuates hepatic steatosis through the activation of AMPK-SREBP-1c-SCD1 pathway.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Inhibitory effect of 17β‑estradiol on triglyceride synthesis in skeletal muscle cells is dependent on ESR1 and not ESR2.

The present study aimed to investigate the inhibitory effects and the mechanisms underlying 17β‑estradiol (E2) effects on triglyceride synthesis and insulin resistance in skeletal muscle tissues and cells. Ovariectomy (OVX) was performed on 6‑month‑old female rats treated with or without E2. Subsequently, various serum biochemical markers were measured. Additionally, pathological alterations of the uterus, liver and skeletal muscle were analyzed, and the content of triglycerides (TG) in muscle was detected. Differentiated myotubes formed by C2C12 cells were treated with (PA) or pretreated with E2, estrogen receptor (ESR)\xa01 agonist propylpyrazoletriol (PPT) and ESR2 agonist diarylpropionitrile (DPN). Subsequently, the mRNA or protein expression levels of ESR1/2, peroxisome proliferator activated receptor α (PPARα), CD36 molecule (CD36), fatty synthase (FASN), perilipin 2 (PLIN2), phosphorylated acetyl‑CoA carboxylase α (p‑ACACA), p‑AKT serine/threonine kinase (p‑AKT) and p‑mitogen‑activated protein kinase\xa08 (p‑MAPK8) were analyzed in skeletal muscle or in C2C12 cells by reverse transcription‑semi‑quantitative polymerase chain reaction and western blotting. The present results suggested that treatment with E2 inhibited OVX‑induced body weight gain, TG accumulation and insulin resistance. The protein or mRNA expression levels of ESR1, CD36, PPARα, p‑ACACA and p‑AKT were decreased, whereas the protein or mRNA expression levels of ESR2, PLIN2, FASN and p‑MAPK8 were increased in the OVX group. Of note, treatment with E2 restored the expression levels of the aforementioned factors. In C2C12 cells, treatment with E2 or PPT reversed the alterations induced by treatment with PA. In contrast, pretreatment with DPN did not influence the effect of PA. Collectively, E2 was able to interact with ESR1, thus activating the CD36‑PPARα pathway, decreasing the level of TG in the muscles and improving insulin resistance in skeletal muscles and C2C12 cells.

Keyword: metabolism

Effect of clones, year of harvest and geographical origin of fruits on quality and chemical composition of Argan oil.

Argan oil is precious oil with food and cosmetic uses. In recent years, this oil has been subject to an increasing national and international demand. The present work aims at studying the effect of clones and age, year of harvest and geographical origin on Argan oil quality and chemical composition. The results indicate that age does not affect Argan oil quality and fatty acids content. However, clones had a significant effect on fatty acids and tocopherol levels. Saturated and unsaturated fatty acids were highly influenced by the year of harvest and geographical origin, presumably due to climatic conditions. Unsaturated fatty acids varied from 78.28% to 81.77%. Depending on clones, total tocopherols varied from 687.40\u202fmg/kg to 1068\u202fmg/kg. This study is useful for the choice of clones with the aim of developing Argan trees orchards destined to oil production.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high fat diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related diseases such as type II diabetes mellitus (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: metabolism

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/lipid in type 2 diabetes mellitus. Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high-fat diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

Keyword: metabolism

Gut Microbiota and Metabolome Response of Seed Oil on Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut microbiota dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan .

Keyword: metabolism

mTORC2 Regulates Lipogenic Gene Expression through PPAR to Control Lipid Synthesis in Bovine Mammary Epithelial Cells.

The mechanistic target of rapamycin complex 2 (mTORC2) primarily functions as an effector of insulin/PI3K signaling to regulate cell proliferation and is associated with cell . However, the function of mTORC2 in lipid is not well understood. In the present study, mTORC2 was inactivated by the ATP-competitive mTOR inhibitor AZD8055 or shRNA targeting in primary bovine mammary epithelial cells (pBMECs). MTT assay was performed to examine the effect of AZD8055 on cell proliferation. ELISA assay and GC-MS analysis were used to determine the content of lipid. The mRNA and protein expression levels were investigated by RT/real-time PCR and western blot analysis, respectively. We found that cell proliferation, mTORC2 activation, and lipid secretion were inhibited by AZD8055. was knocked down and mTORC2 activation was specifically attenuated by the shRNA. Compared to control cells, the expression of the transcription factor gene and the lipogenic genes , , , and was downregulated in silencing cells. As a result, the content of intracellular triacylglycerol (TAG), (PA), docosahexaenoic (DHA), and other 16 types of fatty was decreased in the treated cells; the accumulation of TAG, PA, and DHA in cell culture medium was also reduced. Overall, mTORC2 plays a critical role in regulating lipogenic gene expression, lipid synthesis, and secretion in pBMECs, and this process probably is through PPAR. This finding provides a model by which lipogenesis is regulated in pBMECs.

Keyword: metabolism

Pre-fermentative supplementation of fatty acids alters the activity of wine yeasts.

Fatty acids play important roles in the maintenance of cell membrane, viability and overall of wine yeasts (particularly Saccharomyces cerevisiae) during adverse winemaking conditions. We previously showed that linoleic supplementation markedly affect aroma compound production of S. cerevisiae wine strains. However, very little is known about how other commonly found fatty acids in grape juice modulate the growth and of S. cerevisiae. We aimed to determine the individual effect of five fatty acids on fermentation patterns and of two wine yeast strains (S. cerevisiae EC1118 and X5). Microvinification was performed at 15\u202f°C by supplementing a grape juice (individually) with three different concentrations of saturated (), unsaturated (oleic, linoleic and γ-linolenic acids) and short-chain (hexanoic ) fatty acids. Metabolite profiles of the resulting wines were determined using Gas-chromatography coupled to Mass-spectrometry (GC-MS). Our data show that the addition of γ-linolenic to the juice caused the production of higher amounts of amino and organic acids (except isoleucine and 2-oxoglutaric ) in wines when fermented by EC1118, while supplementation showed similar trends when fermented by X5. The effect of linoleic was independent of yeast strains and we observed a global reduction of amino and organic acids (except pyruvic ) while increased production of most of the fatty acids other than the supplemented ones. Our data clearly suggest that pre-fermentative supplementation of different fatty acids indeed influenced the growth and of wine yeasts in a different way. Thus, attention needs to be paid not only to the wine yeast strain used during the winemaking but also to the overall grape juice composition, including fatty acids, to obtain the desired wine characteristics.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Dietary modulation of energy homoeostasis and -inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate -inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on -inflammation, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate -inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy is a key determinant of inflammation. Whilst -inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects -inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and processes.

Keyword: metabolism

[Effect and mechanism of liraglutide on the apoptosis of human hepatocellular carcinoma HepG2 cells induced with ].

To observe whether liraglutide protects HepG2 cells from lipotoxicity by affecting mitogen-activated protein kinase (MAPKs) pathway. HepG2 cells were induced with 400μmol/L , and cells were treated with a final concentration of 100 nmol/L liraglutide. In addition, JNK inhibitor (SP600125) and p38 MAPK inhibitor (SB203580) were added in advance, respectively. Apoptosis rate, malondialdehyde (MDA) content, and caspase3 activity were detected. Western blot was used to detect p38 mitogen-activated protein kinase (p38 MAPK), c-jun amino terminal kinase (JNK), cytochrome oxidase P450 2E1 (CYP2E1), glucose regulatory protein 78 (GRP78), activated caspase 3, B cell lymphoma associated Protein X (Bax), B cell lymphoma 2 (Bcl-2), and expression of C/EBP homologous protein (CHOP) protein. LSD or Dunnett\'s T3 test were used to compare the mean of multiple samples. increased the phosphorylation of p38 MAPK and JNK in HepG2 cells (< 0.05). Furthermore, it increased the expression of GRP78, CHOP, CYP2E1, MDA, Bax, caspase3 and apoptosis rate, but inhibited the expression of Bcl-2 (value < 0.05). SP600125 and SB203580 had inhibited oxidative stress and apoptosis induced by (including CYP2E1, MDA, Bax, Bcl-2, caspase3, CHOP) (< 0.05). The phosphorylation level of p38 MAPK and JNK was reduced with liraglutide and the expression of apoptosis-related proteins (Bax, Bcl-2, caspase3, CHOP) (< 0.05) was regulated. There was no significant difference in the effect of liraglutide on apoptotic proteins (Bax, Bcl-2, caspase-3, CHOP) (> 0.05) after pretreatment with those two inhibitors. has strong lipotoxicity to HepG2 cells and induces apoptosis. Glucagon-like peptide-1 analogue, liraglutide may improve lipotoxicity of by mediating p38 MAPK and JNK .

Keyword: metabolism

Influence of nutrient formulations on growth, lipid yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.

The study was conducted to analyse the influence of three nutrient formulations, namely BG-11 medium, BBM and TAP medium, on growth potential and lipid yield of two microalgal genera (Botryococcus sp. and Chlorella sp.) and to study the roles of N, P and other major nutrients. The study focussed on the general patterns of starch and lipid synthesis and storage and to further assess how photosynthetic carbon partitioning into starch and lipid is altered by conditions in growth media such as N and C presence as seen in BG11 medium which are known to induce neutral lipid production and the lack of it in BBM and TAP medium. BG-11 medium performed better as compared to BBM and TAP medium in terms of biomass productivity and lipid yield. The lipid yield was highest in Botryococcus sp. (63.03% dry wt.) and Chlorella sp. (50.27% dry wt.) at 30th day of incubation. Mean biomass productivity was highest for Botryococcus in BBM medium (6.14\xa0mg/L/day) and for Chlorella in BG-11 medium (4.97\xa0mg/L/day). Mean lipid productivity (50.78% and 39.36%) was highest in BG11 medium for both Botryococcus and Chlorella species, respectively. A sharp decline in sugar content was observed in the late stationary phase of growth from 30th day to 45th day. Fatty methyl ester (FAME) profile of the extracted lipids showed predominantly oleic , followed by and stearic in both the strains when grown in BG-11 medium. The other biodiesel quality parameters were in accordance with the international standards. A complex relationship was found between chemical composition and biodiesel properties. Proximity analysis indicated that the fuel properties of biodiesels are determined by a number of parameters and by the combination of different chemical compositions. The results provide an insight into organic carbon partitioning into lipid compounds and how the organism\'s lipid changes due to N-deplete culturing in TAP medium and inorganic carbon source availability as seen in BG-11 and BBM medium.

Keyword: metabolism

Obesity Mediates Apoptosis and Extracellular Matrix Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration.

Obesity may promote intervertebral disc degeneration (IDD) by non-mechanical means, by influencing levels of free fatty acids which could impair cell . This study aims to establish factors in obesity-related IDD independent of mechanical loading. In clinical study, we retrospectively reviewed 128 volunteers (73 males, 55 females, aged 29-88 years) and compared their grades of disk degeneration with obesity-related factors such as body weight, BMI, and serum lipid levels. Clinically, the IDD group showed increased age, BMI and serum triglyceride. Triglyceride was a significant risk factor for IDD even after correction for BMI and age (P = 0.007). In obesity animal model, rats were fed a high-fat diet (HFD) in order to study its effects on disk and apoptosis. HFD rats had significantly higher serum levels of lipids, including triglyceride and non-esterified fatty , and showed significantly decreased markers of anabolism, increased catabolism and apoptosis in disk. Finally, rat nucleus pulposus (NP) cells were stimulated with a fatty (, PA) to gauge its effects on cell and apoptosis. Cell culture studies showed that NP cells exposed to PA showed increased apoptosis for activation of caspase 3, 7, 9, and PARP, which was primarily via the MAPK signal pathway, especially ERK pathway. In conclusion, hypertriglyceridemia can lead to IDD, independently of age and BMI. Hypertriglyceridemia appears to mediate disk cell apoptosis and matrix catabolism primarily via the ERK pathway.Copyright © 2019 Zhang, Chen, Huang, Wang, Shan, Liu, Chen, Li, Fan and Zhao.

Keyword: metabolism

Amelioration of Endoplasmic Reticulum Stress by Mesenchymal Stem Cells via Hepatocyte Growth Factor/c-Met Signaling in Obesity-Associated Kidney Injury.

Recent advances in the understanding of lipid suggest a critical role of endoplasmic reticulum (ER) stress in obesity-induced kidney injury. Hepatocyte growth factor (HGF) is a pleiotropic cytokine frequently featured in stem cell therapy with distinct renotropic benefits. This study aims to define the potential link between human induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs)/bone marrow-derived MSCs (BM-MSCs) and ER stress in lipotoxic kidney injury induced by (PA) in renal tubular cells and by high-fat diet (HFD) in mice. iPS-MSCs or BM-MSCs alleviated ER stress (by preventing induction of Bip, chop, and unfolded protein response), inflammation (Il6, Cxcl1, and Cxcl2), and apoptosis (Bax/Bcl2 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells) in renal cortex of animals exposed to HFD thus mitigating histologic damage and albuminuria, via activating HGF/c-Met paracrine signaling that resulted in enhanced HGF secretion in the glomerular compartment and c-Met expression in the tubules. Coculture experiments identified glomerular endothelial cells (GECs) to be the exclusive source of glomerular HGF when incubated with either iPS-MSCs or BM-MSCs in the presence of PA. Furthermore, both GEC-derived HGF and exogenous recombinant HGF attenuated PA-induced ER stress in cultured tubular cells, and this effect was abrogated by a neutralizing anti-HGF antibody. Taken together, this study is the first to demonstrate that MSCs ameliorate lipotoxic kidney injury via a novel microenvironment-dependent paracrine HGF/c-Met signaling mechanism to suppress ER stress and its downstream pro-inflammatory and pro-apoptotic consequences. Stem Cells Translational Medicine 2019;8:898&910.© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

Keyword: metabolism

Tissue-Specific Distribution of Sphingomyelin Species in Pork Chop Revealed by Matrix-Assisted Laser Desorption/Ionization-Imaging Mass Spectrometry.

Sphingomyelin (SM) species are major sphingolipids in pork meat that affect quality parameters, such as health benefits due to their protective properties against chronic diseases; however, their spatial distribution remains unclear. We used matrix-assisted laser desorption/ionization (MALDI)-imaging mass spectrometry (IMS) to investigate the distribution and composition of SM species in pork chop consisting of longissimus thoracis et lumborum muscle (loin), intermuscular fat tissue, transparent tissue, and spinalis muscle. Four SM species were identified by liquid chromatography-electrospray ionization-tandem MS (MS/MS) and MALDI-MS/MS and visualized using MALDI-IMS. SM species containing stearic were predominantly distributed in the loin and spinalis muscle, whereas SM species containing , lignoceric, and nervonic acids were predominantly distributed in transparent tissue. These results indicated that the distribution of SM species differed among the pork tissues, depending on the tissue-specific fatty composition. The total amount including all identified SM species was higher in the loin than in spinalis muscle. Pork is reportedly associated with increased risk for chronic diseases due to the high amount of heme iron. From the observation of color, the amount of heme iron was lower in loin than in spinalis muscle. Thus, the degree of risk for chronic diseases might be lower in the loin than in spinalis muscle. This is the first report on the tissue-specific distribution of SM species in meat at a microscopic resolution using IMS. MALDI-IMS analysis may be useful in assessing the association between SM species and quality parameters of pork meat. PRACTICAL APPLICATION: Sphingomyelin (SM) species are major sphingolipids in pork meat. SM species affect quality parameters such as health benefits due to their protective properties against colon cancer and atherosclerosis. Matrix-assisted laser desorption/ionization-imaging mass spectrometry analysis combined with liquid chromatography-electrospray ionization-tandem mass spectrometry is a suitable method to directly investigate the distribution and composition of SM species at microscopic level among different tissues of pork meat. Therefore, this method is useful to assess the SM species-induced health effect of different tissues of pork meat.© 2019 Institute of Food Technologists®.

Keyword: metabolism

Clinopodium chinense Attenuates -Induced Vascular Endothelial Inflammation and Insulin Resistance through TLR4-Mediated NF- B and MAPK .

Elevated (PA) levels are associated with the development of inflammation, insulin resistance (IR) and endothelial dysfunction. Clinopodium chinense (Benth.) O. Kuntze has been shown to lower blood glucose and attenuate high glucose-induced vascular endothelial cells injury. In the present study we investigated the effects of ethyl acetate extract of C. chinense (CCE) on PA-induced inflammation and IR in the vascular endothelium and its molecular mechanism. We found that CCE significantly inhibited PA-induced toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs). Consequently, this led to the inhibition of the following downstream adapted proteins myeloid differentiation primary response gene 88, Toll/interleukin-1 receptor domain-containing adaptor-inducing interferon- and TNF receptor-associated factor 6. Moreover, CCE inhibited the phosphorylation of Ikappa B kinase , nuclear factor kappa-B (NF- B), c-Jun N-terminal kinase, extracellular regulated protein kinases, p38-mitogen-activated protein kinase (MAPK) and subsequently suppressed the release of tumor necrosis factor- , interleukin-1 (IL-1 ) and IL-6. CCE also inhibited IRS-1 serine phosphorylation and ameliorated insulin-mediated tyrosine phosphorylation of IRS-1. Moreover, CCE restored serine/threonine kinase and endothelial nitric oxide synthase (eNOS) activation and thus increased insulin-mediated nitric oxide (NO) production in PA-treated HUVECs. This led to reverse insulin mediated endothelium-dependent relaxation, eNOS phosphorylation and NO production in PA-treated rat thoracic aortas. These results suggest that CCE can significantly inhibit the inflammatory response and alleviate impaired insulin signaling in the vascular endothelium by suppressing TLR4-mediated NF- B and MAPK . Therefore, CCE can be considered as a potential therapeutic candidate for endothelial dysfunction associated with IR and diabetes.

Keyword: metabolism

Tetrahydroxy stilbene glucoside alleviates -induced inflammation and apoptosis in cardiomyocytes by regulating miR-129-3p/Smad3 signaling.

Tetrahydroxy stilbene glucoside (TSG) has been reported to exert a cytoprotective effect against various toxicants. However, the function and mechanism of TSG in (PA)-induced inflammation and apoptosis in cardiomyocytes are still unknown. The present study was designed to investigate the post-transcriptional mechanism in TSG-treated cardiomyocytes\' inflammation and apoptosis induced by PA.The mRNA and protein levels were assayed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining was used to evaluate apoptosis using flow cytometry.TSG restricted the detrimental effects, including the activated inflammatory response and apoptosis, of PA in cardiomyocytes, as well as the up-regulation of miR-129-3p and down-regulation of p-Smad3 expression. In addition, bioinformatics and experimental analysis suggested that Smad3 was a direct target of miR-129-3p, which could inhibit or enhance the expression of p-Smad by transfection with miR-129-3p mimics or inhibitors, respectively. Furthermore, our results demonstrated that overexpression of Smad3 reversed the inhibition of inflammation and apoptosis by overexpression of miR-129-3p in PA-stimulated cardiomyocytes.TSG targeted to miR-129-3p/Smad3 signaling inhibited PA-induced inflammation and apoptosis in cardiomyocytes.

Keyword: metabolism

Free fatty -induced histone acetyltransferase activity accelerates lipid accumulation in HepG2 cells.

Non-alcoholic fatty liver disease (NAFLD) is a common disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and lipid in the liver have not been thoroughly investigated.Following oleic- (OPA)-induced lipid accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between lipid accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and lipid accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: metabolism

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines , and gut microbiome-derived . Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested fat decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: metabolism

Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured Hepatocytes.

A fluorescence microscopic method for characterizing size, quantity, and oxidation of lipid droplets (LDs) in HepG2 cells was developed. LDs were induced by (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and lipid peroxides. Each fatty increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2\'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.

Keyword: metabolism

Metabolomic Analysis of Membranous Glomerulonephritis: Identification of a\xa0Diagnostic Panel and Pathogenic .

Primary membranous glomerulonephritis (MGN) is a major cause of nephrotic syndrome in adults. Its diagnosis is based on invasive biopsy, and the current traditional serum or urinary biomarkers, such as the anti-phospholipase A2 receptor, are not adequately sensitive or specific.Our purpose is to identify a sensitive and specific noninvasive panel of biomarkers for the diagnosis of MGN by using metabolomic techniques and to explore the pathogenic that are involved in disease development.The urine metabolome of 66 MGN patients, 31 healthy controls, and 72 disease controls, were analyzed using nuclear magnetic resonance (NMR) and gas chromatography-tandem mass spectrometry (GC-MS/MS). Advanced multivariate statistical analyses were performed for the construction of diagnostic models and biomarker discovery. Receiver operating characteristic (ROC) curve analysis was used to suggest the most sensitive and specific diagnostic panel.The NMR-based diagnostic model showed allantoic and deoxyuridine as the most overrepresented and underrepresented biomarkers, respectively differentiating MGN from both control groups. The GC-MS/MS-based diagnostic model showed oxalic as the most overrepresented biomarker and 2-hydroxyglutaric lactone as the only underrepresented specific biomarker. A panel of a combination of the most accurate predictors of NMR and GC-MS/MS was composed of α-hydroxybutyric , 3,4-Dihydroxymandelic , 5α-cholestanone, 2-hydroxyglutaric lactone, nicotinamide, epicoprostanol, and . Nine impaired were identified in MGN, such as pyrimidine and NAD salvage.This comprehensive metabolomic study of MGN indicates a panel of promising biomarkers, which is complementary to current traditional biomarkers, and needs to be validated in a larger cohort.Copyright © 2019 IMSS. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

of 13C-Labeled Fatty Acids in Term Human Placental Explants by Liquid Chromatography-Mass Spectrometry.

Placental lipid transport and are poorly understood despite the importance for fetal development and lifelong health. We aimed to explore fatty (FA) processing in human villous placental explants from seven uncomplicated term singleton pregnancies delivered by elective cesarean section. Explants were treated with stable isotope-labeled (13C-PA), oleic (13C-OA), or docosahexaenoic (13C-DHA) for 3, 24, or 48 hours. Stable isotope-labeled lipids synthesized by placental explants from labeled FA were quantified, alongside endogenous unlabeled placental lipids, by liquid chromatography-mass spectrometry. Labeled phosphatidylcholines (PCs), triacylglycerols (TAGs), and phosphatidylethanolamines were detected in explants, whereas labeled lysophosphatidylcholines were found in both explants and conditioned media. 13C-PA was primarily directed into PC synthesis (74% of 13C-PA-labeled lipids), whereas 13C-OA was directed almost equally into PC and TAG synthesis (45% and 53%, respectively, of 13C-OA-labeled lipids). 13C-DHA was only detectable in TAGs. TAGs demonstrated the highest isotopic enrichment for all 13C-FAs with 13C-OA-TAGs comprising >50% of total OA-TAGs (unlabeled and labeled), consistent with TAGs being a labile and accessible reservoir for FA storage. Variations in lipid incorporation were correlated to maternal glycemia and body mass index, suggesting that this experimental model could be used to investigate the effect of maternal factors on placental lipid . We conclude that lipid partitioning of freshly imported FAs into labile and less labile lipid reservoirs in placenta is FA dependent. This process may partly mediate the physiological preferential transplacental transfer of particular FAs to the fetus, but may also be implicated in the fetoplacental pathophysiology of maternal dysfunction.Copyright © 2019 Endocrine Society.

Keyword: metabolism

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti-obesity remedy. Several pharmacological studies have shown its beneficial effects in . Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate lipid and glucose and are therapeutic targets for type 2 diabetes. This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, lipid accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased lipid accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: metabolism

Bioconversion of barley straw lignin into biodiesel using Rhodococcus sp. YHY01.

Rhodococcus sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize p-coumaric , cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38\u202fg dcw/L, 0.25\u202fg dcw/L and 0.1\u202fg dcw/L, and lipid accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC) value for p-coumaric (13.4\u202fmM), cresol (7.9\u202fmM), and 2,6 dimethoxyphenol (3.4\u202fmM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for Rhodococcus sp. YHY01 and resulted in 0.130\u202fg dcw/L with 39% w/w lipid accumulation. Major fatty acids were (C16:0) 51.87%, palmitoleic (C16:l) 14.90%, and oleic (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: metabolism

Chemical composition, in vitro gas production, methane production and fatty profile of canola silage (Brassica napus) with four levels of molasses.

The objective of this study was to investigate the effect of four levels of molasses on chemical composition, in vitro digestibility, methane production and fatty profile of canola silages. A canola (Brassica napus var. Monty) crop was established in a small-scale agricultural farm and harvested 148\xa0days after sowing. Four levels of molasses were tested with respect to the fresh weight (1.5\xa0kg); these were 1% (CS-1), 2% (CS-2), 3% (CS-3) and 4% (CS-4) molasses, and 0% molasses (CS-0) was included as a control. A total of 45 microsilages were prepared using PVC pipes (4\xa0in. of diameter × 20\xa0cm of length), and the forage was compressed using a manual press. The effects of control and treatments were tested using the general linear model Y\u2009=\u2009μ\u2009+\u2009T\u2009+\u2009E. The linolenic (C18:3n3), (C16:0) and linoleic methyl ester (C18:2n6c) accounted for 30%, 21% and 10.5% of total fatty acids, respectively; the fermentation parameters and in vitro methane production were not affected (P\u2009>\u20090.05) by treatments; in vitro digestibility decreased significantly (P\u2009<\u20090.05) as the level of molasses increased. It was concluded that CS-4 improved the DM content by 9% and showed high content of linolenic methyl ester. The gross energy of canola silages could favour the oleic methyl ester.

Keyword: metabolism

Benefits of sea buckthorn (Hippophae rhamnoides) pulp oil-based mouthwash on oral health.

The purpose of this study was to conduct phytochemical analysis of sea buckthorn pulp oil and to evaluate the antimicrobial, anti-biofilm and antioxidant activities of its mouthwash form.Fatty composition of the sea buckthorn pulp oil was determined by GC-MS analysis, which revealed that, mono-unsaturated fatty , palmitoleic and saturated fatty , , were the major constituents. The antimicrobial and the anti-biofilm capacities of sea buckthorn pulp oil mouthwash form were evaluated against Streptococcus gordonii, Porphyromonas gingivalis, Actinomyces viscosus and Candida albicans, according to the European Norms, and the Biofilm Ring Test , respectively. These activities were then compared with those of chlorhexidine and herbal mouthwashes. The sea buckthorn-based mouthwash was bactericidal against S. gordonii and P. gingivalis, bacteriostatic against A. viscosus and showed no antifungal effect. Regardless of the strains used, complete inhibition of biofilm formation was achieved. The antioxidant activity of this experimental mouthwash was also assessed by DPPH and NBT assays.Sea buckthorn mouthwash showed anti-biofilm activities against select single and multiple oral bacterial species.In this study, a mouthwash derived from sea buckthorn (Hippophae rhamnoides) pulp oil has been experimented, for the first time, in order to overcome the problem of a large number of available synthetic mouthwashes which have side effects on teeth, gums and mucous membranes. This mouthwash seemed to be a suitable alternative for a preventive agent for periodontal inflammation.© 2019 The Society for Applied Microbiology.

Keyword: metabolism

Hsp74/14-3-3σ Complex Mediates Centrosome Amplification by High Glucose, Insulin, and .

It has been reported recently that type 2 diabetes promotes centrosome amplification via 14-3-3σ/ROCK1 complex. In the present study, 14-3-3σ interacting proteins are characterized and their roles in the centrosome amplification by high glucose, insulin, and are investigated. Co-immunoprecipitation in combination with MS analysis identified 134 proteins that interact with 14-3-3σ, which include heat shock 70 kDa protein 4 (Hsp74). Gene ontology analyses reveal that many of them are enriched in binding activity. Kyoto Encyclopedia of Genes and Genomes analysis shows that the top three enriched are ribosome, carbon , and biosynthesis of amino acids. Molecular and functional investigations show that the high glucose, insulin, and increase the expression and binding of 14-3-3σ and Hsp74 as well as centrosome amplification, all of which are inhibited by knockdown of 14-3-3σ or Hsp74. Moreover, molecular docking analysis shows that the interaction between the 14-3-3σ and the Hsp74 is mainly through hydrophobic contacts and a lesser degree ionic interactions and hydrogen bond by different amino acids residues. In conclusion, the results suggest that the experimental treatment triggers centrosome amplification via upregulations of expression and binding of 14-3-3σ and Hsp74.© 2019 The Authors. Proteomics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: metabolism

Removal of phthalic esters from sea buckthorn (Hippophae rhamnoides L.) pulp oil by steam distillation and molecular distillation.

Sea buckthorn (Hippophae rhamnoides L.) pulp oils (SPOs) are rich in a variety of beneficial bioactive ingredients. Nevertheless, SPOs would be exposed to plastic equipment during processing, resulted in increasing phthalates contents and edible risk, as well as affecting oil quality. For these reasons, the effects of two stages steam distillation (SD2) and two stages molecular distillation (MD2) on phthalic esters (PAEs) content were investigated and compared in the present work. Compared with SD2, MD2 showed higher removal rates of seven selected PAEs from the SPO. Even if the initial concentration of DBP and DEHP in R-SPO were 1.626 and 10.933\u202fmg/kg respectively, the concentration of DBP and DINP could be reduced below the limit set by China government after treated with MD2. Besides that, there was no trans-fatty acids generated in SPO during the distillation process.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

Obesity increases the risk of developing diabetes mellitus. Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in diabetes with obesity. Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II diabetes. Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling , including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of diabetes with obesity that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for diabetes with obesity.

Keyword: metabolism

3--Methyl-Alkylgallates Inhibit Fatty Desaturation in Mycobacterium tuberculosis.

In the quest for new antibacterial lead structures, activity screening against identified antitubercular effects of gallic derivatives isolated from the Nigerian mistletoe Structure-activity relationship studies indicated that 3--methyl-alkylgallates comprising aliphatic ester chains with four to eight carbon atoms showed the strongest growth inhibition against , with a MIC of 6.25\u2009μM. Furthermore, the most active compounds (3--methyl-butyl-, 3--methyl-hexylgallate, and 3--methyl-octylgallate) were devoid of cytotoxicity against various human cell lines. Furthermore, 3--methyl-butylgallate showed favorable absorption, distribution, , and excretion (ADME) criteria, with a of 6.2\u2009×\u200910cm/s, and it did not inhibit P-glycoprotein (P-gp), CYP1A2, CYP2B6 or CYP3A4. Whole-genome sequencing of spontaneous resistant mutants indicated that the compounds target the stearoyl-coenzyme A (stearoyl-CoA) delta-9 desaturase DesA3 and thereby inhibit oleic synthesis. Supplementation assays demonstrated that oleic addition to the culture medium antagonizes the inhibitory properties of gallic derivatives and that sodium salts of saturated and stearic did not show compensatory effects. The moderate bactericidal effect of 3--methyl-butylgallate in monotreatment was synergistically enhanced in combination treatment with isoniazid, leading to sterilization in liquid culture.Copyright © 2019 American Society for Microbiology.

Keyword: metabolism

Metabolomic change due to combined treatment with myo-inositol, D-chiro-inositol and glucomannan in polycystic ovarian syndrome patients: a pilot study.

Polycystic ovarian syndrome (PCOS) is a highly variable syndrome and one of the most common female endocrine disorders. Although the association inositols-glucomannan may represent a good therapeutic strategy in the treatment of PCOS women with insulin resistance, the effect of inositols on the metabolomic profile of these women has not been described yet.Fifteen PCOS-patients and 15 controls were enrolled. Patients were treated with myo-inositol (1.75\u2009g/day), D-chiro-inositol (0.25\u2009g/day) and glucomannan (4\u2009g/day) for 3\u2009months. Blood concentrations of glucose, insulin, triglycerides and cholesterol, and ovary volumes and antral follicles count, as well as metabolomic profiles, were evaluated for control subjects and for cases before and after treatment. PCOS-patients had higher BMI compared with Controls, BMI decreased significantly after 3\u2009months of treatment although it remained significantly higher compared to controls. 3-methyl-1-hydroxybutyl-thiamine-diphosphate, valine, phenylalanine, ketoisocapric, linoleic, lactic, glyceric, citric and , glucose, glutamine, creatinine, arginine, choline and tocopherol emerged as the most relevant metabolites for distinguishing cases from controls.Our pilot study has identified a complex network of serum molecules that appear to be correlated with PCOS, and with a combined treatment with inositols and glucomannan.ClinicalTial.gov, . Registered 1st August 2018 - Retrospectively registered, .

Keyword: metabolism

Proteomic profiling and integrated analysis with transcriptomic data bring new insights in the stress responses of after an arrest during high-temperature ethanol fermentation.

The thermotolerant yeast is a potential candidate for high-temperature fermentation. When was used for high-temperature ethanol fermentation, a fermentation arrest was observed during the late fermentation stage and the stress responses have been investigated based on the integration of RNA-Seq and metabolite data. In order to bring new insights into the cellular responses of after the fermentation arrest during high-temperature ethanol fermentation, quantitative proteomic profiling and integrated analysis with transcriptomic data were performed in this study.Samples collected at 14, 16, 18, 20 and 22\xa0h during high-temperature fermentation were subjected to isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic profiling and integrated analysis with transcriptomic data. The correlations between transcripts and proteins for the comparative group 16\xa0h vs 14\xa0h accounted for only 4.20% quantified proteins and 3.23% differentially expressed proteins\xa0(DEPs), respectively, much higher percentages of correlations (30.56%-59.11%) were found for other comparative groups (i.e., 18\xa0h vs 14\xa0h, 20\xa0h vs 14\xa0h, and 22\xa0h vs 14\xa0h). According to Spearman correlation tests between transcriptome and proteome (the absolute value of a correlation coefficient between 0.5 and 1 indicates a strong correlation), poor correlations were found for all quantified proteins ( = -\u20090.0355 to 0.0138), DEPs ( = -\u20090.0079 to 0.0233) and the DEPs with opposite expression trends to corresponding differentially expressed genes (DEGs) ( = -\u20090.0478 to 0.0636), whereas stronger correlations were observed in terms of the DEPs with the same expression trends as the correlated DEGs ( = 0.5593 to 0.7080). The results of multiple reaction monitoring (MRM) verification indicate that the iTRAQ results were reliable. After the fermentation arrest, a number of proteins involved in transcription, translation, oxidative phosphorylation and fatty were down-regulated, some molecular chaperones and proteasome proteins were up-regulated, the ATPase activity significantly decreased, and the total fatty acids gradually accumulated. In addition, the contents of , oleic , C16, C18, C22 and C24 fatty acids increased by 16.77%, 28.49%, 14.14%, 26.88%, 628.57% and 125.29%, respectively.This study confirmed some biochemical and enzymatic alterations provoked by the stress conditions in the specific case of : such as decreases in transcription, translation and oxidative phosphorylation, alterations in cellular fatty composition, and increases in the abundance of molecular chaperones and proteasome proteins. These findings provide potential targets for further engineering towards improvement of the stress tolerance in .

Keyword: metabolism

Multi-Functional Desaturases in Two Spodoptera Moths with ∆11 and ∆12 Desaturation Activities.

The beet armyworm, Spodoptera exigua, uses (Z,E)-9,12-tetradecadienyl acetate as the major component of its sex pheromone. Previous isotope-labeling experiments demonstrated an unusual ∆12 desaturase activity involved in the biosynthesis of this compound; however, the putative ∆12 desaturase gene has not been reported to date. In the present study, we confirmed this ∆12 desaturation pathway by in vivo labeling experiments, and characterized candidate desaturase genes in a yeast heterologous expression system. We demonstrated that a pheromone gland-specific desaturase, SexiDes5, uses and the subsequently chain-shortened product (Z)-9-tetradecenoic as substrates to produce (Z)-11-hexadecenoic and (Z,E)-9,12-tetradecadienoic acids, respectively. In addition, the homologous desaturase SlitDes5 from the congeneric Spodoptera litura had analogous functions.

Keyword: metabolism

Transcriptome analysis of elaiosomes and seeds provide insights into fatty biosynthesis.

Elaiosomes are specialized fleshy and edible seed appendages dispersed by ants. Lipids are the primary components of elaiosomes. is a well-known plant, the seeds of which are dispersed by ants. Previous studies have identified the presence of primary fatty acids in its elaiosomes and seeds. However, the molecular mechanisms underlying fatty biosynthesis in elaiosomes remain unknown.In order to gain a comprehensive transcriptional profile of the elaiosomes and seeds of , and understand the expression patterns of genes associated with fatty biosynthesis, four different developmental stages, including the flower-bud (Ch01), flowering (Ch02), young seed (Ch03), and mature seed (Ch04) stages, were chosen to perform whole-transcriptome profiling through the RNA-seq technology (Illumina NGS sequencing).A total of 63,064 unigenes were generated from 12 libraries. Of these, 7,323, 258, and 11,540 unigenes were annotated with 25 Cluster of Orthologous Groups, 43 Gene Ontology terms, and 373 Kyoto Encyclopedia of Genes and Genomes , respectively. In addition, 322 genes were involved in lipid transport and , and 508 genes were involved in the lipid . A total of 41 significantly differentially expressed genes (DEGs) involved in the lipid were identified, most of which were upregulated in Ch03 compared to Ch02, indicating that fatty biosynthesis primarily occurs during the flowering to the young seed stages. Of the DEGs, acyl-ACP thioesterases, acyl carrier protein desaturase (), and malonyl CoA-ACP transacylase were involved in synthesis; stearoyl-CoA desaturase and were involved in oleic synthesis, and acyl-lipid omega-6 desaturase was involved in linoleic synthesis.

Keyword: metabolism

PAQR3 regulates phosphorylation of FoxO1 in insulin-resistant HepG2 cells via NF-κB signaling pathway.

Insulin resistance is a significant feature of type 2 diabetes mellitus and glucose and lipid disorders. Activation of NF-κB signaling pathway plays an important role in the formation of insulin resistance. FoxO1 plays a major role in regulating glucose and lipid , as well as insulin signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under insulin resistance. In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced insulin-resistant HepG2 cells, thereby causing glucose and lipid disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced insulin-resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and lipid disorders induced by insulin resistance.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Integration of proteomics and metabolomics data of early and middle season Hass avocados under heat treatment.

Ripening heterogeneity of Hass avocados results in inconsistent quality fruit delivered to the triggered and ready to eat markets. This research aimed to understand the effect of a heat shock (HS) prior to controlled atmosphere (CA) storage on the reduction of ripening heterogeneity. HS prior to CA storage reduces more drastically the ripening heterogeneity in middle season fruit. Via correlation network analysis we show the different metabolomics between HS and CA. High throughput proteomics revealed 135 differentially expressed proteins unique to middle season fruit triggered by HS. Further integration of metabolomics and proteomics data revealed that HS reduced the glycolytic throughput and induced protein degradation to deliver energy for the alternative ripening . l-isoleucine, l-valine, l-aspartic and ubiquitin carboxyl-terminal hydrolase involved in protein degradation were positively correlated to HS samples. Our study provides new insights into the effectiveness of HS in synchronizing ripening of Hass avocados.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial energy supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in lipid during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin . We further demonstrated the impact of cellular lipid , and particularly cardiolipin , during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: metabolism

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: metabolism

Uncommon properties of lipid biosynthesis of isolated plastids in the unicellular red alga .

Red algae are a large group of photosynthetic eukaryotes that diverged from green algae over one billion years ago, and have various traits distinct from those of both green algae and land plants. Although most red algae are marine species (both unicellular and macrophytic), the Cyanidiales class of red algae includes unicellular species which live in hot springs, such as , which is a model species for biochemical and molecular biological studies. Lipid in red algae has previously been studied in intact cells. Here, we present the results of radiolabeling and stable isotope labeling experiments in intact plastids isolated from the unicellular red alga . We focused on two uncommon features: First, the galactose moiety of monogalactosyldiacylglycerol was efficiently labeled with bicarbonate, indicating that an unknown pathway for providing UDP-galactose exists within the plastid. Second, saturated fatty acids, namely, and stearic acids, were the sole products of fatty synthesis in the plastid, and they were efficiently exported. This finding suggests that the endoplasmic reticulum is the sole site of desaturation. We present a general principle of red algal lipid biosynthesis, namely, \'indigenous C18 fatty acids are neither desaturated nor directly utilized within the plastid\'. We believe that this is valid in both lacking polyunsaturated fatty acids and marine red algae with a high content of arachidonic and eicosapentaenoic acids.

Keyword: metabolism

Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk fat content (3.60 vs. 3.41%), milk fat yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% fat-corrected milk (47.6 vs. 46.5 kg/d), and energy-corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk fat yield (1.70 vs. 1.66 kg/d), and 3.5% fat-corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk fat yield, energy-corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose in the obese mice.

Keyword: metabolism

Mp1 Protein, a Novel Virulence Factor, Carries Two Arachidonic -Binding Domains To Suppress Inflammatory Responses in Hosts.

infection causes talaromycosis (previously known as penicilliosis), a very important opportunistic systematic mycosis in immunocompromised patients. Different virulence mechanisms in have been proposed and investigated. In the sera of patients with talaromycosis, Mp1 protein (Mp1p), a secretory galactomannoprotein antigen with two tandem ligand-binding domains (Mp1p-LBD1 and Mp1p-LBD2), was found to be abundant. Mp1p-LBD2 was reported to possess a hydrophobic cavity to bind copurified (PLM). It was hypothesized that capturing of lipids from human hosts by expressing a large quantity of Mp1p is a virulence mechanism of It was shown that expression of Mp1p enhanced the intracellular survival of by suppressing proinflammatory responses. Mechanistic study of Mp1p-LBD2 suggested that arachidonic (AA), a precursor of paracrine signaling molecules for regulation of inflammatory responses, is the major physiological target of Mp1p-LBD2. In this study, we use crystallographic and biochemical techniques to further demonstrate that Mp1p-LBD1, the previously unsolved first lipid binding domain of Mp1p, is also a strong AA-binding domain in Mp1p. These studies on Mp1p-LBD1 support the idea that the highly expressed Mp1p is an effective AA-capturing protein. Each Mp1p can bind up to 4 AA molecules. The crystal structure of Mp1p-LBD1-LBD2 has also been solved, showing that both LBDs are likely to function independently with a flexible linker between them. and potentially other pathogens highly expressing and secreting proteins similar to Mp1p can severely disturb host signaling cascades during proinflammatory responses by reducing the availabilities of important paracrine signaling molecules.Copyright © 2019 American Society for Microbiology.

Keyword: metabolism

Risk of diabetes associated with fatty acids in the de novo lipogenesis pathway is independent of insulin sensitivity and response: the Insulin Resistance Atherosclerosis Study (IRAS).

To examine the associations of fatty acids in the de novo lipogenesis (DNL) pathway, specifically myristic (14:0), (16:0), palmitoleic (c16:1\u2009n-7), myristoleic (c14:1n5), stearic (18:0) and oleic (c18:1\u2009n-9), with 5-year risk of type 2 diabetes. We hypothesized that DNL fatty acids are associated with risk of type 2 diabetes independent of insulin sensitivity.We evaluated 719 (mean age 55.1±8.5 years, 44.2% men, 42.3% Caucasians) participants from the Insulin Resistance Atherosclerosis Study. Multivariable logistic regression models with and without adjustment of insulin sensitivity were used to assess prospective associations of DNL fatty acids with incident type 2 diabetes.Type 2 diabetes incidence was 20.3% over 5\u2009years. In multivariable regression models, , palmitoleic, myristic, myristoleic and oleic acids were associated with increased risk of type 2 diabetes (p<0.05). had the strongest association (OR per standard unit of 1.46; 95%\u2009CI 1.23 to 1.76; p<0.001), which remained similar with addition of insulin sensitivity and acute insulin response (AIR) to the model (OR 1.36; 95%\u2009CI 1.09 to 1.70, p=0.01). Oleic and palmitoleic acids were also independently associated with incident type 2 diabetes. In multivariable models, ratios of fatty acids corresponding to stearoyl CoA desaturase-1 and Elovl6 enzymatic activity were significantly associated with risk of type 2 diabetes independent of insulin sensitivity and AIR.We observed associations of DNL fatty acids with type 2 diabetes incidence independent of insulin sensitivity.

Keyword: metabolism

N-3 Polyunsaturated Fatty Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Diabetes Rats by Postponing Shortening of Hepatic Telomeres and Modulating Liver .

The long-term influence of gestational diabetes mellitus (GDM) on offspring and the effect of omega-3 polyunsaturated fatty acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and inflammation in the liver of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the liver, in which numerous metabolites and were altered when GDM offspring grew to old age. Many metabolites were related to diabetes risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious diabetes risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: metabolism

Berberine (BBR) Attenuated (PA)-Induced Lipotoxicity in Human HK-2 Cells by Promoting Peroxisome Proliferator-Activated Receptor α (PPAR-α).

BACKGROUND Berberine (BBR), a natural alkaloid isolated from Coptis chinensis, has frequently been reported as an antidiabetic reagent, partly due to its lipid-lowering activity. Evidence suggests that BBR ameliorates palmitate-induced lipid deposition and apoptosis in renal tubular epithelial cells (TECs), which tracks in tandem with the enhancement of peroxisome proliferator-activated receptor alpha (PPAR-alpha). The study aim was to investigate the roles of BBR in renal lipotoxicity in vitro, and investigate whether PPAR-alpha was the underlying mechanism. MATERIAL AND METHODS Human TECs (HK-2 cells) were injured with (PA), and then treated with BBR, BBR+PPAR-alpha inhibitor (GW6471), and PA+PPAR-alpha agonist (fenofibrate). Endoplasmic reticulum (ER) stress was assessed by measuring the expression of prospective evaluation of radial keratotomy (PERK), C/EBP-homologous protein (CHOP), and 78 kDa glucose-regulated protein (GRP78). Lipid was assessed by determining lipid anabolism-associated genes, including fatty synthase (FAS), acetyl-CoA carboxylase (ACC), and lipoprotein lipase (LPL), as well as lipid catabolism-associated gene, including carnitine palmitoyl transferase 1 (CPT1). Inflammatory response of HK-2 cells was evaluated by measuring interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha. Cell apoptosis and protein levels of cleaved-caspase-3 were evaluated. RESULTS PA downregulated PPAR-alpha and induced server lipotoxicity in HK-2 cells by ER stress, increasing lipid deposition, and elevating inflammatory response of HK-2 cells accompanied with inducting cell apoptosis and cleaved-caspase-3, which were obviously reversed by additional treatment of BBR or PPAR-alpha agonist. However, the protective effect of BBR in PA-induced lipotoxicity in HK-2 cells was significantly ameliorated by PPAR-alpha inhibitor. CONCLUSIONS BBR attenuated PA-induced lipotoxicity via the PPAR-alpha pathway.

Keyword: metabolism

Molecular interplay between microRNA-130a and PTEN in -mediated impaired function of endothelial progenitor cells: Effects of metformin.

Metformin serves an important role in improving the functions of endothelial progenitor cells (EPCs). MicroRNAs (miRNAs), small non‑coding RNAs, have been investigated as significant regulators of EPC vascular functions. The present study investigated the molecular crosstalk between metformin and miRNA‑130a (miR‑130a) in the functions of EPCs exposed to (PA). Isolated EPCs were treated with metformin, PA, and metformin + PA, respectively. Cell Counting Kit‑8, Transwell and Matrigel assays were performed to detect the proliferation, migration and tube formation ability of EPCs following different treatments. The expression of miR‑130a, phosphatase and tensin homolog (PTEN) and phosphorylated‑AKT was analyzed by reverse transcription‑quantitative polymerase chain reaction and western blotting. The specific mechanism underlying the function of metformin in EPCs was further elucidated by transfecting miR‑130a mimics and inhibitor to overexpress and inhibit the expression of miR‑130a in EPCs, respectively. EPCs exhibited impaired functions of proliferation (P<0.01 compared with the control), migration (P<0.01 compared with the control) and tube formation (P<0.01 compared with the control) following treatment with PA, and the expression levels of miR‑130a and PTEN were decreased and increased, respectively. However, the presence of metformin, or the overexpression of miR‑130a using miR‑130a mimic alleviated the impairment of angiogenesis and proliferation, decreased the expression of PTEN and activated the phosphoinositide‑3 kinase/AKT pathway in EPCs exposed to PA. By contrast, downregulating the expression of miR‑130a with a miR‑130a inhibitor reversed the metformin‑mediated protection. These results demonstrate the beneficial effect of miR‑130a/PTEN on EPC functions, which can be regulated by metformin. The effects of metformin on improving PA‑induced EPC dysfunction are mediated by miR‑130a and PTEN, which may assist in the prevention and/or treatment of diabetic vascular disease.

Keyword: metabolism

Acanthoic modulates lipogenesis in nonalcoholic fatty liver disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic fatty liver disease (NAFLD) associated with lipid accumulation by activating Farnesoid X receptor (FXR) and liver X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high-fat (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on lipid accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, liver index, and liver lipid droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated lipid accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating lipid accumulation and fatty synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolism

pH effects on the lipid and fatty acids accumulation in Chlamydomonas reinhardtii.

pH variations influence the delivery of essential nutrients and CO solubility, which impact algae . In this study the microalgal growth and chlorophyll, lipid, and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. Lipid content increased 3.0 (14.81% of dry cell weight, dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for ; 17.3, 14.7, and 25.7% for oleic ; and 9.8, 12.1, and 4.6% for linoleic ; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of lipid biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and lipid accumulation.© 2019 American Institute of Chemical Engineers.

Keyword: metabolism

d- chiro-Inositol Ameliorates High Fat Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high fat diet and saturated -treated hepatocytes. DCI attenuated free fatty uptake by the liver via lipid trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces lipid deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: metabolism

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial effects.

Keyword: metabolism

In vivo kinetic study of materno-fetal fatty transfer in obese and normal weight pregnant women.

Placental structure and function can be modified as a result of maternal obesity affecting materno-fetal fatty acids (FA) transport. We report for the first time, in humans and in vivo, the kinetics of placental FA transfer in normo-weight and in normolipemic obese pregnant women using stable isotopes. The administration of different tracer FA with similar behaviour to the mother at different time points allows the collection of kinetic information on materno-fetal transfer of FA despite only one sample of placenta and cord can be collected per subject. Computational modelling showed a good fit to the data when considering all maternal plasma lipid classes but not when based only on non-esterified FA. The novel approach using multiple tracer FA administration combined with computational modelling shows a consistent time course of placental tracer FA and predicted total FA accumulation.We analyse for the first time the in vivo materno-fetal kinetic transfer of fatty acids (FA) labelled with stable isotopes in control and obese (OB) pregnant women. Labelled FA with a similar (stearic : C-SA; : C-PA; oleic : C-OA) were orally administered at -4\xa0h, -8\xa0h and -12\xa0h, respectively prior to elective caesarean section to 10 pregnant women with a body mass index >30 (OB) and 10 with a body mass index in the range 20-25 (NW). Placenta, venous and arterial cord blood were collected obtaining a wide range of FA enrichments. A combined experimental and computational modelling analysis was applied. FA fractional synthesis rate (FSR) in placenta was 11-12%\xa0h . No differences were observed between NW and normo-lipidemic OB. It was not possible to estimate FA FSR in cord blood with this oral bolus dose approach. Computational modelling demonstrated a good fit to the data when all maternal plasma lipid classes were included but not with modelling based only on the non-esterified FA fraction. The estimated materno-fetal C-FA transfer was ∼1%. In conclusion, our approach using multiple C-FA tracers allowed us to estimated FSR in placental/maternal plasma but not in fetal/maternal compartments. Computational modelling showed a consistent time course of placental C-FA transfer and predicted total fetal FA accumulation during the experiment. We conclude that, in addition to non-esterified FA fraction in the maternal circulation, maternal plasma very low-density lipoprotein and other lipoproteins are important contributors to placental FA transfer to the fetus.© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.

Keyword: metabolism

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid , Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free fatty (FFA)- and oleic -and high-fat diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice liver were measured, and the expression levels of genes associated with lipogenesis, fatty oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with fatty oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of lipid , inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: metabolism

PAHSAs attenuate immune responses and promote β cell survival in autoimmune diabetic mice.

esters of hydroxy stearic acids (PAHSAs) are endogenous antidiabetic and antiinflammatory lipids. Here, we show that PAHSAs protect against type 1 diabetes (T1D) and promote β cell survival and function. Daily oral PAHSA administration to nonobese diabetic (NOD) mice delayed the onset of T1D and markedly reduced the incidence of T1D, whether PAHSAs were started before or after insulitis was established. PAHSAs reduced T and B cell infiltration and CD4+ and CD8+ T cell activation, while increasing Treg activation in pancreata of NOD mice. PAHSAs promoted β cell proliferation in both NOD mice and MIN6 cells and increased the number of β cells in NOD mice. PAHSAs attenuated cytokine-induced apoptotic and necrotic β cell death and increased β cell viability. The mechanism appears to involve a reduction of ER stress and MAPK signaling, since PAHSAs lowered ER stress in NOD mice, suppressed thapsigargin-induced PARP cleavage in human islets, and attenuated ERK1/2 and JNK1/2 activation in MIN6 cells. This appeared to be mediated in part by glucagon-like peptide 1 receptor (GLP-1R) and not the G protein-coupled receptor GPR40. PAHSAs also prevented impairment of glucose-stimulated insulin secretion and improved glucose tolerance in NOD mice. Thus, PAHSAs delayed the onset of T1D and reduced its incidence by attenuating immune responses and exerting direct protective effects on β cell survival and function.

Keyword: metabolism

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty . Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains plasticity in fatty desaturation and constitutes an unexplored rewiring in cancers.

Keyword: metabolism

Dynamics of Individual Fatty Acids in Muscle Fat Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C to C diet shift experiment and compound-specific δC analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and lipid droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic (LA; 18:2n6) and (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic (20:4n6) and docosahexaenoic (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular lipid droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in energy . The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects energy of birds during migration.

Keyword: metabolism

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) . However, whether TMAO aggravates liver steatosis by modulating BA and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA . Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: metabolism

Metabolomic analysis of significant changes in Lactobacillus casei Zhang during culturing to generation 4,000 under conditions of glucose restriction.

Lactic bacteria are being consumed more frequently as awareness of their health benefits has increased. The industrial production of lactic bacteria requires a comprehensive understanding of their survival stress, especially regarding changes in substances in a glucose-limited environment. In the present study, a metabolomic approach was applied to investigate Lactobacillus casei Zhang using cultures from a common ancestor that were permitted to evolve under conditions with normal or glucose-restricted media for up to 4,000 generations. Metabolomic analyses of intracellular and extracellular differential metabolites under De Man, Rogosa and Sharpe broth (2% vol/vol glucose; Oxoid Ltd., Basingstoke, UK) and glucose-restricted (0.02% vol/vol glucose in De Man, Rogosa and Sharpe broth) conditions were performed at generations 0, 2,000, and 4,000 and revealed 23 different metabolites. Myristic , ergothioneine, Lys-Thr, and palmitamide contents exhibited significant reductions between 0 and 4,000 generations, whereas nicotinate, histidine, , l-lysine, urocanate, thymine, and other substances increased. The dynamics of the involved in AA , including glycine, serine, and threonine , histidine , lysine degradation, and arginine and proline , were also a focus of the present study. There were also changes in several other , including vitamin B, thiamine, nicotinate, and nicotinamide, according to generation time. Additionally, in the present study we screened for key metabolites involved in the glucose-restricted response and provided a theoretical basis for comprehensively revealing the regulatory mechanisms associated with L. casei Zhang glucose restriction at the level. These findings also provide novel ideas and methods for analyzing the glucose-restricted stress response at the level.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: metabolism

Metabotropic Glutamate Receptor 5 and 8 Modulate the Ameliorative Effect of Ultramicronized Palmitoylethanolamide on Cognitive Decline Associated with Neuropathic Pain.

This study investigated whether metabotropic glutamate receptor (mGluR) 5 and 8 are involved in the effect of ultramicronizedpalmitoylethanolamide (um-PEA) on the cognitive behavior and long term potentiation (LTP) at entorhinal cortex (LEC)-dentate gyrus (DG) pathway in mice rendered neuropathic by the spare nerve injury (SNI). SNI reduced discriminative memory and LTP. Um-PEA treatment started after the development of neuropathic pain had no effects in sham mice, whereas it restored cognitive behavior and LTP in SNI mice. 2-Methyl-6-(phenylethynyl) pyridine (MPEP), a selective mGluR5 antagonist, improved cognition in SNI mice and produced a chemical long term depression of the field excitatory postsynaptic potentials (fEPSPs) in sham and SNI mice. After theta burst stimulation (TBS) MPEP restored LTP in SNI mice. In combination with PEA, MPEP antagonized the PEA effect on discriminative memory and decreased LTP in SNI mice. The (RS)-4-(1-amino-1-carboxyethyl)phthalic (MDCPG), a selective mGluR8 antagonist, did not affect discriminative memory, but it induced a chemical LTP and prevented the enhancement of fEPSPs after TBS in SNI mice which were treated or not treated with PEA. The effect of PEA on LTP and cognitive behavior was modulated by mGluR5 and mGluR8. In particular in the SNI conditions, the mGluR5 blockade facilitated memory and LTP, but prevented the beneficial effects of PEA on discriminative memory while the mGluR8 blockade, which was ineffective in itself, prevented the favorable action of the PEA on LTP. Thus, although their opposite roles (excitatory/inhibitory of the two receptor subtypes on the glutamatergic system), they appeared to be required for the neuroprotective effect of PEA in conditions of neuropathic pain.

Keyword: metabolism

CD36 mediates palmitate -induced metastasis of gastric cancer via AKT/GSK-3β/β-catenin pathway.

Gastric cancer (GC) has a clear predilection for metastasis toward the omentum which is primarily composed of adipose tissue, indicating that fatty acids may contribute to this phenomenon. However their function remains poorly understood in GC. In this study, we investigated the role of palmitate (PA) and its cellular receptor CD36 in the progression of GC.Immunohistochemical (IHC) staining was performed to detect CD36 expression in GC tissues and its clinical significance was determined statistically. CD36 over-expression and knock-down expression cell models were developed and tested in vitro. Wound-healing assays, migration assays, and invasion assays were performed and peritoneal implants into nude mice were done to assess the biological effects of PA and CD36. The underlying mechanisms were investigated using western blot, immunofluorescence (IF), quantitative real-time PCR (qRT-PCR) and antibody blocking assays.PA promoted the metastasis of GC by phosphorylation of AKT, which facilitated the nuclear localization of β-catenin through inactivation of GSK-3β via phosphorylation. This tumor-promoting effect of PA was mediated by CD36, a cell surface receptor of fatty acids (FAs). The higher the CD36 expression levels in GC tissues correlated with the poorer the prognosis of patients according to the TCGA database, the GEO database and our own clinical data.Our experiments established CD36 as a key mediator of FA-induced metastasis of GC via the AKT/GSK-3β/β-catenin signaling pathway. CD36 might, therefore, constitute a potential therapeutic target for clinical intervention in GC.

Keyword: metabolism

κ-Opioid receptor stimulation reduces palmitate-induced apoptosis via Akt/eNOS signaling pathway.

This study was designed to test the hypothesis that κ-opioid receptor (κ-OR) stimulation reduces palmitate-induced HUVECs apoptosis and to investigate its mechanisms.HUVECs were subjected to sodium palmitate, apoptosis and cell viability were determined, HUVECs were treated with specific inhibitors to PI3K, Akt, eNOS and siRNAs targeting κ-OR and Akt. Groups were divided as follows: the control group, the sodium palmitate group, the sodium palmitate+U50,488H (a selective κ-OR agonist) group and the sodium palmitate+U50,488H\u2009+\u2009nor-BNI (a selective κ-OR antagonist) group.Treatment with sodium palmitate significantly reduced cell viability and increased apoptosis rate which were significantly alleviated by pretreatment with U50,488H, the effect of U50,488H was abolished by nor-BNI. Phosphorylation of Akt and eNOS, as well as NO production were attenuated and accompanied by an increased expression of caspase 3 when HUVECs were subjected to sodium palmitate, and all these changes were restored by pretreatment with U50,488H, the effects of U50,488H were abolished by nor-BNI, and specific inhibitors to PI3K, Akt, eNOS, respectively. SiRNAs targeting κ-OR or Akt abolished the effects of U50,488H on phosphorylation of Akt and eNOS as well as the expressions of caspase 3, Bax and Bcl-2. SiRNAs targeting Akt elicited no effect on the expression of κ-OR.This study provides the evidence for the first time that κ-OR stimulation possesses anti-palmitate-induced apoptosis effect, which is mediated by PI3K/Akt/eNOS signaling pathway.

Keyword: metabolism

Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering hyperlipidemia via regulating the AMPK/SREBP1/PPARα signaling cascade of lipid .

The present study for the first time aims to examine the hypothesis that circulating gamma-glutamyl carboxylated growth arrest specific protein 6 (Gla-Gas6) deficiency may be associated with hyperlipidemia and vitamin K (VK) supplementation may ameliorate the impaired lipid homeostasis via activating Gas6 protein. Subjects with hyperlipidemia (n=22) and age-matched healthy controls (n=19) were included in this study. Results showed that plasma levels of Gla-Gas6 protein and VK were significantly lower in hyperlipidemic subjects compared to control. Moreover, Gla-Gas6 levels were significantly and positively correlated with VK (P=.034, r=0.452) and negatively with triglyceride (P=.022, r=-0.485) and total cholesterol (P=.043, r=-0.435) in hyperlipidemic subjects, which suggest that VK supplementation may have a positive effect in activating Gas6 protein and thereby reducing the aberrant plasma lipid levels. Further studies with high-fat diet (HFD)-fed animal model of hyperlipidemia demonstrated that VK supplementation (5 μg/kg body weight, 8 weeks) reduced the plasma lipid levels, stimulated both the plasma levels and the hepatic protein expression of Gla-Gas6 protein, and regulated the AMPK/SREBP1/PPARα signaling of hepatic lipid in HFD-fed mice. Moreover, by using (PA, 0.75 mM)-treated both control and GGCX knockdown hepatocytes, this study dissected the direct role of Gla-Gas6 in mediating the positive effect of VK on preventing the PA-induced impaired hepatic lipid via regulating AMPK/SREBP1/PPARα . Combining all, the present study demonstrated the beneficial effect of VK supplementation in preventing the impaired lipid homeostasis via activating VK-dependent Gas6 protein.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

is an intracellular signaling molecule involved in disease development.

Emerging evidence shows that (PA), a common fatty in the human diet, serves as a signaling molecule regulating the progression and development of many diseases at the molecular level. In this review, we focus on its regulatory roles in the development of five pathological conditions, namely, syndrome, cardiovascular diseases, cancer, neurodegenerative diseases, and inflammation. We summarize the clinical and epidemiological studies; and also the mechanistic studies which have identified the molecular targets for PA in these pathological conditions. Activation or inactivation of these molecular targets by PA controls disease development. Therefore, identifying the specific targets and signaling that are regulated by PA can give us a better understanding of how these diseases develop for the design of effective targeted therapeutics.

Keyword: metabolism

Encapsulation of soybean meal with fats enriched in and stearic acids: effects on rumen-undegraded protein and in vitro intestinal digestibility.

Fat coating of soybean meal (SBM) can reduce its protein degradability in the rumen, but the encapsulation of SBM with (PA) and stearic acids (SA) has not yet been investigated, despite both fatty acids are common energy sources in dairy cow diets. This study aimed to evaluate the effects of applying a novel method, using either 400 or 500\xa0g fat/kg (treatments FL40 and FL50, respectively), which was enriched in PA and SA at different ratios (100:0, 75:25, 50:50, 25:75 and 0:100), on physical and chemical characteristics, ruminal degradability, solubility and in vitro intestinal protein digestibility (IVIPD) of the obtained products. Encapsulation of SBM in fat resulted in greater mean particle size and lower bulk density and protein solubility than unprotected SBM (USBM). Treatment FL50 resulted in increased (p <\xa00.01) rumen-undegraded protein (RUP) compared to USBM. There were no differences in RUP of SBM when different PA: SA ratios were used. The mean RUP content of treatments FL40 and FL50 (306 and 349\xa0g/kg, respectively) was greater compared to USBM (262\xa0g/kg, p <\xa00.05), but lower than that for a standard heat-treated SBM (431\xa0g/kg). Values of IVIPD did not differ among SBM, heat-treated SBM and FL40 and FL50 samples, all being greater than 97.8%. In conclusion, encapsulation of SBM with fats enriched in PA and SA proved to be effective in reducing protein solubility and increasing RUP without depressing protein digestibility in the intestine. For validation of the method, in vivo research to investigate the effects of these products on the production of dairy cows is warranted.

Keyword: metabolism

Myo-inositol alters 13C-labeled fatty in human placental explants.

We postulate that myo-inositol, a proposed intervention for gestational-diabetes, affects transplacental lipid supply to the fetus. We investigated the effect of myo-inositol on fatty- processing in human placental-explants from uncomplicated pregnancies. Explants were incubated with 13C-labeled , 13C-oleic- and 13C-docosahexaenoic- across a range of myo-inositol concentrations for 24 h and 48 h. The incorporation of labeled-fatty-acids into individual lipids was quantified by liquid-chromatography-mass-spectrometry. At 24 h, myo-inositol increased the amount of 13C- and 13C-oleic- labeled lipids (median fold-change relative to control=1). Significant effects were seen with 30 µM myo-inositol (physiological) for 13C--lysophosphatidylcholines (1.26) and 13C--phosphatidylethanolamines (1.17). At 48 h, myo-inositol addition increased 13C-oleic--lipids but decreased 13C- and 13C-docosahexaenoic- lipids. Significant effects were seen with 30 µM myo-inositol for 13C-oleic--phosphatidylcholines (1.25), 13C-oleic--phosphatidylethanolamines (1.37) and 13C-oleic--triacylglycerols (1.32) and with 100 µM myo-inositol for 13C-docosahexaenoic--triacylglycerols (0.78). Lipids labeled with the same 13C-fatty- showed similar responses when tested at the same time-point, suggesting myo-inositol alters upstream processes such as fatty- uptake or activation. Myo-inositol supplementation may alter placental lipid physiology with unknown clinical consequences.

Keyword: metabolism

Polysaccharides Prevent -Evoked Apoptosis and Autophagy in Intestinal Porcine Epithelial Cell Line via Restoration of Mitochondrial Function and Regulation of MAPK and AMPK/Akt/mTOR Signaling Pathway.

polysaccharide (GLP) extracted from (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high-fat constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.

Keyword: metabolism

Exosomes derived from mangiferin‑stimulated perivascular adipose tissue ameliorate endothelial dysfunction.

Perivascular adipose tissue (PVAT) is considered to serve a vital role during the development of endothelial dysfunction. The current study investigated the effect of exosomes derived from mangiferin‑stimulated PVAT on endothelial function, including regeneration, migration, apoptosis and inflammation. The number of exosomes secreted by PVAT was increased by stimulation with mangiferin (0.1,\xa01\xa0or\xa010\xa0µM), and uptake of these exosomes by endothelial cells was observed. Exosomes produced by stimulation of PVAT with mangiferin reversed the effects of inflammation‑induced endothelial dysfunction following \xa0(PA) treatment. Furthermore, nuclear factor\xa0(NF)‑κB signaling in endothelial cells was significantly increased when treated with PA‑induced PVAT‑derived exosomes, whereas exosomes from the supernatant of PVAT stimulated with mangiferin reduced p65 and p50 phosphorylation levels in the cells, and inhibited p65 transportation to the nucleus. Taken together, the present study demonstrated that exosomes derived from mangiferin‑stimulated PVAT supernatant inhibited inflammation‑induced endothelial dysfunction via modulation of NF‑κB signaling.

Keyword: metabolism

Enhanced Fatty Scavenging and Glycerophospholipid Accompany Melanocyte Neoplasia Progression in Zebrafish.

Alterations in lipid in cancer cells impact cell structure, signaling, and energy , making lipid a potential diagnostic marker and therapeutic target. In this study, we combined PET, desorption electrospray ionization-mass spectrometry (DESI-MS), nonimaging MS, and transcriptomic analyses to interrogate changes in lipid in a transgenic zebrafish model of oncogenic RAS-driven melanocyte neoplasia progression. Exogenous fatty uptake was detected in melanoma tumor nodules by PET using the surrogate tracer 14(R,S)-18F-fluoro-6-thia-heptadecanoic ([18F]-FTHA), consistent with upregulation of genes associated with fatty uptake found through microarray analysis. DESI-MS imaging revealed that FTHA uptake in tumors was heterogeneous. Transcriptome and lipidome analyses further highlighted dysregulation of glycerophospholipid in melanoma tumor nodules, including increased abundance of phosphatidyl ethanolamine and phosphatidyl choline species, corroborated by DESI-MS, which again revealed heterogeneous phospholipid composition in tumors. Overexpression of the gene encoding lipoprotein lipase (LPL), which was upregulated in zebrafish melanocyte tumor nodules and expressed in the majority of human melanomas, accelerated progression of oncogenic RAS-driven melanocyte neoplasia in zebrafish. Depletion or antagonism of LPL suppressed human melanoma cell growth; this required simultaneous fatty synthase (FASN) inhibition when FASN expression was also elevated. Collectively, our findings implicate fatty acquisition as a possible therapeutic target in melanoma, and the methods we developed for monitoring fatty uptake have potential for diagnosis, patient stratification, and monitoring pharmacologic response. SIGNIFICANCE: These findings demonstrate the translational potential of monitoring fatty uptake and identify lipoprotein lipase as a potential therapeutic target in melanoma.©2019 American Association for Cancer Research.

Keyword: metabolism

Resveratrol Maintains Lipid Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver lipid disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver lipid disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive oxygen species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular lipid imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of lipid disorders relevant to the circadian clock.

Keyword: metabolism

[Atherosclerosis and atheromatosis are consequtive disordes. Pathology of the biological functions of trophology and endoecology is the basis for ischemic heart disease prevention.]

Atherosclerosis and atheromatosis are different nonphysiological processes with different etiology and pathogenesis. They manifest alterations in different biological functions. According to our original phylogenetic theory of general pathology, atherosclerosis is associated with altered biological function of trophology, eating, biological reaction of exotrophy. Atherosclerosis is induced by eating of nonoptimal for phylogenetically herbivorous Homo sapiens meat diet with high content of saturated fatty (SFA), which leads to in vivo formation of phylogenetically early low-efficient pathway of FA instead of highly-efficient oleic pathway operating in herbivores. Accumulation of nonligand very low density lipoproteins (VLDL) and low density lipoproteins (LDL) in the bloodstream results from nonphysiological reaction of compensation upon transport of SFA to cells. An increase in blood content of triglycerides (TG) and nonligand VLDLЛ→LDL coincides with the development of hypercholesterolemia: type IV→ type IIb → type V. Atheromatosis compensates changes in lipoproteins by activation of the biological function of endoecology (purity of the extracellular medium) in vivo, thus fulfilling the biological reaction of inflammation. This is physiological denaturation of apoВ-100 in nonligand VLDL→LDL by neutrophils via peroxidation, opsonization by the complement components, transcytosis across the endothelial monolayer and removal to the intima of elastic arteries that serves as a collection and utilization pool for phogogens from local intravascular pool of the intercellular medium. Endogenous phlogogens are utilized by phylogenetically early polyfunctional resident macrophages which are small in number and do not proliferate. Blood-borne monocytes-macrophages are also involved in this process, however, they do not express hydrolase of polyenic cholesteryl esters. Atheromatous masses are partially catabolized polyenic FA esterified by the alcohol cholesterol which were not internalized by cells. Atheromatosis is a process of pathological compensation in the realization of the function of endoecology. Prevention of atherosclerosis and atheromatosis should be based on elimination of the effects produced by a nonphysiological meat diet.

Keyword: metabolism

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy , but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on lipid , glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: metabolism

Vascular endothelial growth factor B inhibits lipid accumulation in C2C12 myotubes incubated with fatty acids.

To investigate (1) the effect of vascular endothelial growth factor B (VEGFB) on lipid accumulation and the alteration of fatty acids and fatty -related enzymes in C2C12 myotubes incubated with fatty acids and (2) the regulatory effect of VEGFB on skeletal muscle lipid . Mouse C2C12 myotubes were incubated with oleic (OA) and (PA), and differentiated mature C2C12 myotubes were treated with VEGFB. Oil-red O staining, BODIPY staining and cell triglycerides (TG) content were examined. Total RNA was isolated, and real-time PCR analysis was performed. Treatment with 100\u2009μM OA and 50\u2009μM PA induced lipid droplet accumulation and increased TG content (\u2009<\u2009.01), and 100\u2009ng/mL VEGFB reduced lipid droplet accumulation and decreased TG content (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of fatty transport protein 1 (FATP1) (\u2009<\u2009.01) and FATP4 (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of adipose TG lipase and hormone-sensitive lipase (\u2009<\u2009.01) as well as carnitine palmitoyltransferase I (\u2009<\u2009.01), peroxisome proliferator-activated receptor-γ coactivator-1α (\u2009<\u2009.01), acyl-coa dehydrogenase very long chain (\u2009<\u2009.05), acyl-coa synthetase long-chain family member 1 (\u2009<\u2009.01), peroxisomal acyl-coenzyme A oxidase 1 (\u2009<\u2009.05), and mitochondrial uncoupling protein 3 (\u2009<\u2009.01). VEGFB enhanced FATP1and FATP4 expression, promoted C2C12 myotube fatty oxidation and TG decomposition, and inhibited C2C12 myotube fatty re-esterification, thus inhibiting lipid accumulation in C2C12 myotubes incubated with fatty acids.

Keyword: metabolism

Effects of a new compound containing Palmitoylethanolamide and Baicalein in myocardial ischaemia/reperfusion injury in vivo.

Myocardial ischemia/reperfusion (I/R) injury is the principal cause of death, happens after prolonged obstruction of the coronary arteries. \xa0The first intervention to limit myocardial damage is directed to restoration of perfusion, to avoid inflammatory response and a significant oxidative stress triggered by infarction. Palmitoylethanolamide (PEA), is a well-known fatty amide-signaling molecule that possess an important anti-inflammatory and analgesic effects. PEA does not hold the ability to inhibit free radicals formation. Baicalein, a bioactive component isolated from a Chinese herbal medicine, has multiple pharmacological activities, such as a strong anti-oxidative effects.A combination of PEA and Baicalein could have beneficial effects on oxidative stress produced by inflammatory response.In the present study we explored the effects of composite containing PEA and Baicalein in a model of myocardial I/R injury.Myocardial ischemia/reperfusion injury was induced by occlusion of the left anterior descending coronary artery for 30\u202fmin followed by 2\u202fh of reperfusion. PEA-Baicalein (9:1), was administered (10\u202fmg/kg) 5\u202fmin before the end of ischemia and 1\u202fh after reperfusion.In this study, we clearly demonstrated that PEA-Baicalein treatment decreases myocardial tissue injury, neutrophils infiltration, markers for mast cell activation expression as chymase and tryptase and pro-inflammatory cytokines production (TNF-α, IL-1β). Moreover, PEA-Baicalein treatment reduces stress oxidative and modulates Nf-kB and apoptosis .These results support the idea that the association between PEA and Baicalein should be a potent candidate for the treatment of myocardial I/R injury.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: metabolism

Mitochondrial Reprogramming by CD36 Signaling Drives Macrophage Inflammatory Responses.

A hallmark of chronic inflammatory disorders is persistence of pro-inflammatory macrophages in diseased tissues. In atherosclerosis this is associated with dyslipidemia and oxidative stress, but mechanisms linking these phenomena to macrophage activation remain incompletely understood. To investigate mechanisms linking dyslipidemia, oxidative stress and macrophage activation through modulation of immunemetabolism, and to explore therapeutic potential targeting specific . Using a combination of biochemical, immunological, and ex vivo cell studies, we report that CD36 mediates a mitochondrial switch from oxidative phosphorylation to superoxide production in response to its ligand, oxLDL. Mitochondrial-specific inhibition of superoxide inhibited oxLDL-induced NF-κB activation and inflammatory cytokine generation. RNAseq, flow cytometry, H-labeled uptake, lipidomic analysis, confocal and EM imaging, and functional energetics revealed that oxLDL upregulated effectors of long-chain fatty (FA) uptake and mitochondrial import, while downregulating FA oxidation and inhibiting ATP5A, an electron transport chain (ETC) component. The combined effect is long-chain FA accumulation, alteration of mitochondrial structure and function, repurposing of the ETC to superoxide production, and NF-κB activation. Apoe null mice challenged with high fat diet showed similar changes in circulating Ly6C monocytes and peritoneal macrophages, along with increased CD36 expression. Moreover, mitochondrial ROS was positively correlated with CD36 expression in aortic lesional macrophages. These findings reveal that oxLDL/CD36 signaling in macrophages links dys-regulated FA to oxidative stress from the mitochondria, which drives chronic inflammation. Thus, targeting to CD36 and its downstream effectors may serve as potential new strategies against chronic inflammatory diseases such as atherosclerosis.

Keyword: metabolism

Interactions of fatty acids, nonsteroidal anti-inflammatory drugs, and coxibs with the catalytic and allosteric subunits of cyclooxygenases-1 and -2.

Prostaglandin endoperoxide H synthases-1 and -2, commonly called cyclooxygenases-1 and -2 (COX-1 and -2), catalyze the committed step in prostaglandin biosynthesis-the conversion of arachidonic to prostaglandin endoperoxide H Both COX isoforms are sequence homodimers that function as conformational heterodimers having allosteric (Eallo) and catalytic (Ecat) subunits. At least in the case of COX-2, the enzyme becomes folded into a stable Eallo/Ecat pair. Some COX inhibitors ( nonsteroidal anti-inflammatory drugs and coxibs) and common fatty acids (FAs) modulate Ecat activity by binding Eallo. However, the interactions and outcomes often differ between isoforms. For example, naproxen directly and completely inhibits COX-1 by binding Ecat but indirectly and incompletely inhibits COX-2 by binding Eallo. Additionally, COX-1 is allosterically inhibited up to 50% by common FAs like , whereas COX-2 is allosterically activated 2-fold by . FA binding to Eallo also affects responses to COX inhibitors. Thus, COXs are physiologically and pharmacologically regulated by the FA tone of the milieu in which each operates-COX-1 in the endoplasmic reticulum and COX-2 in the Golgi apparatus. Cross-talk between Eallo and Ecat involves a loop in Eallo immediately downstream of Arg-120. Mutational studies suggest that allosteric modulation requires a direct interaction between the carboxyl group of allosteric effectors and Arg-120 of Eallo; however, structural studies show some allosterically active FAs positioned in COX-2 in a conformation lacking an interaction with Arg-120. Thus, many details about the biological consequences of COX allosterism and how ligand binding to Eallo modulates Ecat remain to be resolved.© 2019 Smith and Malkowski.

Keyword: metabolism

Metabolomics reveals potential biomarkers in the rumen fluid response to different milk production of dairy cows.

In the present study, an LC/MS metabolomics approach was performed to investigate potential biomarkers of milk production in high- and low-milk-yield dairy cows and to establish correlations among rumen fluid metabolites; the results of this study provide insights into the mechanisms underlying the milk production-related characteristics of rumen fluid in dairy cows. Sixteen lactating dairy cows with similar parity and days in milk were divided into high-yield (HY) and low-yield (LY) groups based on milk yield. On day 21, rumen fluid metabolites were quantified applying LC/MS. The principal component analysis (PCA) and orthogonal partial least-squares (OPLS-DA) showed significantly separated clusters of the ruminal metabolite profiles of HY and LY groups. Compared with HY group, a total of 24 ruminal metabolites were significantly greater in LY group, such as 3-hydroxyanthranilic , carboxylic acids, carboxylic derivatives (L-isoleucine, L-valine, L-tyrosine, etc.), diazines (uracil, thymine, cytosine), and , while the concentrations of 30 metabolites were dramatically decreased in LY group compared to HY group, included gentisic , caprylic , and myristic . The metabolite enrichment analysis indicated that protein digestion and absorption, ABC transporters and unsaturated fatty biosynthesis were significantly different between the two groups. Correlation analysis between the ruminal microbiome and metabolites revealed that certain typical metabolites were exceedingly associated with definite ruminal bacteria; Firmicutes, Actinobacteria and Synergistetes phyla were highly correlated with most metabolites. These findings revealed that the ruminal metabolite profiles were significantly different between HY and LY groups, and these results may provide novel insights to evaluate biomarkers for a better feed digestion and may reveal the potential mechanism underlying the difference in milk yield in dairy cows.

Keyword: metabolism

Serum C16:1n7/C16:0 ratio as a diagnostic marker for non-alcoholic steatohepatitis.

Accurate diagnosis of non-alcoholic steatohepatitis (NASH) from non-alcoholic fatty liver disease (NAFLD) is clinically important. Therefore, there is a need for easier ways of diagnosing NASH. In this study, we investigated the serum fatty composition and evaluated the possibility of using the serum fatty composition as a diagnostic marker of NASH.The subjects were 78 NAFLD patients (non-alcoholic fatty liver [NAFL]: 30, NASH: 48) and 24 healthy individuals. Fatty acids extracted from the liver tissue and serum were identified and quantified by gas chromatography. In addition, we evaluated the relationship between serum and liver tissue fatty composition, patient background, and liver histology. The diagnostic performance of NASH was evaluated by calculating the area under the receiver operating characteristic (AUROC).The results of the fatty analysis showed the C16:1n7/C16:0 ratio to have the strongest correlation between serum and liver tissue (r\xa0=\xa00.865, P\xa0<\xa00.0001). The serum C16:1n7/C16:0 ratio in the NASH group was higher compared with that in the NAFL group (P\xa0=\xa00.0007). Evaluation of the association of the serum C16:1n7/C16:0 ratio with liver histology revealed significant correlation with lobular inflammation score, ballooning score, and fibrosis score. The AUROC for predicting NASH in all NAFLD patients was 0.7097. The AUROC was nearly equivalent even when the study subjects were restricted to patients with a fibrosis score\xa0≤\xa02 only (AUROC 0.6917).Measuring the serum C16:1n7/C16:0 ratio may be an effective non-invasive method for diagnosing NASH, particularly in its early stages.© 2019 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Keyword: metabolism

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular Inflammation in HepG2 Cells.

Fatty acids induced hepatic inflammation plays an important role in nonalcoholic fatty liver disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti-inflammation in various human organs. However, the anti-inflammation property of HS in the fatty liver is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free fatty acids-induced inflammation. The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: metabolism

Uncovering sperm metabolome to discover biomarkers for bull fertility.

Subfertility decreases the efficiency of the cattle industry because artificial insemination employs spermatozoa from a single bull to inseminate thousands of cows. Variation in bull fertility has been demonstrated even among those animals exhibiting normal sperm numbers, motility, and morphology. Despite advances in research, molecular and cellular mechanisms underlying the causes of low fertility in some bulls have not been fully elucidated. In this study, we investigated the profile of bull spermatozoa using non-targeted metabolomics. Statistical analysis and bioinformatic tools were employed to evaluate the profiles high and low fertility groups. associated with the sperm metabolome were also reported.A total of 22 distinct metabolites were detected in spermatozoa from bulls with high fertility (HF) or low fertility (LF) phenotype. The major metabolite classes of bovine sperm were organic acids/derivatives and fatty acids/conjugates. We demonstrated that the abundance ratios of five sperm metabolites were statistically different between HF and LF groups including gamma-aminobutyric (GABA), carbamate, benzoic , lactic , and . Metabolites with different abundances in HF and LF bulls had also VIP scores of greater than 1.5 and AUC- ROC curves of more than 80%. In addition, four associated with differential metabolites namely alanine, aspartate and glutamate , β-alanine , glycolysis or gluconeogenesis, and pyruvate were also explored.This is the first study aimed at ascertaining the metabolome of spermatozoa from bulls with different fertility phenotype using gas chromatography-mass spectrometry. We identified five metabolites in the two groups of sires and such molecules can be used, in the future, as key indicators of bull fertility.

Keyword: metabolism

Chronic intake of moderate fat-enriched diet induces fatty liver and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of disorders, such as overweight/obesity and syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: metabolism

Lipid production by Lipomyces starkeyi using sap squeezed from felled old oil palm trunks.

The ability of oleaginous yeast Lipomyces starkeyi to efficiently produce lipids when cultivated on sap extracted from felled oil palm trunk (OPT) as a novel inexpensive renewable carbon source was evaluated. OPT sap was found to contain approximately 98\xa0g/L glucose and 32\xa0g/L fructose. Batch fermentations were performed using three different OPT sap medium conditions: regular sap, enriched sap, and enriched sap at pH 5.0. Under all sap medium conditions, the cell biomass and lipid production achieved were approximately 30\xa0g/L and 60% (w/w), respectively. L.\xa0starkeyi tolerated acidified medium (initial pH ≈ 3) and produced considerable amounts of ethanol as well as xylitol as by-products. The fatty profile of L.\xa0starkeyi was remarkably similar to that of palm oil, one of the most common vegetable oil feedstock used in biodiesel production with oleic as the major fatty followed by , stearic and linoleic acids.Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Keyword: metabolism

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on lipid and inflammatory signaling in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: metabolism

Fatty Profile and Cardiometabolic Markers in Relation with Diet Type and Omega-3 Supplementation in Spanish Vegetarians.

Plant-based diets are becoming increasingly popular, and scientific information concerning the nutritional status in this population is needed. This study determined the fatty profile of Spanish lacto-ovo vegetarians (LO-vegetarians) and vegans. Participants were 104 healthy adults, LO-vegetarians ( = 49) and vegans ( = 55). Lifestyle habits and consumption of food and omega-3 supplements were estimated by questionnaires. BMI, blood pressure, and abdominal and body fat were determined. Serum was collected to analyze fatty acids, glucose, lipids, homocysteine, insulin, and leptin. Volunteers were classified according to serum omega-6 to omega-3 (-6/-3) ratio into three groups: -6/-3 < 10, -6/-3 ≥ 10 to 20, and -6/-3 > 20. Results showed low cardiovascular risk and high insulin sensitivity with negligible differences between diet types. Linoleic (C18:2-6) was the major serum fatty , followed by oleic (C18:1-9) and (C16:0) acids. In contrast, serum eicosapentaenoic (EPA, C20:5-3) and docosahexaenoic (DHA, C22:6-3) were (median, interquartile range) 0.27, 0.18% and 1.59, and 0.93%, respectively. Users of -3 supplements (<10% of total vegetarians) had significantly higher EPA than non-users, while frequent consumption of flax-seeds was associated with increased α-linolenic (C18:3-3). However, neither -3 supplementation nor food consumption affected DHA levels in this vegetarian population.

Keyword: metabolism

Berberine attenuates sodium palmitate-induced lipid accumulation, oxidative stress and apoptosis in grass carp(Ctenopharyngodon idella)hepatocyte in vitro.

The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, lipid accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in lipid accumulation-hepatocytes induced by sodium palmitate in vitro. The lipid accumulation-hepatocytes (induced by 0.5\u202fmM sodium palmitate for 24\u202fh) were treated with 5\u202fμM berberine for 12\u202fh. Then, the Cell viability, intracellular triglyceride (TG) content, lipid peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, lipid droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from mitochondria into the cytoplasm. Beberine could improve intracellular lipid droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.Copyright © 2019. Published by Elsevier Ltd.

Keyword: metabolism

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 diabetes. Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and lipid . Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: metabolism

Enzymatic preparation of structured triacylglycerols with arachidonic and acids at the sn-2 position for infant formula use.

Structured TAGs with and polyunsaturated fatty (PUFA) at the sn-2 position have various health benefits for infants. In this study, we first compared two enzymatic routes for preparation of the structured TAGs. Results showed that the one-pot and two-step syntheses led to 37.6% and 55.4% oleic incorporation, respectively, after 10\u202fh and reaction route had little effect on the sn-2 fatty composition. Subsequently, reaction variables of the two-step synthesis were optimized. Under the optimal conditions, 53.5% oleic was incorporated into the structured TAGs after 6-h acidolysis. Major fatty acids at the sn-2 position were (68.7%), ARA (9.8%) and oleic (7.9%). This is the first study reporting a two-step enzymatic method for structured TAGs preparation. Compared to the one-pot synthesis, current method significantly improves the efficiency of the acidolysis by product inhibition elimination. The synthetic TAGs have potential use in infant formulas.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: metabolism

MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue inflammation in obesity. Both MIP-1α and free fatty acids are elevated in obesity/T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling . In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in obesity/T2D may contribute to inflammation through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: metabolism

Glycerolipid Composition of the Red Macroalga and Comparison to the Closely Related Producing Different Types of Eicosanoids.

The red macroalga is a well-known producer of eicosanoids such as hydroxyeicosatetraenoic acids, but the alga produces almost no prostaglandins, unlike the closely related . This indicates that the related two algae would have different enzyme systems or substrate composition. To carry out more in-depth discussions on the pathway of eicosanoids between the two algae, we investigated the characteristics of glycerolipids, which are the substrates of eicosanoids production, of and compared them to the reported values of . In , monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylcholine (PC) were the major lipid classes and accounted for 44.4% of the total lipid extract. The predominant fatty acids were arachidonic (20:4n-6), an eicosanoids precursor, and (16:0). The 20:4n-6 content was extremely high in MGDG and PC (>70%), and the 16:0 content was extremely high in DGDG and SQDG (>40%). A chiral-phase HPLC analysis showed that fatty acids were esterified at the -1 and -2 positions of those lipids. The glycerolipid molecular species were determined by reversed-phase HPLC⁻ESI⁻MS analysis. The main glycerolipid molecular species were 20:4n-6/20:4n-6 (-1/-2) for MGDG (63.8%) and PC (48.2%), 20:4n-6/16:0 for DGDG (71.1%) and SQDG (29.4%). These lipid characteristics of were almost the same as those of . Hence, the differences of the eicosanoids producing ability between the two algae would not be due to the difference of substrate composition but the difference of enzyme system.

Keyword: metabolism

Serum metabolome and targeted bile profiling reveals potential novel biomarkers for drug-induced liver injury.

This study aims to determine the non-invasive, reliable and sensitive biochemical parameters for the diagnosis of drug-induced liver injury (DILI).Ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) and selected reaction monitoring (SRM) were used to profile the serum metabolome and quantify 15 targeted bile metabolites, respectively, in samples obtained from 38 DILI patients and 30 healthy controls.A comparison of the resulting serum metabolome profiles of the study participants revealed significant differences between DILI patients and healthy controls. Specifically, serum , taurochenodeoxycholic , glycocholic (GCA), and tauroursodeoxycholic (TUDCA) levels were significantly higher, and serum lysophosphatidylethanolamine levels were significantly lower in DILI patients vs healthy controls (P\u200a<\u200a.001). Furthermore, the SRM assay of bile acids revealed that the increase in GCA, taurocholic (TCA), TUDCA, glycochenodeoxycholic (GCDCA), glycochenodeoxycholic sulfate (GCDCS), and taurodeoxycholic (TDCA) corresponded to a higher degree of liver damage. These results also indicate that serum concentrations of chenodeoxycholic (CDCA), deoxycholic (DCA) and lithocholic (LCA) were significantly lower in patients with severe DILI, when compared to healthy controls, and that this decrease was closely correlated to the severity of liver damage.Taken together, these results demonstrate that bile acids could serve as potential biomarkers for the early diagnosis and severity of DILI.

Keyword: metabolism

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of obesity, insulin resistance, type-2 diabetes and syndrome. Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: metabolism

Electric pulse stimulation inhibited lipid accumulation on C2C12 myotubes incubated with oleic and .

To investigate the effect of electrical pulse stimulation (EPS) on lipid accumulation and alteration of fatty -related enzymes in C2C12 myotubes incubated with fatty acids. Mouse C2C12 myotubes were incubated with oleic and , and differentiated C2C12 myotubes were treated with EPS, oil-red O (ORO), BODIPY staining and triglyceride (TG) content were examined. Total RNA was isolated, and real-time polymerase chain reaction analysis was performed. (1) EPS decreased TG content (\u2009<\u2009.01). (2) EPS significantly induced the mRNA expression of FAD/CD36 (\u2009<\u2009.05), FATP4 (\u2009<\u2009.001), FABP1 (\u2009<\u2009.01) and FABP5 (\u2009<\u2009.01). (3) EPS significantly inhibited the mRNA expression of fatty synthase (\u2009<\u2009.01). (4) Adipose triglyceride lipase and hormone-sensitive lipase expression were significantly elevated (\u2009<\u2009.001), and induced the mRNA expression of CPT1 (\u2009<\u2009.01), ACOX1 (\u2009<\u2009.05), UCP3 (\u2009<\u2009.05) and PPARα (\u2009<\u2009.001) after EPS. EPS reduced lipid droplet accumulation; enhanced CD36, FATP4, FABP1 and FABP5 expression; inhibited C2C12 myotube fatty re-esterification; and promoted fatty oxidation in C2C12 myotubes.

Keyword: metabolism

Evaluation of anti-nociceptive and anti-inflammatory activities of the methanol extract of Holigarna caustica (Dennst.) Oken leaves.

Holigarna caustica (Dennst.) is commonly used in traditional medicine to treat a variety of painful conditions such as eye irritation, inflammation, arthritis, skin diseases, cuts and wounds.The present study was undertaken to investigate the anti-nociceptive and anti-inflammatory activities of the methanol extract of H. caustica leaves and to elucidate its possible mechanism(s) of action.Fresh leaves of H. caustica were collected, dried, and extracted with methanol (MEHC). MEHC was subjected to activity testing, using chemical-induced (acetic and formalin test) and heat-induced (hot plate and tail immersion test) pain models. To determine the possible mechanism behind the anti-nociceptive activity of MEHC, the opioid antagonist naltrexone was used to evaluate the involvement of opioid receptors in the case of formalin, hot plate and tail immersion tests, while the involvement of the cGMP and ATP-sensitive K channel were assessed using methylene blue and glibenclamide respectively, in the acetic -induced writhing test. In parallel, the carrageenan-induced paw oedema model was used to determine the anti-inflammatory potential of the extract. Exploratory and motor behaviours were evaluated by the open-field test. Various bioactive compounds potentially responsible for the anti-nociceptive and anti-inflammatory activities were ascertained using GC-MS analysis.MEHC showed strong, significant and dose-dependent anti-nociceptive activity in all chemical-induced and heat-induced pain models at all experimental doses. The association of opioid receptors with the observed anti-nociceptive effects was confirmed by using naltrexone. The cGMP and ATP-sensitive K channel pathway was also shown to be involved in the anti-nociceptive activity of MEHC. In addition, MEHC exhibited a dose-dependent inhibition of inflammatory oedema induced by carrageenan. MEHC was not connected with changes in either the locomotor activity or motor responses of mice. In a GC-MS analysis, 40 compounds were identified, among which twelve are documented bioactive compounds with potent analgesic and anti-inflammatory properties.Our current study revealed that MEHC possesses strong central and peripheral anti-nociceptive as well as anti-inflammatory activity. It may also be concluded that both opioid receptors as well as the cGMP and ATP-sensitive K channel pathway are involved in the anti-nociceptive mechanism of MEHC. This study rationalizes the ethnomedicinal use of H. caustica leaves in various painful conditions.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: metabolism

Optimization of Synechococcus sp. VDW Cultivation with Artificially Prepared Shrimp Wastewater for Ammonium Removal and Its Potential for Use As a Biofuel Feedstock.

To investigate the potential of application of marine cyanobacterium for concurrent biomass production and ammonium removal, Synechococcus sp. VDW was cultured under different conditions in medium containing varying concentrations of NHCl. Response surface methodology (RSM) was then used to build a predictive model of the combined effects of independent variables (pH, inoculum size, ammonium concentration). At the optimum conditions of initial pH 7.4, inoculum size 0.17 (OD730) and ammonium concentration 10.5 mg L, the maximum ammonium removal and biomass productivity were about 95% and 34 mg Ld, respectively, after seven days of cultivation. The result of fatty methyl ester (FAME) analysis showed that the major fatty acids were (C16:0), linoleic (C18:2 n6 cis), palmitoleic (C16:1) and oleic (C18:1 n9 cis), which accounted for more than 80% weight of total fatty acids. Further, analysis of neutral lipid accumulation using flow cytometry revealed that the mean of the fluorescence intensity increased under optimal conditions. These results indicate that Synechococcus sp. VDW has the potential for use for concurrent water treatment and production of biomass that can be applied as biofuel feedstock.

Keyword: metabolism

Potential of fungal metabolites as a biocontrol agent against cotton aphid, Aphis gossypii Glover and the possible mechanisms of action.

The present study investigated the insecticidal activity of the different organic extracts from the entomopathogenic fungi, Cladosporium cladosporioides, Metarhizium anisopliae, Purpureocillium lilacinum, and Trichoderma longibrachiatum towards cotton aphid, Aphis gossypii. The methanol extracts from the mycelia and spores of C. cladosporioides and P. lilacinum exhibited the highest insecticidal activity against A. gossypii compared with other extracts, which LC values were recorded to be 57.60 and 94.18\u202fppm, respectively. The major constituents identified in both methanol extracts by GC-MS analysis were linoleic and . The methanol extracts of C. cladosporioides and P. lilacinum caused a voluminous increase in the total carbohydrates content of A. gossypii adults, while the total protein content was significantly decreased by both extracts. The activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were significantly reduced by methanol extracts. The P. lilacinum extract caused a considerable reduction in the activity of glutathione-S-transferase (GST), α- and β-esterase by 28.9, 27.9 and 23.4%, respectively. Both extracts induced a significant increase in phenoloxidase and chitinase activity of A. gossypii adults. These results suggest that C. cladosporioides and P. lilacinum methanol extracts could be used as a promising approach for the management of A. gossypii in many economically crops.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid .LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: metabolism

Effects of Chlorpyrifos on Cholinesterase and Serine Lipase Activities and Lipid in Brains of Rainbow Trout (Oncorhynchus mykiss).

Chlorpyrifos is an organophosphorus insecticide that elicits acute toxicity through inhibition of acetylcholinesterase (AChE), leading to acetylcholine accumulation and prolonged stimulation of cholinergic receptors throughout the central and peripheral nervous systems. Previous studies have indicated that neurodevelopment may also be impaired through alternative , including reduction of cAMP catalyzed downstream events. The upstream initiating events that underlie non-cholinergic neurological actions of chlorpyrifos and other organophosphorus compounds remain unclear. To investigate the potential role of disruption of fatty signaling as a mechanism of toxicity, lipid and fatty profiles were examined to identify alterations that may play a critical role in upstream signaling in the CNS. Juvenile rainbow trout were treated for 7 days with nominal chlorpyrifos concentrations previously reported to diminish olfactory responses (10, 20, and 40\u2009μg/L). While lethality was noted higher doses, measured chlorpyrifos concentrations of 1.38\u2009μg/L (nominal concentration 10\u2009μg/L) significantly reduced the activity of AChE and two serine lipases, monoacylglycerol lipase and fatty amide hydrolase in the brain. Reductions in lysophosphatidylethanolamines (16:0; 18:0, 18:1, and 22:6) derived from the phosphatidylethanolamines and free fatty acids ( acid16:0; Linolenic acid18:3; Eicosadienoic 20:2; Arachidonic 20:4; and Docosahexaenoic 22:6) were also noted, suggesting that chlorpyrifos inhibited the of selected phospholipid signaling precursors at sublethal concentrations. These results indicate that in addition to AChE inhibition, environmentally relevant chlorpyrifos exposure alters serine lipase activity and lipid metabolites in the trout brain, which may compromise neuronal signaling and impact neurobehavioral responses in aquatic animals.© The Author(s) 2019. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Keyword: metabolism

Hexadecenoic Fatty Positional Isomers and De Novo PUFA Synthesis in Colon Cancer Cells.

involves delta-9 and delta-6 desaturase enzymes forming palmitoleic (9-16:1; -7 series) and sapienic (6-16:1; -10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty (MUFA) isomers are under development. Furthermore, sapienic can bring to the de novo synthesis of the -10 polyunsaturated fatty (PUFA) sebaleic (5,8-18:2), but such transformations in cancer cells are not known. The model of Caco-2 cell line was used to monitor sapienic supplementation (150 and 300 μM) and provide evidence of the formation of -10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and acids evidenced that lipid remodelling was influenced by the type of fatty and positional isomer, with an increase of 8-18:1, -10 PUFA and a decrease of saturated fats in case of sapienic . Cholesteryl esters were formed only in cases with sapienic . Sapienic was the less toxic among the tested fatty acids, showing the highest ECs and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic increases the distribution of fluid regions, probably connected with the formation of 8-18:1 and the -10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and cancer research.

Keyword: metabolism

CHP1 Regulates Compartmentalized Glycerolipid Synthesis by Activating GPAT4.

Cells require a constant supply of fatty acids to survive and proliferate. Fatty acids incorporate into membrane and storage glycerolipids through a series of endoplasmic reticulum (ER) enzymes, but how these enzymes are regulated is not well understood. Here, using a combination of CRISPR-based genetic screens and unbiased lipidomics, we identified calcineurin B homologous protein 1 (CHP1) as a major regulator of ER glycerolipid synthesis. Loss of\xa0CHP1 severely reduces fatty incorporation and storage in mammalian cells and invertebrates. Mechanistically, CHP1 binds and activates GPAT4, which catalyzes the initial rate-limiting step in glycerolipid synthesis. GPAT4 activity requires CHP1 to be N-myristoylated, forming a key molecular interface between the two proteins. Interestingly, upon CHP1 loss, the peroxisomal enzyme, GNPAT, partially compensates for the loss of ER lipid synthesis, enabling cell proliferation. Thus, our work identifies a conserved regulator of glycerolipid and reveals plasticity in lipid synthesis of proliferating cells.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: metabolism

Berberine Protects Glomerular Podocytes via Inhibiting Drp1-Mediated Mitochondrial Fission and Dysfunction.

Elevated levels of plasma free fatty (FFA) and disturbed mitochondrial dynamics play crucial roles in the pathogenesis of diabetic kidney disease (DKD). However, the mechanisms by which FFA leads to mitochondrial damage in glomerular podocytes of DKD and the effects of Berberine (BBR) on podocytes are not fully understood. : Using the db/db diabetic mice model and cultured mouse podocytes, we investigated the molecular mechanism of FFA-induced disturbance of mitochondrial dynamics in podocytes and testified the effects of BBR on regulating mitochondrial dysfunction, podocyte apoptosis and glomerulopathy in the progression of DKD. : Intragastric administration of BBR for 8 weeks in db/db mice significantly reversed glucose and lipid disorders, podocyte damage, basement membrane thickening, mesangial expansion and glomerulosclerosis. BBR strongly inhibited podocyte apoptosis, increased reactive oxygen species (ROS) generation, mitochondrial fragmentation and dysfunction both and . Mechanistically, BBR could stabilize mitochondrial morphology in podocytes via abolishing (PA)-induced activation of dynamin-related protein 1 (Drp1). : Our study demonstrated for the first time that BBR may have a previously unrecognized role in protecting glomerulus and podocytes via positively regulating Drp1-mediated mitochondrial dynamics. It might serve as a novel therapeutic drug for the treatment of DKD.

Keyword: metabolism

The association of adelmidrol with sodium hyaluronate displays beneficial properties against bladder changes following spinal cord injury in mice.

The disruption of coordinated control between the brain, spinal cord and peripheral nervous system caused by spinal cord injury (SCI) leads to several secondary pathological conditions, including lower urinary tract dysfunction. In fact, urinary tract dysfunction associated with SCI is urinary dysfunction could be a consequence of a lack of neuroregeneration of supraspinal that control bladder function. The object of the current research was to explore the effects of adelmidrol + sodium hyaluronate, on bladder damage generated after SCI in mice. Spinal cord was exposed via laminectomy, and SCI was induced by extradural compression at T6 to T7 level, by an aneurysm clip with a closing force of 24 g. Mice were treated intravesically with adelmidrol + sodium hyaluronate daily for 48 h and 7 days after SCI. Adelmidrol + sodium hyaluronate reduced significantly mast cell degranulation and down-regulated the nuclear factor-κB pathway in the bladder after SCI both at 48 h and 7days. Moreover, adelmidrol + sodium hyaluronate reduced nerve growth factor expression, suggesting an association between neurotrophins and bladder pressure. At 7 days after SCI, the bladder was characterized by a marked bacterial infection and proteinuria; surprisingly, adelmidrol + sodium hyaluronate reduced significantly both parameters. These data show the protective roles of adelmidrol + sodium hyaluronate on bladder following SCI, highlighting a potential therapeutic target for the reduction of bladder changes.

Keyword: metabolism

-Rich High Fat Diet Exacerbates Experimental Pulmonary Fibrosis by Modulating Endoplasmic Reticulum Stress.

The impact of lipotoxicity on the development of lung fibrosis is unclear. Saturated fatty acids such as (PA) activate endoplasmic reticulum (ER) stress, a cellular stress response associated with the development of idiopathic pulmonary fibrosis (IPF). We tested the hypothesis that PA increases susceptibility to lung epithelial cell death and experimental fibrosis by modulating ER stress. Total liquid chromatography and mass spectrometry were used to measure fatty content in IPF lungs. Wild-type mice were fed a high fat diet (HFD) rich in PA or a standard diet (SD) and subjected to bleomycin-induced lung injury. Lung fibrosis was determined by hydroxyproline content. Mouse lung epithelial cells were treated with PA. ER stress and cell death were assessed by Western blotting, TUNEL staining and cell viability assays. IPF lungs had a higher level of PA compared to controls. Bleomycin-exposed mice fed a HFD had significantly increased pulmonary fibrosis associated with increased cell death and ER stress compared to those fed a SD. PA increased apoptosis and activation of the unfolded protein response in lung epithelial cells. This was attenuated by genetic deletion and chemical inhibition of CD36, a fatty transporter. In conclusion, consumption of a HFD rich in saturated fat increases susceptibility to lung fibrosis and ER stress and PA mediates lung epithelial cell death and ER stress via CD36. These findings demonstrate that lipotoxicity may have a significant impact on the development of lung injury and fibrosis by enhancing pro-death ER stress .

Keyword: metabolism

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

Keyword: metabolism

Fatty levels alterations in THP-1 macrophages cultured with lead (Pb).

As cardiovascular events are one of the main causes of death in developed countries, each factor potentially increasing the risk of cardiovascular disease deserves special attention. One such factor is the potentially atherogenic effect of lead (Pb) on lipid , and is significant in view of the still considerable Pb environmental pollution and the non-degradability of Pb compounds.Analysis of saturated fatty acids (SFA) (caprylic (C8:0), decanoic (C10:0), lauric (C12:0), tridecanoic (C13:0), myristic (C14:0), pentadecanoic (C15:0), (C16:0), heptadecanoic (C17:0), stearic (C18:0), and behenic (C22:0)), monounsaturated fatty (MUFA) (palmitoleic (C16:1), oleic (18:1w9), trans-vaccenic (C18:1 trans11)), and polyunsaturated fatty (PUFA) (linoleic (C18:2n6), gamma-linolenic (C18:3n6), arachidonic (C20:4n6)), was conducted by gas chromatography. Analysis of stearoyl-CoA desaturase (SCD), fatty desaturase 1 (FADS1) and fatty desaturase 2 (FADS2) expression was performed using qRT-PCR. Oxidative stress intensity (malondialdehyde - MDA concentration) was measured using spectrophotometric method. Intracellular generation of reactive oxygen species (ROS) in macrophages was visualized by fluorescence microscopy and quantitatively measured by plate reader.Pb caused quantitative alterations in FAs profile in macrophages; the effect was Pb-concentration dependent and selective (i.e. concerned only selected FAs). In general, the effect of Pb was biphasic, with Pb levels of 1.25\u2009μg/dL and 2.5\u2009μg/dL being stimulatory, and 10\u2009μg/dL being inhibitory on concentrations of selected FAs. The most potent Pb concentration, resulting in increase in levels of 9 FAs, was 2.5\u2009μg/dL, the Pb-level corresponding to the mean blood Pb concentrations of people living in urban areas not contaminated by Pb. Pb was found to exert similar, biphasic effect on the expression of FADS1. However, Pb decreased, in a concentration-dependent manner, the expression of SCD and FADS2. Pb significantly increased MDA and ROS concentration in macrophages.Environmental Pb exposure might be a risk factor resulting in alterations in FAs levels, oxidative stress and increased MDA concentration in macrophages, which might lead to the formation of foam cells and to inflammatory reactions.Copyright © 2019 Elsevier GmbH. All rights reserved.

Keyword: metabolism

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving lipid , antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis (NASH) is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high fat high fructose (HFHF) diet fed C57BL/6J mice, used as model of NASH. The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved lipid homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, mitochondria are intracellular source of reactive oxygen species (ROS) and important sites of lipid , we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti-NASH potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: metabolism

Mutation of IDH1 aggravates the fatty ‑induced oxidative stress in HCT116 cells by affecting the mitochondrial respiratory chain.

Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the reprogramming of cancer cells; however their functions in lipid remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic (OA) or (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive oxygen species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of mitochondria, immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty ‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.

Keyword: metabolism

Apigenin reduces the excessive accumulation of lipids induced by via the AMPK signaling pathway in HepG2 cells.

In recent years, increasing attention has been paid to diseases caused by excessive accumulation of lipids in the liver with therapeutic agents derived from natural products offering an alternative treatment to conventional therapies. Among these therapeutic agents, apigenin, a natural flavonoid, has been proven to exert various beneficial biological effects. In the present study, the antiadipogenic effects of apigenin in HepG2 cells was investigated. It was demonstrated that the treatment of cells with different concentrations of apigenin for 24 h significantly decreased the -induced increases in total cholesterol (TC) and triglyceride (TG) levels as well as intracellular lipid accumulation. In addition, apigenin increased the phosphorylated-AMP-activated protein kinase (AMPK) levels but decreased the expression levels of 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding protein (SREBP)-1, fatty synthase, and SREBP-2 in a concentration-dependent manner. The present findings suggested that apigenin might improve lipid by activating the AMPK/SREBP pathway to reduce lipid accumulation in the liver.Copyright: © Lu et al.

Keyword: metabolism

The effect of saturated and unsaturated fatty acids on the production of outer membrane vesicles from and .

The aim of present study is to investigate the effect of fatty acids on the outer membrane vesicles (OMVs) produced by spp. spp. is the important member of Gut that employ OMVs production for interact with host. Besides, dietary fatty acids could influence on determination of gut composition and immune response. In this regard, we evaluated the effect of fatty acids on the growth and OMVs production of and . and were grown on BHI broth with and without and palmitoleic acids as saturated and unsaturated fatty acids, respectively. OMVs were extracted using multiple centrifugation and tris-ethylene diamine tetra acetic (EDTA)-Sodium deoxy cholate buffers. Physicochemical properties of OMVs were detected by electron microscopy (SEM), Bradford Coomassie brilliant blue assay and SDS-PAGE. Data were analyzed with One-way ANOVA using SPSS.The growths of both were significantly increased by . Nevertheless, palmitoleic had no significant effect on them. significantly decreased and increased the production of OMVs at low and high concentration, respectively. However, the production of OMVs was not significantly affected by . Although palmitoleic had a significant decreasing effect on the production of OMVs, it significantly increased the production of OMVs at low concentration.In conclusion we reported that had a stimulatory effect on the growth of and and had a dose dependent effect on the production of OMVs. Also producing of OMVs was affected by palmitoleic in a dose dependent manner.

Keyword: microbiome

The impact of dietary sn-2 triacylglycerols in combination with docosahexaenoic or arachidonic on lipid metabolism and host faecal composition in Sprague Dawley rats.

Sn-2 triacylglycerols (sn2PA fat) and polyunsaturated fatty acids are thought to influence the metabolic status and intestinal bacterial population of the host. In this study, the impact of sn2PA fat in combination with DHA or ARA in the diet on lipid metabolism in the liver and faecal composition were investigated in rats fed diets containing sn2PA fat, 90% sn2PA fat + 10% DHA oil (wt%), or 90% sn2PA fat + 10% ARA oil (wt%). Tissue fatty composition was measured using gas chromatography (GC), whereas the faecal microbial composition was assessed using 16S rRNA high-throughput sequencing technology. In addition, faecal short-chain fatty acids (SCFA) were analyzed using ion chromatography. The results showed that sn2PA fat in combination with DHA or ARA significantly reduced liver triacylglyceride (TG) content compared with the sn2PA fat only group. Moreover, the supplementation with sn2PA fat in combination with DHA or ARA significantly promoted the growth of Lactobacillus in the feces at the genus level. On the other hand, the growth of the opportunistic pathogen Desulfovibrio was significantly inhibited by sn2PA fat in combination with ARA compared with the sn2PA fat group. In addition, sn2PA fat in combination with DHA or ARA significantly increased total SCFA concentration in the faeces, suggesting a beneficial effect on host intestinal health.

Keyword: microbiome

Yeast Smell Like What They Eat: Analysis of Volatile Organic Compounds of in Growth Media Supplemented with Different Lipids.

is part of the human skin . Its volatile organic compounds (VOCs) possibly contribute to the characteristic odour in humans, as well as to interaction. The aim of this study was to investigate how the lipid composition of the liquid medium influences the production of VOCs. Growth was performed in four media: (1) mDixon, (2) oleic (OA), (3) oleic + (OA+PA), and (4) (PA). The profiles of the VOCs were characterized by HS-SPME/GC-MS in the exponential and stationary phases. A total number of 61 VOCs was found in , among which alkanes, alcohols, ketones, and furanic compounds were the most abundant. Some compounds previously reported for (γ-dodecalactone, 3-methylbutan-1-ol, and hexan-1-ol) were also found. Through our experiments, using univariate and multivariate unsupervised (Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA)) and supervised (Projection to Latent Structures Discriminant Analysis (PLS-DA)) statistical techniques, we have proven that each tested growth medium stimulates the production of a different volatiles profile in . Carbon dioxide, hexan-1-ol, pentyl acetate, isomer5 of methyldecane, dimethyl sulphide, undec-5-ene, isomer2 of methylundecane, isomer1 of methyldecane, and 2-methyltetrahydrofuran were established as differentiating compounds among treatments by all the techniques. The significance of our findings deserves future research to investigate if certain volatile profiles could be related to the beneficial or pathogenic role of this yeast.

Keyword: microbiome

Impact of dietary dairy polar lipids on lipid metabolism of mice fed a high-fat diet.

The effect of milk polar lipids on lipid metabolism of liver, adipose tissue, and brain and on composition of intestinal was investigated. C57BL/6J mice were fed a high-fat diet (HFD) for 5 weeks, followed by 5 weeks with HFD without (control) or supplemented with total polar lipids (TPL), phospholipids (PL), or sphingolipids (SPL). Animals fed SPL showed a tendency for lower triglyceride synthesis (P = 0.058) in the liver, but not in adipose tissue. PL and TPL reduced de novo hepatic fatty biosynthesis. The ratio of palmitoleic to in the liver was lower for animals fed SPL or TPL compared to control. There was little effect of the supplementation on the cecal composition. In the brain, DHA (C22:6) content correlated negatively with tetracosanoic (C24:0) after TPL supplementation (-0.71, P = 0.02) but not in control (0.26, P = 0.44). Arachidonic (C20:4) was negatively correlated with C24:0 in both groups (TPL, -0.77, P = 0.008; control, -0.81, P = 0.003).

Keyword: microbiome

Brain and liver fatty composition changes upon consumption of Lactobacillus rhamnosus LA68.

Recent reports suggest that the metabolic activity of the enteric may influence the fatty composition of the host tissue. There are many studies dealing with the influence of lactobacilli on various pathological conditions, and some of the effects are strain-specific. This study was designed to test the effects of a particular Lactobacillus strain, Lactobacillus rhamnosus LA68 on fatty composition of the liver and the brain of C57BL/6 mice in the absence of an underlying pathological condition. Female mice were supplemented with live L. rhamnosus LA68 bacteria for the duration of 1 month. Serum biochemistry was analyzed and liver and brain fatty composition was assessed by gas-liquid chromatography. Significant changes in liver and brain fatty composition were detected. In the liver tissue we detected an increase in palmitoleic (p\u2009=\u20090.038), while in the brain compartment we found an increase in (p\u2009=\u20090.042), stearic (p\u2009=\u20090.017), arachidonic (p\u2009=\u20090.009) and docosahexaenoic (p\u2009=\u20090.004) for control versus experimental group. These results show discrete changes caused by LA68 strain consumption. Even short duration of administration of LA68 influences the fatty composition of the host which adds to the existing knowledge about Lactobacillus host interaction, and adds to the growing knowledge of metabolic intervention possibilities.

Keyword: microbiome

Palm Oil and Beta-palmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition.

Palm oil (PO) is used in infant formulas in order to achieve (PA) levels similar to those in human milk. PA in PO is esterified predominantly at the SN-1,3 position of triacylglycerol (TAG), and infant formulas are now available in which a greater proportion of PA is in the SN-2 position (typical configuration in human milk). As there are some concerns about the use of PO, we aimed to review literature on health effects of PO and SN-2-palmitate in infant formulas.PubMed and Cochrane Database of Systematic Reviews were systematically searched for relevant studies on possible beneficial effects or harms of either PO or SN-2-palmitate in infant formula on various health outcomes.We identified 12 relevant studies using PO and 21 studies using SN-2-palmitate. Published studies have variable methodology, subject characteristics, and some are underpowered for the key outcomes. PO is associated with harder stools and SN-2-palmitate use may lead to softer stool consistency. Bone effects seem to be short-lasting. For some outcomes (infant colic, faecal , lipid metabolism), the number of studies is very limited and summary evidence inconclusive. Growth of infants is not influenced. There are no studies published on the effect on markers of later diseases.There is insufficient evidence to suggest that PO should be avoided as a source of fat in infant formulas for health reasons. Inclusion of high SN-2-palmitate fat blend in infant formulas may have short-term effects on stool consistency but cannot be considered essential.

Keyword: microbiome

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: microbiome

Effect of pasture versus indoor feeding systems on quality characteristics, nutritional composition, and sensory and volatile properties of full-fat Cheddar cheese.

The purpose of this study was to investigate the effects of pasture-based versus indoor total mixed ration (TMR) feeding systems on the chemical composition, quality characteristics, and sensory properties of full-fat Cheddar cheeses. Fifty-four multiparous and primiparous Friesian cows were divided into 3 groups (n = 18) for an entire lactation. Group 1 was housed indoors and fed a TMR diet of grass silage, maize silage, and concentrates; group 2 was maintained outdoors on perennial ryegrass only pasture (GRS); and group 3 was maintained outdoors on perennial ryegrass/white clover pasture (CLV). Full-fat Cheddar cheeses were manufactured in triplicate at pilot scale from each feeding system in September 2015 and were examined over a 270-d ripening period at 8°C. Pasture-derived feeding systems were shown to produce Cheddar cheeses yellower in color than that of TMR, which was positively correlated with increased cheese β-carotene content. Feeding system had a significant effect on the fatty composition of the cheeses. The nutritional composition of Cheddar cheese was improved through pasture-based feeding systems, with significantly lower thrombogenicity index scores and a greater than 2-fold increase in the concentration of vaccenic and the bioactive conjugated linoleic C18:2 cis-9,trans-11, whereas TMR-derived cheeses had significantly higher content. Fatty profiling of cheeses coupled with multivariate analysis showed clear separation of Cheddar cheeses derived from pasture-based diets (GRS or CLV) from that of a TMR system. Such alterations in the fatty profile resulted in pasture-derived cheeses having reduced hardness scores at room temperature. Feeding system and ripening time had a significant effect on the volatile profile of the Cheddar cheeses. Pasture-derived Cheddar cheeses had significantly higher concentrations of the hydrocarbon toluene, whereas TMR-derived cheese had significantly higher concentration of 2,3-butanediol. Ripening period resulted in significant alterations to cheese volatile profiles, with increases in -, alcohol-, aldehyde-, ester-, and terpene-based volatile compounds. This study has demonstrated the benefits of pasture-derived feeding systems for production of Cheddar cheeses with enhanced nutritional and rheological quality compared with a TMR feeding system.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: microbiome

Antioxidant and antidiabetic activity of blackberry after gastrointestinal digestion and human gut fermentation.

Blackberry fruit contains high levels of polyphenols particularly anthocyanins which contribute to its biological activities. Bioavailability of polyphenols especially anthocyanins is generally low, it has been proposed that metabolites from polyphenol biotransformation under colonic fermentation are components that exert health benefits. In this study, blackberry was subjected to simulated gastrointestinal digestion and gut fermentation at different time intervals (0-48\u202fh) to study the changes in bioactive components, its antioxidant and antidiabetic activities. Phenolic compounds, during digestion and fermentation were also analysed. Gut metabolites of blackberry significantly increased the glucose consumption and glycogen content in HepG2 cells. Furthermore, gut metabolites ameliorated high glucose plus -induced ROS overproduction, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. The mechanism of antidiabetic activity of blackberry was via its potent antioxidant activity. Therefore, our results suggest that blackberry could be recommended as a functional food due to potential antioxidant and antidiabetic activity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiome

The Hepatotoxicity of in Zebrafish Involves the Intestinal .

(PA) is the main saturated fatty naturally occurring in animal fats and vegetable oils. In recent decades, palm oil, an alternative lipid source containing high amounts of PA, has been widely used to replace fish oil in aquafeed.We investigated the hepatotoxicity of PA in zebrafish and the underlying mechanism.One-month-old zebrafish fed a high-fat diet (HFD) containing 16% soybean oil and 3 PA-incorporated HFDs [4%, 8%, and 12% PA (12PA)] for 2 wk (experiment 1) and 4 wk (experiment 2) were used to evaluate PA-induced liver damage and endoplasmic reticulum (ER) stress. Germ-free (GF) zebrafish fed low-fat, high-fat, or 12PA diets for 5 d were used to study the direct effects of PA on liver damage (experiment 3). GF zebrafish colonized with HFD or 12PA for 48 h were used to elucidate the indirect effects of PA-altered on liver damage (experiment 4). Last, GF zebrafish colonized with HFD or 12PA were used to evaluate the effects of different microbiotas on PA absorption (experiment 5).In experiment 1, the proportion of PA in the liver linearly increased as its percentage in dietary lipid increased (r2\xa0=\xa00.83, P\xa0<\xa00.05). In experiment 2, the expression of glucose-regulated protein 78 (Grp78) and C/EBP-homologous protein (Chop) was higher in the 12PA group than in the HFD group (2.2- and 2.7-fold, respectively; P\xa0<\xa00.05). The activity of caspase-12 was increased by 61.1% in the 12PA group compared with the HFD group (P\xa0<\xa00.05). In experiment 3, caspase-12 activity was higher in the 12PA group than in the HFD group (P\xa0<\xa00.05). In experiment 4, GF zebrafish colonized with PA-altered had higher caspase-12 activity (P\xa0<\xa00.05) than those colonized by HFD . In experiment 5, PA-altered promoted PA absorption (P\xa0<\xa00.05) and aggravated ER stress and liver damage in the context of high-PA feeding.The PA-altered indirectly induced ER stress and liver damage in zebrafish. Moreover, the PA promoted the absorption of PA, leading to enhanced PA overflow into the liver and aggravated hepatotoxicity of PA in zebrafish.

Keyword: microbiome

Bioremediation and lipid synthesis through mixotrophic algal consortia in municipal wastewater.

Algae grown in outdoor reactors (volume: 10 L and depth: 20 cm) were fed directly with filtered and sterilised municipal wastewater. The nutrient removal efficiencies were 86%, 90%, 89%, 70% and 76% for TOC, TN, NH4-N, TP and OP, respectively, and lipid content varied from 18% to 28.5% of dry algal biomass. Biomass productivity of ∼122 mg/l/d (surface productivity 24.4 g/m(2)/d) and lipid productivity of ∼32 mg/l/d were recorded. Gas chromatography and mass spectrometry (GC-MS) analyses of the fatty methyl esters (FAME) showed a higher content of desirable fatty acids (bearing biofuel properties) with major contributions from saturates such as [C16:0; ∼40%] and stearic [C18:0; ∼34%], followed by unsaturates such as oleic [C18:1(9); ∼10%] and linoleic [C18:2(9,12); ∼5%]. The decomposition of algal biomass and reactor residues with an exothermic heat content of 123.4 J/g provides the scope for further energy derivation.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: microbiome

Bacterial and Metabolic Character of Traditional Sour Cream and Butter in Buryatia, Russia.

Traditional sour cream and butter are widely popular fermented dairy products in Russia for their flavor and nutrition, and contain rich microbial biodiversity, particularly in terms of lactic bacteria (LAB). However, few studies have described the microbial communities and metabolic character of traditional sour cream and butter. The objective of this study was to determine the bacterial and metabolic character of eight samples collected from herdsmen in Buryatia, Russia. Using single-molecule real-time (SMRT) sequencing techniques, we identified a total of 294 species and/or subspecies in 169 bacterial genera, belonging to 14 phyla. The dominant phylum was Firmicutes (81.47%) and the dominant genus was (59.28%). There were differences between the bacterial compositions of the sour cream and butter samples. The relative abundances of , , and were significantly higher in sour cream than in butter, and the abundance of was significantly lower in sour cream than in butter. Using a pure culture method, 48 strains were isolated and identified to represent seven genera and 15 species and/or subspecies. Among these isolates, subsp. (22.50%) was the dominant LAB species. Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry at elevated energy was used in combination with statistical methods to detect metabolite differences between traditional sour cream and butter. A total of 27,822 metabolites were detected in all samples, and Lys-Lys, isohexanal, , Leu-Val, and 2\'-deoxycytidine were the most dominant metabolites found in all samples. In addition, 27 significantly different metabolites were detected between the sour cream and butter samples, including short peptides, organic acids, and amino acids. Based on correlation analyses between the most prevalent bacterial species and the main metabolites in sour cream, we conclude that there may be a connection between the dominant LAB species and these metabolites. This study combined omics techniques to analyze the bacterial diversity and metabolic character of traditional sour cream and butter, and we hope that our findings will enrich species resource libraries and provide valuable resources for further research on dairy product flavor.

Keyword: microbiome

Biomass and community structure of the abyssal determined from the ester-linked phospholipids recovered from Venezuela Basin and Puerto Rico Trench sediments.

Extractible phospholipid fatty acids of abyssal sediment cores from three stations in the Venezuela Basin, transects between them, and a station in the Puerto Rico Trench were analyzed to determine microbial biomass and community composition. Results were compared to abyssal sediments from an area of high-energy boundary currents in the North Atlantic, and estuarine sediments from Apalachee Bay, Florida. Venezuela Basin and Puerto Rico Trench sediments were characterize by low microbial biomass, measured as phospholipid . Venezuela Basin sediments of three different sedimentary regimes showed a remarkably similar microbial community structure, as characterized by fatty profiles. Prokaryotic organisms dominated the microbial community, and fatty acids believed to be signatures of anaerobic organisms were present in greater proportions in Venezuela Basin and Puerto Rico Trench sediments than in either the North Atlantic abyssal sediments or shallow-water estuarine sediments.

Keyword: microbiome

Utility of high performance liquid chromatography/electrospray/mass spectrometry of polar lipids in specifically Per-13C labeled Gram-negative bacteria DA001 as a tracer for acceleration of bioremediation in the subsurface.

Specific fatty acids from phosphatidylglycerol (PG) and phosphatidylethanolamine (PE) recovered from a per 13C-labeled bacteria can be detected in environmental samples and used as measures of bacterial transport in the subsurface. Detection of (16:0) and oleic (18:1) at m/z 271 (255+16) and 299 (281+18) as negative ions in PG and PE separated by high performance liquid chromatography (HPLC) and detected after up-front collisionally induced dissociation (CID) utilizing electrospray (ES) mass spectrometry (MS) provided sufficient sensitivity and specificity for detection in the presence of the indigenous . Application of tandem mass spectrometry (MS/MS) in the multiple reaction monitoring (MRM) was use to monitor selected transitions. MRM can increase the sensitivity so that polar lipids recovered from cell densities currently at about 10(4) cells/sample can be detected. This technology provides a non-intrusive mechanism for monitoring the distribution of bacteria added to accelerate in situ bioremediation of subsurface sediments.

Keyword: microbiome

Obesity changes the human gut mycobiome.

The human intestine is home to a diverse range of bacterial and fungal species, forming an ecological community that contributes to normal physiology and disease susceptibility. Here, the fungal (mycobiome) in obese and non-obese subjects was characterized using Internal Transcribed Spacer (ITS)-based sequencing. The results demonstrate that obese patients could be discriminated by their specific fungal composition, which also distinguished metabolically "healthy" from "unhealthy" obesity. Clusters according to genus abundance co-segregated with body fatness, fasting triglycerides and HDL-cholesterol. A preliminary link to metabolites such as hexadecanedioic , caproic and N-acetyl-L-glutamic was also found. Mucor racemosus and M. fuscus were the species more represented in non-obese subjects compared to obese counterparts. Interestingly, the decreased relative abundance of the Mucor genus in obese subjects was reversible upon weight loss. Collectively, these findings suggest that manipulation of gut mycobiome communities might be a novel target in the treatment of obesity.

Keyword: microbiome

Antioxidant potential and phenolic profile of blackberry anthocyanin extract followed by human gut fermentation.

Beneficial properties attributed to the intake of blackberry fruit are associated with the presence of high content of anthocyanins. However, their low absorption and accumulation in the gut have generated the belief that gut metabolites of anthocyanins are probably reason for their protective effects. In this study, blackberry anthocyanins were prepared and subjected to in vitro human gut fermentation at different time intervals (0-48\u202fh) to study their gut metabolites and antioxidant properties. The content of cyanidin-3-O-glucoside was found highest in blackberry and it degraded completely after 6\u202fh fermentation. Gut metabolites of blackberry anthocyanins were found to improve the glucose consumption and glycogen content significantly in HepG2 cells. Furthermore, gut metabolites significantly ameliorated high glucose plus (HG\u202f+\u202fPA)-induced ROS, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. Overall, this study reveals that blackberry anthocyanins subjected to gut fermentation resulted in the formation of active metabolites with potential antioxidant activity against HG\u202f+\u202fPA-induced oxidative stress.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiome

Alterations to the -colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. -colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut , colonic inflammation and cognitive function to elucidate the role of -gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut . DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut , colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: microbiome

Structure-function relationship of the milk fat globule.

The complex biochemical composition and physical structure of the milk fat globule (MFG) are presented as a basis for its paradoxical metabolic fate: MFG is a rapid conveyor of energy through its triacylglycerol (TAG) core but contains some low-digestible bioactive complex lipids and proteins, which influence lipid metabolism and contribute to intestinal and systemic health.MFG structure modulates gastrointestinal lipolysis, postprandial lipemia and even the postprandial fate of ingested fatty acids. Proof-of-concept of the nutritional programming induced by early consumption of an emulsion biomimetic of MFG compared with a typical infant formula was published in an animal model (mice). The metabolic response to a high-fat diet during adulthood was improved following neonatal exposure to the biomimetic emulsion.MFG TAG are tailored with a unique regiodistribution delivering in priority short to medium-chain fatty acids in gastric phase, an important amount of quickly metabolizable oleic and protecting in sn-2 position. MFG digestion may not only trigger rapid TAG and chylomicron plasma peaks with fast clearance but also the luminal release of nonhydrolysable bioactive compounds (glycosylated compounds and sphingomyelin), which contribute to intestinal and systemic health by shaping the and modulating the immune system. These bioactive compounds form self-assembled structures, protect specific micronutrients and lower cholesterol absorption. The health benefits of MFG consumption or of some of its fractions (MFGM) under specific structures are steadily being demonstrated with still much unsolved questions especially for populations with high nutritional needs (e.g. elderly, infants).

Keyword: microbiome

Exploring the interactions between serum free fatty acids and fecal in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut is involved in the host energy metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal seems to exist. Our aim was to explore the interaction among serum FFA levels, gut , diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiome

Augmented hepatic Toll-like receptors by fatty acids trigger the pro-inflammatory state of non-alcoholic fatty liver disease in mice.

There is considerable evidence that intestinal are involved in the development of metabolic syndromes and, consequently, with the development of non-alcoholic fatty liver disease (NAFLD). Toll-like receptors (TLRs) are essential for the recognition of . However, the induction mechanism of TLR signals through the gut-liver axis for triggering the development of non-alcoholic steatohepatitis (NASH) or NAFLD remains unclear. In this study, we investigated the role of (PA) in triggering the development of a pro-inflammatory state of NAFLD.Non-alcoholic fatty liver disease was induced in mice fed a high fat diet (HFD). The mice were killed and the expression of TLRs, tumor necrosis factor (TNF), interleukin (IL)-1β, and phospho-interleukin-1 receptor-associated kinase 1 in the liver and small intestine were assessed. In addition, primary hepatocytes and Kupffer cells were treated with PA, and the direct effects of PA on TLRs induction by these cells were evaluated.The expression of inflammatory cytokines such as TNF, IL-1β, and TLR-2, -4, -5, and -9 was increased in the liver, but decreased in the small intestine of HFD-fed mice in vivo. In addition, the expression of TLRs in primary hepatocytes and Kupffer cells was increased by treatment with PA.In the development of the pro-inflammatory state of NAFLD, PA triggers the expression of TLRs, which contribute to the induction of inflammatory cytokines through TLR signals by intestinal .© 2013 The Japan Society of Hepatology.

Keyword: microbiome

Gut and Metabolome Response of Seed Oil on Metabolism Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.

Keyword: microbiome

Effect of high β-palmitate content in infant formula on the intestinal of term infants.

(PA) constitutes 17% to 25% of the human milk fatty acids, and ~70% is esterified in the sn-2 position of triglycerides (β-palmitate). In the sn-2 position, PA is not hydrolyzed and thus is efficiently absorbed. The PA in palm oils, commonly used in infant formulas, is esterified in the sn-1 and sn-3 positions. In these positions, PA is hydrolyzed and forms poorly absorbed calcium complexes. The present study assessed whether high β-palmitate in infant formulas affects the intestinal flora.Thirty-six term infants were enrolled: 14 breast-fed (BF group) and 22 formula-fed infants who were randomly assigned to receive formula containing high β-palmitate (HBP group, n=14), or low β-palmitate (LBP group, n=8), where 44% and 14% of the PA was β-palmitate, respectively. The total amount of PA in the formulas was 19% and 22% in the LBP and HBP groups, respectively. Neither formula contained pre- or probiotics. Stool samples were collected at enrollment and at 6 weeks for the quantification of bacteria.At 6 weeks, the HBP and BF groups had higher Lactobacillus and bifidobacteria counts than the LBP group (P<0.01). The Lactobacillus counts at 6 weeks were not significantly different between the HBP and BF groups. Lactobacillus counts were 1.2×10¹⁰, 1.2×10¹¹, and 5.6×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively. Bifidobacteria counts were 5.1×10⁹, 1.2×10¹¹, and 3.9×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively.HBP formula beneficially affected infant gut by increasing the Lactobacillus and bifidobacteria counts in fecal stools.ClinicalTrials.gov .

Keyword: microbiome

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines metabolism, and gut microbiome-derived metabolism. Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested fat decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: microbiome

[The oleic triglycerides of palm oil and triglycerides of creamy fat. The reaction of palmitoylation, potassium and magnesium palmitate, absorption of fatty acids by enterocytes and of large intestine].

The decreasing of content of animal, milk fat (butter) by means of its substitution with vegetable, oleic, palmy oil in food of adults optimal by its quantity is physically chemically and biologically substantiated. In oleic palmy oil higher content of oleic mono unsaturated fatty and oleic triglycerides than in creamy fat is established. The biologic availability of unsaturated in the form of free fatty is decreased at its absorption by enterocytes of small intestines is detected. There are no transforms of mono unsaturated acids in palmy oil in contrast with hydrogenated margarines. In palmy, oleic oil there is not enough of short-chained fatty acids (C4-C6) and it has no taste quality and it has low level of unsaturated fatty acids and factually it is lacking of ω-6 polyunsaturated fatty acids. However, it is compensated in case of availability offish and sea products in food. If adults, especially older ones, will refuse to consume creamy fat and decrease intake of products with high content of unsaturated fatty and triglycerides (beef, sour cream, fatty cheeses) it\'ll positively impact their health. The refusal from these products is a real step in prevention of metabolic pandemic (atherosclerosis and atheromatosis, metabolic syndrome, resistance to insulin, obesity). There are still large number of people who at optimal amount of food retain in vivo increased amount of exogenous, endogenously synthesized from glucose unsaturated fatty in the form of unesterified fatty acids (syndrome of resistance to insulin) and increased content of triglycerides.

Keyword: microbiome

Rhizosphere effect of Scirpus triqueter on soil microbial structure during phytoremediation of diesel-contaminated wetland.

Though phytoremediation has been widely used to restore various contaminated sites, it is still unclear how soil microbial communities respond microecologically to plants and pollutants during the process. In this paper, batch microcosms imitating in situ phytoremediation of petroleum-contaminated wetland by Scirpus triqueter were set up to monitor the influence of plant rhizosphere effect on soil microbes. , one of the main root exudates of S. triqueter, was added to strengthen rhizosphere effect. Abundances of certain microbial subgroups were quantified by phospholipid fatty profiles. Results showed that diesel removal extents were significantly higher in the rhizosphere (57.6 +/-4.2-65.5 +/- 6.9%) than those in bulk soil (27.8 +/-6.5-36.3 +/- 3.2%). In addition, abundances of saturated, monounsaturated, and polyunsaturated fatty acids were significantly higher (P < 0.05) in planted soil than those in the bulk soil. When it was less than 15,000 mg diesel kg soil-1, increasing diesel concentration led to higher abundances of fungi, Gram-positive and Gram-negative bacteria. The addition of amplified the rhizosphere effect on soil microbial populations and diesel removal. Principal component analysis revealed that plant rhizosphere effect was the dominant factor affecting microbial structure. These results provided new insights into plant-microbe-pollutant coactions responsible for diesel degradation, and they were valuable to facilitate phytoremediation of diesel contamination in wetland habitats.

Keyword: microbiome

Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut , and hepatic metabolism.

Keyword: microbiome

Regulation of -GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of -GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: microbiome

Effect of silicate grain shape, structure, and location on the biomass and community structure of colonizing marine .

colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable . With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined.

Keyword: microbiome

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows.

Increasing the dietary fat content of ruminant diets decreases methane (CH) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h)] consisting of 12.1% , 4.2% stearic , 31.1% oleic , and 52.7% linoleic . Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH production and DMI were lower, whereas daily CH yield (CH/DMI) was greater in LIPO than CON cows, although CH yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: microbiome

Tuning culturing conditions towards the production of neutral lipids from lubricant-based wastewater in open mixed bacterial communities.

Production of bacterial lipid-based biofuels using inexpensive substrates, as wastes, is an emerging approach. In this work, a selective process using carbon feast-famine cycles was applied to obtain an indigenous microbial community of hydrocarbon-degrading and lipid-accumulating bacteria, using a real lubricant-based wastewater as carbon source. In the conditions applied, the enriched bacterial community, dominated by members of the genus Rhodococcus, Pseudomonas and Acinetobacter, was able to degrade almost all hydrocarbons present in the wastewater within 24\u202fh\' incubation and to accumulate, although in low levels, triacylglycerol (TAG) (<5% of cell dry weight (CDW)) and polyhydroxyalkanoates (PHA) (3.8%\u202f±\u202f1.1% of the CDW) as well as an unknown lipid (29%\u202f±\u202f6% of CDW), presumably a wax ester-like compound. The influence of culture conditions, namely carbon and nitrogen concentrations (and C/N ratio) and cultivation time, on the amount and profile of produced storage compounds was further assessed using a statistical approach based on a central composite circumscribed design and surface response methodology. The regression analysis of the experimental design revealed that only nitrogen concentration and C/N ratio are significant for neutral lipid biosynthesis (p\u202f<\u202f0.05). Maximum neutral lipid content, i.e. 33% (CDW basis), was achieved for the lowest carbon and nitrogen concentrations evaluated (10\u202fg COD L and 0.02\u202fg\u202fN L). PHA accounted for less than 5% of CDW. In these conditions, neutral lipid content was mainly composed by TAG, about 70% (w/w). TAG precursors, namely monoacylglycerols (MAG), diacylglycerols (DAG) and fatty acids (FA), accounted for 22% of total neutral lipids and WE for about 7%. Nevertheless, according to the applied response surface model, further improvement of neutral lipids content is still possible if even lower nitrogen concentrations are used. The fatty acids detected in TAG extracts ranged from myristic (C14:0) to linoleic (C18:2), being the most abundant (C16:0), stearic (C18:0) and oleic (C18:1). This study shows the feasibility of combining treatment of hydrocarbon contaminated wastewater, herein demonstrated for lubricant-based wastewater, with the production of bacterial neutral lipids using open mixed bacterial communities. This approach can decrease the costs associated to both processes and contribute to a more sustainable waste management and production of lipid-based biofuels.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiome

Association between sn-2 fatty profiles of breast milk and development of the infant intestinal microbiome.

Increasing evidence shows that host diet and gut microbes are related. Previous studies have shown the effects of specific dietary fatty acids (FAs) on intestinal , but little is known about the effect of the stereospecifically numbered sn-2 position in triglycerides (TG) of human milk on the gut microbiome of infants. This study aimed at examining possible effects of sn-2 FAs of human milk on the gut microbial development of breastfeeding babies. Sn-2 FAs and intestinal were assessed by GC-MS and high-throughput 16S rRNA sequencing, respectively. The results showed that breast milk from mothers in China contained ten major sn-2 FAs dominated by (C, 54.42%), oleic (C n-9, 14.95%), linoleic (LA, C n-6, 12.81%), myristic (C, 4.50%) and C (3.17%). Total long chain unsaturated fatty acids (LCUFA) decreased from colostrum to mature milk, while total saturated fatty acids (SFA) showed no significant difference during lactation. A significant association between sn-2 FAs in milk and infant gut was found between decanoic (C), myristic (C), stearic (C), C, arachidonic (AA, C n-6), docosahexaenoic (DHA, C n-3) with Bacteroides, Enterobacteriaceae, Veillonella, Streptococcus, and Clostridium. These microbes were involved in short-chain fatty (SCFA) production and other functions, and significantly increased at 13-15 d after breastfeeding was initiated. C and DHA were relevant to most of the microbes. This study demonstrated the relatively steady profiles of sn-2 FAs in breast milk and gut of infants, together with their correlation during the breastfeeding period. The above results provided important information for designing the configuration of FAs in next-generation formulas for Chinese infants.

Keyword: microbiome

Metabolic pathway and role of individual species in the bacterial consortium for biodegradation of azo dye: A biocalorimetric investigation.

In this study, an attempt was made to investigate the functional role and metabolic behaviour of the monoculture (Staphylococcus lentus (SL), Bacillus flexus (BF) and Pseudomonas aeruginosa (PA)) in the bacterial biocenosis for biotransformation of an azo dye. The power-time profile obtained from consortia depicted three distinct peaks, which correlated well with the individual bacterial growth (PA\xa0>\xa0SL\xa0>\xa0BF), indicating the synergistic relation and division of labour in the biocenosis. The heat release pattern was used to identify the sequential behaviour of microbial consortia in real time. Yield calculation based on total heat liberated to the complete substrate utilization Y for PA, SL, and BF were 15.99, 16.68, 7.32\xa0kJ/L respectively. Similarly, the oxy calorific values Y for the above species are respectively 386, 375, 440\xa0kJ/mol and indicates the aerobic nature of microorganism employed. Further, the metabolome produced during the biotransformation were identified using Gas Chromatography-Mass Spectrometry (GC-MS), based on which a plausible pathway was predicted. The abundant metabolites were (m/z\xa0=\xa0256) and diethyl phthalate (m/z\xa0=\xa0222.2). The abundance of diethyl phthalate was much lesser in the consortia compared to the monoculture. Thus, the biocalorimetric heat yield calculation along with the stoichiometry and plausible pathway based biochemical elucidation provides a mechanistic basis for understanding the azo-dye biotransformation by the monocultures in consortia.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: microbiome

Mode of action of a fatty -based natural product to control Botrytis cinerea in grapes.

To investigate the efficacy and mode of action of the fatty -based product Foodcoat(®) (FC) against Botrytis cinerea.In vitro, in vivo and field experiments were carried out to investigate the effect of different concentrations of FC on B. cinerea germination and infection of grape leaves and berries, using three selected isolates and comparing results with those achieved by the commercialized product Protector(HML) (PRT). Furthermore, the effect of field applications of FC on the grape berry was investigated. FC reduced B. cinerea germination and grape berry severity by up to 54 and 96%, respectively, compared with the untreated controls.Foodcoat demonstrated efficacy that was equal or greater than the registered product, PRT. A multiple mode of action was hypothesized for FC suppression of B. cinerea, including: inhibition of germination and germ tube alteration, protection of host green tissues and enhancement of the natural yeast populations on the berry surface.The efficacy of both products has been quantified and their modes of action described, suggesting them for field applications against B. cinerea, alone or in combined strategies. This is also the first report of a fatty -based product stimulating natural yeast populations on grape berries.© 2014 The Society for Applied Microbiology.

Keyword: microbiome

Characterization of aerobic oil and grease-degrading bacteria in wastewater.

A bacterial consortium that degrades cooking oil (CO) has been isolated in wastewater (WW) samples, by enrichment in olive CO. This consortium could degrade 90% of CO within 7-9 days (from an initial 1% [w/v]), and it is more active at alkaline conditions. The 16S ribonucleic (RNA) gene analysis showed that it contains five bacterium species: Stenotrophomonas rhizophila, Sphingobacterium sp., Pseudomonas libanensis, Pseudomonas poae and Pseudomonas aeruginosa. This consortium can degrade the free fatty acids (FFA): , stearic, oleic, linoleic and linolenic acids; glycerol, glucose and amylose; and albumin, but could not efficiently degrade carboxymethyl-cellulose. Each strain could also degrade CO and FFAs. The level of bacterial crude-activity of extracellular lipases was found to be between 0.2 and 4U/ml. Using synthetic WW, the consortium could reduce 80% of the chemical oxygen demand [from 10550\u2009±\u20092828\u2005mg/l], 80% of nitrogen (from 410\u2009±\u200978\u2009mgl/l) and 57% of phosphorus (from 93\u2009±\u200925\u2005mg/l). Thus, this consortium can be utilized in the removal of CO from WW.

Keyword: microbiome

Gut -mediated generation of saturated fatty acids elicits inflammation in the liver in murine high-fat diet-induced steatohepatitis.

The gut plays crucial roles in the development of non-alcoholic steatohepatitis (NASH). However, the precise mechanisms by which alterations of the gut and its metabolism contributing to the pathogenesis of NASH are not yet fully elucidated.Mice were fed with a recently reported new class of high-fat diet (HFD), steatohepatitis-inducing HFD (STHD)-01 for 9\xa0weeks. The composition of the gut was analyzed by T-RFLP. Luminal metabolome was analyzed using capillary electrophoresis and liquid chromatography time-of-flight mass spectrometry (CE- and LC-TOFMS).Mice fed the STHD-01 developed NASH-like pathology within a short period. Treatment with antibiotics prevented the development of NASH by STHD-01. The composition of the gut and its metabolic activities were markedly perturbed in the STHD-01-fed mice, and antibiotic administration normalized these changes. We identified that long-chain saturated fatty and n-6 fatty metabolic pathways were significantly altered by STHD-01. Of note, the changes in gut lipidome caused by STHD-01 were mediated by gut , as the depletion of the gut could reverse the perturbation of these metabolic pathways. A saturated long-chain fatty , , which accumulated in the STHD-01 group, activated liver macrophages and promoted TNF-α expression.Lipid metabolism by the gut , particularly the saturation of fatty acids, affects fat accumulation in the liver and subsequent liver inflammation in NASH.

Keyword: microbiome

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile Metabolism and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut , is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) metabolism. However, whether TMAO aggravates liver steatosis by modulating BA metabolism and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA metabolism. Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: microbiome

Beta-palmitate - a natural component of human milk in supplemental milk formulas.

The composition and function of human milk is unique and gives a basis for the development of modern artificial milk formulas that can provide an appropriate substitute for non-breastfed infants. Although human milk is not fully substitutable, modern milk formulas are attempting to mimic human milk and partially substitute its complex biological positive effects on infants. Besides the immunomodulatory factors from human milk, research has been focused on the composition and structure of human milk fat with a high content of β- (sn-2 , β-palmitate). According to the available studies, increasing the content of β-palmitate added to milk formulas promotes several beneficial physiological functions. β-palmitate positively influences fatty metabolism, increases calcium absorption, improves bone matrix quality and the stool consistency, and has a positive effect on the development of the intestinal microbiome.

Keyword: microbiome

Characterisation of Fecal Soap Fatty Acids, Calcium Contents, Bacterial Community and Short-Chain Fatty Acids in Sprague Dawley Rats Fed with Different sn-2 Triacylglycerols Diets.

The structure of dietary triacylglycerols is thought to influence fatty and calcium absorption, as well as intestinal population of the host. In the present study, we investigated the impact of (PA) esterified at the sn-2 position on absorption of fatty and calcium and composition of intestinal microorganisms in rats fed high-fat diets containing either low sn-2 PA (12.1%), medium sn-2 PA (40.4%) or high sn-2 PA (56.3%), respectively. Fecal fatty profiles in the soaps were measured by gas chromatography (GC), while fecal calcium concentration was detected by ICP-MS. The fecal microbial composition was assessed using a 16S rRNA high-throughput sequencing technology and fecal short-chain fatty acids were detected by ion chromatograph. Dietary supplementation with a high sn-2 PA fat significantly reduced total fecal contents of fatty acids soap and calcium compared with the medium or low sn-2 PA fat groups. Diet supplementation with sn-2 PA fat did not change the entire profile of the gut community at phylum level and the difference at genera level also were minimal in the three treatment groups. However, high sn-2 PA fat diet could potentially improve total short-chain fatty acids content in the feces, suggesting that high dietary sn-2 PA fat might have a beneficial effect on host intestinal health.

Keyword: microbiome

Lipid Metabolic Versatility in spp. Yeasts Studied through Metabolic Modeling.

species are lipophilic and lipid-dependent yeasts belonging to the human and animal . Typically, they are isolated from regions rich in sebaceous glands. They have been associated with dermatological diseases such as seborrheic dermatitis, pityriasis versicolor, atopic dermatitis, and folliculitis. The genomes of , , and lack the genes related to fatty synthesis. Here, the lipid-synthesis pathways of these species, as well as of , and of an atypical variant were reconstructed using genome data and Constraints Based Reconstruction and Analysis. To this end, the genomes of CBS 1878 and the atypical 4DS were sequenced and annotated. The resulting Enzyme Commission numbers and predicted reactions were similar to the other strains despite the differences in their genome size. Proteomic profiling was utilized to validate flux distributions. Flux differences were observed in the production of steroids in and in the metabolism of butanoate in . The predictions obtained via these metabolic reconstructions also suggested defects in the assimilation of in , , , and the atypical variant of , but not in These predictions were validated via physiological characterization, showing the predictive power of metabolic network reconstructions to provide new clues about the metabolic versatility of .

Keyword: microbiome

Metabolomics reveals potential biomarkers in the rumen fluid response to different milk production of dairy cows.

In the present study, an LC/MS metabolomics approach was performed to investigate potential biomarkers of milk production in high- and low-milk-yield dairy cows and to establish correlations among rumen fluid metabolites; the results of this study provide insights into the mechanisms underlying the milk production-related characteristics of rumen fluid in dairy cows. Sixteen lactating dairy cows with similar parity and days in milk were divided into high-yield (HY) and low-yield (LY) groups based on milk yield. On day 21, rumen fluid metabolites were quantified applying LC/MS. The principal component analysis (PCA) and orthogonal partial least-squares (OPLS-DA) showed significantly separated clusters of the ruminal metabolite profiles of HY and LY groups. Compared with HY group, a total of 24 ruminal metabolites were significantly greater in LY group, such as 3-hydroxyanthranilic , carboxylic acids, carboxylic derivatives (L-isoleucine, L-valine, L-tyrosine, etc.), diazines (uracil, thymine, cytosine), and , while the concentrations of 30 metabolites were dramatically decreased in LY group compared to HY group, included gentisic , caprylic , and myristic . The metabolite enrichment analysis indicated that protein digestion and absorption, ABC transporters and unsaturated fatty biosynthesis were significantly different between the two groups. Correlation analysis between the ruminal microbiome and metabolites revealed that certain typical metabolites were exceedingly associated with definite ruminal bacteria; Firmicutes, Actinobacteria and Synergistetes phyla were highly correlated with most metabolites. These findings revealed that the ruminal metabolite profiles were significantly different between HY and LY groups, and these results may provide novel insights to evaluate biomarkers for a better feed digestion and may reveal the potential mechanism underlying the difference in milk yield in dairy cows.

Keyword: microbiome

Dynamic alterations in the gut and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut and metabolome.

Keyword: microbiome

Lipotoxicity and the gut-liver axis in NASH pathogenesis.

The pathogenesis of non-alcoholic fatty liver disease, particularly the mechanisms whereby a minority of patients develop a more severe phenotype characterised by hepatocellular damage, inflammation, and fibrosis is still incompletely understood. Herein, we discuss two pivotal aspects of the pathogenesis of NASH. We first analyse the initial mechanisms responsible for hepatocellular damage and inflammation, which derive from the toxic effects of excess lipids. Accumulating data indicate that the total amount of triglycerides stored in hepatocytes is not the major determinant of lipotoxicity, and that specific lipid classes act as damaging agents on liver cells. In particular, the role of free fatty acids such as , cholesterol, lysophosphatidylcholine and ceramides has recently emerged. These lipotoxic agents affect the cell behaviour via multiple mechanisms, including activation of signalling cascades and death receptors, endoplasmic reticulum stress, modification of mitochondrial function, and oxidative stress. In the second part of this review, the cellular and molecular players involved in the cross-talk between the gut and the liver are considered. These include modifications to the , which provide signals through the intestine and bacterial products, as well as hormones produced in the bowel that affect metabolism at different levels including the liver. Finally, the activation of nuclear receptors by bile acids is analysed.Copyright © 2017. Published by Elsevier B.V.

Keyword: microbiome

The influence of the position of palmitate in infant formula triacylglycerols on health outcomes.

The purpose of this review is to discuss recent studies reporting on the influence of the position of in triacylglycerols in infant formula and relevant animal studies. Earlier experiments in rodents show that a diet with a higher proportion of palmitate at the sn-2 position of triacylglycerols improves dietary fat and calcium absorption compared with a diet with a lower sn-2 palmitate content. A high-sn-2 palmitate diet increased fecal short-chain fatty acids, reduced gut inflammation in a colitis model, and altered tissue endocannabinoid concentrations in laboratory rodents. Recent studies in infants confirm that formula with a high sn-2 palmitate content reduces stool fat, , fat soaps, palmitate soaps, and calcium compared with formula with a low sn-2 palmitate content. These effects have been associated with improved bone strength, increased fecal bifidobacteria, and reduced crying in infants. In some studies, findings with formula high in sn-2 palmitate match those seen in breast-fed infants. However, in many studies, high sn-2 palmitate formula remains inferior to breast-feeding. It is concluded that infant formula high in sn-2 palmitate is superior to formula with low sn-2 palmitate but does not fully match human breast milk. Recent studies showing altered gut (human infants) and tissue endocannabinoids (rodent model) suggest the potential for marked physiological impact of high sn-2 palmitate that needs to be explored further in human trials.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: microbiome

Time-Dependent alteration to the tight junction structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal composition, increased intestinal permeability, and onset of type 2 diabetes mellitus (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the tight junction (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the , after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: microbiome

Anaerococcus rubiinfantis sp. nov., isolated from the gut of a Senegalese infant with severe acute malnutrition.

Anaerococcus rubiinfantis sp. nov. strain mt16(T) is a new species within the genus Anaerococcus, which was isolated by the culturomics approach from the gut of an infant suffering from kwashiorkor. A phenotypic, biochemical and proteomic description of this strain is hereby presented alongside a complete annotation of its genome. This strictly anaerobic species forms Gram-positive non-sporeforming cocci. The major fatty was hexadecanoic . The phylogenetic analysis of strain mt16(T) showed a 97.9% similarity level with Anaerococcus vaginalis, the closest validly published species. Its genome is 1,929,161\xa0bp long with 29.5% G\xa0+\xa0C content and contains 1808 protein-coding genes and 56 RNA genes, among which are six rRNA genes. Genomic analysis identified 41/1864 coding genes as ORFans (2.2%) and at least 620/1808 (34.9%) orthologous proteins which are not shared with the closest phylogenetic species. We believe that the extension of the human anaerobic gut compendium by culturomics is one of the first steps that will improve the understanding of the links between the microbiome and health or disease.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: microbiome

The effect of saturated and unsaturated fatty acids on the production of outer membrane vesicles from and .

The aim of present study is to investigate the effect of fatty acids on the outer membrane vesicles (OMVs) produced by spp. spp. is the important member of Gut that employ OMVs production for interact with host. Besides, dietary fatty acids could influence on determination of gut composition and immune response. In this regard, we evaluated the effect of fatty acids on the growth and OMVs production of and . and were grown on BHI broth with and without and palmitoleic acids as saturated and unsaturated fatty acids, respectively. OMVs were extracted using multiple centrifugation and tris-ethylene diamine tetra acetic (EDTA)-Sodium deoxy cholate buffers. Physicochemical properties of OMVs were detected by electron microscopy (SEM), Bradford Coomassie brilliant blue assay and SDS-PAGE. Data were analyzed with One-way ANOVA using SPSS.The growths of both were significantly increased by . Nevertheless, palmitoleic had no significant effect on them. significantly decreased and increased the production of OMVs at low and high concentration, respectively. However, the production of OMVs was not significantly affected by . Although palmitoleic had a significant decreasing effect on the production of OMVs, it significantly increased the production of OMVs at low concentration.In conclusion we reported that had a stimulatory effect on the growth of and and had a dose dependent effect on the production of OMVs. Also producing of OMVs was affected by palmitoleic in a dose dependent manner.

Keyword: microbiota

The impact of dietary sn-2 triacylglycerols in combination with docosahexaenoic or arachidonic on lipid metabolism and host faecal composition in Sprague Dawley rats.

Sn-2 triacylglycerols (sn2PA fat) and polyunsaturated fatty acids are thought to influence the metabolic status and intestinal bacterial population of the host. In this study, the impact of sn2PA fat in combination with DHA or ARA in the diet on lipid metabolism in the liver and faecal composition were investigated in rats fed diets containing sn2PA fat, 90% sn2PA fat + 10% DHA oil (wt%), or 90% sn2PA fat + 10% ARA oil (wt%). Tissue fatty composition was measured using gas chromatography (GC), whereas the faecal microbial composition was assessed using 16S rRNA high-throughput sequencing technology. In addition, faecal short-chain fatty acids (SCFA) were analyzed using ion chromatography. The results showed that sn2PA fat in combination with DHA or ARA significantly reduced liver triacylglyceride (TG) content compared with the sn2PA fat only group. Moreover, the supplementation with sn2PA fat in combination with DHA or ARA significantly promoted the growth of Lactobacillus in the feces at the genus level. On the other hand, the growth of the opportunistic pathogen Desulfovibrio was significantly inhibited by sn2PA fat in combination with ARA compared with the sn2PA fat group. In addition, sn2PA fat in combination with DHA or ARA significantly increased total SCFA concentration in the faeces, suggesting a beneficial effect on host intestinal health.

Keyword: microbiota

Yeast Smell Like What They Eat: Analysis of Volatile Organic Compounds of in Growth Media Supplemented with Different Lipids.

is part of the human skin . Its volatile organic compounds (VOCs) possibly contribute to the characteristic odour in humans, as well as to interaction. The aim of this study was to investigate how the lipid composition of the liquid medium influences the production of VOCs. Growth was performed in four media: (1) mDixon, (2) oleic (OA), (3) oleic + (OA+PA), and (4) (PA). The profiles of the VOCs were characterized by HS-SPME/GC-MS in the exponential and stationary phases. A total number of 61 VOCs was found in , among which alkanes, alcohols, ketones, and furanic compounds were the most abundant. Some compounds previously reported for (γ-dodecalactone, 3-methylbutan-1-ol, and hexan-1-ol) were also found. Through our experiments, using univariate and multivariate unsupervised (Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA)) and supervised (Projection to Latent Structures Discriminant Analysis (PLS-DA)) statistical techniques, we have proven that each tested growth medium stimulates the production of a different volatiles profile in . Carbon dioxide, hexan-1-ol, pentyl acetate, isomer5 of methyldecane, dimethyl sulphide, undec-5-ene, isomer2 of methylundecane, isomer1 of methyldecane, and 2-methyltetrahydrofuran were established as differentiating compounds among treatments by all the techniques. The significance of our findings deserves future research to investigate if certain volatile profiles could be related to the beneficial or pathogenic role of this yeast.

Keyword: microbiota

Impact of dietary dairy polar lipids on lipid metabolism of mice fed a high-fat diet.

The effect of milk polar lipids on lipid metabolism of liver, adipose tissue, and brain and on composition of intestinal was investigated. C57BL/6J mice were fed a high-fat diet (HFD) for 5 weeks, followed by 5 weeks with HFD without (control) or supplemented with total polar lipids (TPL), phospholipids (PL), or sphingolipids (SPL). Animals fed SPL showed a tendency for lower triglyceride synthesis (P = 0.058) in the liver, but not in adipose tissue. PL and TPL reduced de novo hepatic fatty biosynthesis. The ratio of palmitoleic to in the liver was lower for animals fed SPL or TPL compared to control. There was little effect of the supplementation on the cecal composition. In the brain, DHA (C22:6) content correlated negatively with tetracosanoic (C24:0) after TPL supplementation (-0.71, P = 0.02) but not in control (0.26, P = 0.44). Arachidonic (C20:4) was negatively correlated with C24:0 in both groups (TPL, -0.77, P = 0.008; control, -0.81, P = 0.003).

Keyword: microbiota

Brain and liver fatty composition changes upon consumption of Lactobacillus rhamnosus LA68.

Recent reports suggest that the metabolic activity of the enteric may influence the fatty composition of the host tissue. There are many studies dealing with the influence of lactobacilli on various pathological conditions, and some of the effects are strain-specific. This study was designed to test the effects of a particular Lactobacillus strain, Lactobacillus rhamnosus LA68 on fatty composition of the liver and the brain of C57BL/6 mice in the absence of an underlying pathological condition. Female mice were supplemented with live L. rhamnosus LA68 bacteria for the duration of 1 month. Serum biochemistry was analyzed and liver and brain fatty composition was assessed by gas-liquid chromatography. Significant changes in liver and brain fatty composition were detected. In the liver tissue we detected an increase in palmitoleic (p\u2009=\u20090.038), while in the brain compartment we found an increase in (p\u2009=\u20090.042), stearic (p\u2009=\u20090.017), arachidonic (p\u2009=\u20090.009) and docosahexaenoic (p\u2009=\u20090.004) for control versus experimental group. These results show discrete changes caused by LA68 strain consumption. Even short duration of administration of LA68 influences the fatty composition of the host which adds to the existing knowledge about Lactobacillus host interaction, and adds to the growing knowledge of metabolic intervention possibilities.

Keyword: microbiota

Palm Oil and Beta-palmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition.

Palm oil (PO) is used in infant formulas in order to achieve (PA) levels similar to those in human milk. PA in PO is esterified predominantly at the SN-1,3 position of triacylglycerol (TAG), and infant formulas are now available in which a greater proportion of PA is in the SN-2 position (typical configuration in human milk). As there are some concerns about the use of PO, we aimed to review literature on health effects of PO and SN-2-palmitate in infant formulas.PubMed and Cochrane Database of Systematic Reviews were systematically searched for relevant studies on possible beneficial effects or harms of either PO or SN-2-palmitate in infant formula on various health outcomes.We identified 12 relevant studies using PO and 21 studies using SN-2-palmitate. Published studies have variable methodology, subject characteristics, and some are underpowered for the key outcomes. PO is associated with harder stools and SN-2-palmitate use may lead to softer stool consistency. Bone effects seem to be short-lasting. For some outcomes (infant colic, faecal , lipid metabolism), the number of studies is very limited and summary evidence inconclusive. Growth of infants is not influenced. There are no studies published on the effect on markers of later diseases.There is insufficient evidence to suggest that PO should be avoided as a source of fat in infant formulas for health reasons. Inclusion of high SN-2-palmitate fat blend in infant formulas may have short-term effects on stool consistency but cannot be considered essential.

Keyword: microbiota

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: microbiota

Antioxidant and antidiabetic activity of blackberry after gastrointestinal digestion and human gut fermentation.

Blackberry fruit contains high levels of polyphenols particularly anthocyanins which contribute to its biological activities. Bioavailability of polyphenols especially anthocyanins is generally low, it has been proposed that metabolites from polyphenol biotransformation under colonic fermentation are components that exert health benefits. In this study, blackberry was subjected to simulated gastrointestinal digestion and gut fermentation at different time intervals (0-48\u202fh) to study the changes in bioactive components, its antioxidant and antidiabetic activities. Phenolic compounds, during digestion and fermentation were also analysed. Gut metabolites of blackberry significantly increased the glucose consumption and glycogen content in HepG2 cells. Furthermore, gut metabolites ameliorated high glucose plus -induced ROS overproduction, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. The mechanism of antidiabetic activity of blackberry was via its potent antioxidant activity. Therefore, our results suggest that blackberry could be recommended as a functional food due to potential antioxidant and antidiabetic activity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiota

The Hepatotoxicity of in Zebrafish Involves the Intestinal .

(PA) is the main saturated fatty naturally occurring in animal fats and vegetable oils. In recent decades, palm oil, an alternative lipid source containing high amounts of PA, has been widely used to replace fish oil in aquafeed.We investigated the hepatotoxicity of PA in zebrafish and the underlying mechanism.One-month-old zebrafish fed a high-fat diet (HFD) containing 16% soybean oil and 3 PA-incorporated HFDs [4%, 8%, and 12% PA (12PA)] for 2 wk (experiment 1) and 4 wk (experiment 2) were used to evaluate PA-induced liver damage and endoplasmic reticulum (ER) stress. Germ-free (GF) zebrafish fed low-fat, high-fat, or 12PA diets for 5 d were used to study the direct effects of PA on liver damage (experiment 3). GF zebrafish colonized with HFD or 12PA for 48 h were used to elucidate the indirect effects of PA-altered on liver damage (experiment 4). Last, GF zebrafish colonized with HFD or 12PA were used to evaluate the effects of different microbiotas on PA absorption (experiment 5).In experiment 1, the proportion of PA in the liver linearly increased as its percentage in dietary lipid increased (r2\xa0=\xa00.83, P\xa0<\xa00.05). In experiment 2, the expression of glucose-regulated protein 78 (Grp78) and C/EBP-homologous protein (Chop) was higher in the 12PA group than in the HFD group (2.2- and 2.7-fold, respectively; P\xa0<\xa00.05). The activity of caspase-12 was increased by 61.1% in the 12PA group compared with the HFD group (P\xa0<\xa00.05). In experiment 3, caspase-12 activity was higher in the 12PA group than in the HFD group (P\xa0<\xa00.05). In experiment 4, GF zebrafish colonized with PA-altered had higher caspase-12 activity (P\xa0<\xa00.05) than those colonized by HFD . In experiment 5, PA-altered promoted PA absorption (P\xa0<\xa00.05) and aggravated ER stress and liver damage in the context of high-PA feeding.The PA-altered indirectly induced ER stress and liver damage in zebrafish. Moreover, the PA promoted the absorption of PA, leading to enhanced PA overflow into the liver and aggravated hepatotoxicity of PA in zebrafish.

Keyword: microbiota

Bioremediation and lipid synthesis through mixotrophic algal consortia in municipal wastewater.

Algae grown in outdoor reactors (volume: 10 L and depth: 20 cm) were fed directly with filtered and sterilised municipal wastewater. The nutrient removal efficiencies were 86%, 90%, 89%, 70% and 76% for TOC, TN, NH4-N, TP and OP, respectively, and lipid content varied from 18% to 28.5% of dry algal biomass. Biomass productivity of ∼122 mg/l/d (surface productivity 24.4 g/m(2)/d) and lipid productivity of ∼32 mg/l/d were recorded. Gas chromatography and mass spectrometry (GC-MS) analyses of the fatty methyl esters (FAME) showed a higher content of desirable fatty acids (bearing biofuel properties) with major contributions from saturates such as [C16:0; ∼40%] and stearic [C18:0; ∼34%], followed by unsaturates such as oleic [C18:1(9); ∼10%] and linoleic [C18:2(9,12); ∼5%]. The decomposition of algal biomass and reactor residues with an exothermic heat content of 123.4 J/g provides the scope for further energy derivation.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: microbiota

Bacterial and Metabolic Character of Traditional Sour Cream and Butter in Buryatia, Russia.

Traditional sour cream and butter are widely popular fermented dairy products in Russia for their flavor and nutrition, and contain rich microbial biodiversity, particularly in terms of lactic bacteria (LAB). However, few studies have described the microbial communities and metabolic character of traditional sour cream and butter. The objective of this study was to determine the bacterial and metabolic character of eight samples collected from herdsmen in Buryatia, Russia. Using single-molecule real-time (SMRT) sequencing techniques, we identified a total of 294 species and/or subspecies in 169 bacterial genera, belonging to 14 phyla. The dominant phylum was Firmicutes (81.47%) and the dominant genus was (59.28%). There were differences between the bacterial compositions of the sour cream and butter samples. The relative abundances of , , and were significantly higher in sour cream than in butter, and the abundance of was significantly lower in sour cream than in butter. Using a pure culture method, 48 strains were isolated and identified to represent seven genera and 15 species and/or subspecies. Among these isolates, subsp. (22.50%) was the dominant LAB species. Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry at elevated energy was used in combination with statistical methods to detect metabolite differences between traditional sour cream and butter. A total of 27,822 metabolites were detected in all samples, and Lys-Lys, isohexanal, , Leu-Val, and 2\'-deoxycytidine were the most dominant metabolites found in all samples. In addition, 27 significantly different metabolites were detected between the sour cream and butter samples, including short peptides, organic acids, and amino acids. Based on correlation analyses between the most prevalent bacterial species and the main metabolites in sour cream, we conclude that there may be a connection between the dominant LAB species and these metabolites. This study combined omics techniques to analyze the bacterial diversity and metabolic character of traditional sour cream and butter, and we hope that our findings will enrich species resource libraries and provide valuable resources for further research on dairy product flavor.

Keyword: microbiota

Biomass and community structure of the abyssal determined from the ester-linked phospholipids recovered from Venezuela Basin and Puerto Rico Trench sediments.

Extractible phospholipid fatty acids of abyssal sediment cores from three stations in the Venezuela Basin, transects between them, and a station in the Puerto Rico Trench were analyzed to determine microbial biomass and community composition. Results were compared to abyssal sediments from an area of high-energy boundary currents in the North Atlantic, and estuarine sediments from Apalachee Bay, Florida. Venezuela Basin and Puerto Rico Trench sediments were characterize by low microbial biomass, measured as phospholipid . Venezuela Basin sediments of three different sedimentary regimes showed a remarkably similar microbial community structure, as characterized by fatty profiles. Prokaryotic organisms dominated the microbial community, and fatty acids believed to be signatures of anaerobic organisms were present in greater proportions in Venezuela Basin and Puerto Rico Trench sediments than in either the North Atlantic abyssal sediments or shallow-water estuarine sediments.

Keyword: microbiota

Utility of high performance liquid chromatography/electrospray/mass spectrometry of polar lipids in specifically Per-13C labeled Gram-negative bacteria DA001 as a tracer for acceleration of bioremediation in the subsurface.

Specific fatty acids from phosphatidylglycerol (PG) and phosphatidylethanolamine (PE) recovered from a per 13C-labeled bacteria can be detected in environmental samples and used as measures of bacterial transport in the subsurface. Detection of (16:0) and oleic (18:1) at m/z 271 (255+16) and 299 (281+18) as negative ions in PG and PE separated by high performance liquid chromatography (HPLC) and detected after up-front collisionally induced dissociation (CID) utilizing electrospray (ES) mass spectrometry (MS) provided sufficient sensitivity and specificity for detection in the presence of the indigenous . Application of tandem mass spectrometry (MS/MS) in the multiple reaction monitoring (MRM) was use to monitor selected transitions. MRM can increase the sensitivity so that polar lipids recovered from cell densities currently at about 10(4) cells/sample can be detected. This technology provides a non-intrusive mechanism for monitoring the distribution of bacteria added to accelerate in situ bioremediation of subsurface sediments.

Keyword: microbiota

Obesity changes the human gut mycobiome.

The human intestine is home to a diverse range of bacterial and fungal species, forming an ecological community that contributes to normal physiology and disease susceptibility. Here, the fungal (mycobiome) in obese and non-obese subjects was characterized using Internal Transcribed Spacer (ITS)-based sequencing. The results demonstrate that obese patients could be discriminated by their specific fungal composition, which also distinguished metabolically "healthy" from "unhealthy" obesity. Clusters according to genus abundance co-segregated with body fatness, fasting triglycerides and HDL-cholesterol. A preliminary link to metabolites such as hexadecanedioic , caproic and N-acetyl-L-glutamic was also found. Mucor racemosus and M. fuscus were the species more represented in non-obese subjects compared to obese counterparts. Interestingly, the decreased relative abundance of the Mucor genus in obese subjects was reversible upon weight loss. Collectively, these findings suggest that manipulation of gut mycobiome communities might be a novel target in the treatment of obesity.

Keyword: microbiota

Antioxidant potential and phenolic profile of blackberry anthocyanin extract followed by human gut fermentation.

Beneficial properties attributed to the intake of blackberry fruit are associated with the presence of high content of anthocyanins. However, their low absorption and accumulation in the gut have generated the belief that gut metabolites of anthocyanins are probably reason for their protective effects. In this study, blackberry anthocyanins were prepared and subjected to in vitro human gut fermentation at different time intervals (0-48\u202fh) to study their gut metabolites and antioxidant properties. The content of cyanidin-3-O-glucoside was found highest in blackberry and it degraded completely after 6\u202fh fermentation. Gut metabolites of blackberry anthocyanins were found to improve the glucose consumption and glycogen content significantly in HepG2 cells. Furthermore, gut metabolites significantly ameliorated high glucose plus (HG\u202f+\u202fPA)-induced ROS, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. Overall, this study reveals that blackberry anthocyanins subjected to gut fermentation resulted in the formation of active metabolites with potential antioxidant activity against HG\u202f+\u202fPA-induced oxidative stress.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiota

Alterations to the -colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. -colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut , colonic inflammation and cognitive function to elucidate the role of -gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut . DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut , colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: microbiota

Structure-function relationship of the milk fat globule.

The complex biochemical composition and physical structure of the milk fat globule (MFG) are presented as a basis for its paradoxical metabolic fate: MFG is a rapid conveyor of energy through its triacylglycerol (TAG) core but contains some low-digestible bioactive complex lipids and proteins, which influence lipid metabolism and contribute to intestinal and systemic health.MFG structure modulates gastrointestinal lipolysis, postprandial lipemia and even the postprandial fate of ingested fatty acids. Proof-of-concept of the nutritional programming induced by early consumption of an emulsion biomimetic of MFG compared with a typical infant formula was published in an animal model (mice). The metabolic response to a high-fat diet during adulthood was improved following neonatal exposure to the biomimetic emulsion.MFG TAG are tailored with a unique regiodistribution delivering in priority short to medium-chain fatty acids in gastric phase, an important amount of quickly metabolizable oleic and protecting in sn-2 position. MFG digestion may not only trigger rapid TAG and chylomicron plasma peaks with fast clearance but also the luminal release of nonhydrolysable bioactive compounds (glycosylated compounds and sphingomyelin), which contribute to intestinal and systemic health by shaping the and modulating the immune system. These bioactive compounds form self-assembled structures, protect specific micronutrients and lower cholesterol absorption. The health benefits of MFG consumption or of some of its fractions (MFGM) under specific structures are steadily being demonstrated with still much unsolved questions especially for populations with high nutritional needs (e.g. elderly, infants).

Keyword: microbiota

Exploring the interactions between serum free fatty acids and fecal in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut is involved in the host energy metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal seems to exist. Our aim was to explore the interaction among serum FFA levels, gut , diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiota

Augmented hepatic Toll-like receptors by fatty acids trigger the pro-inflammatory state of non-alcoholic fatty liver disease in mice.

There is considerable evidence that intestinal are involved in the development of metabolic syndromes and, consequently, with the development of non-alcoholic fatty liver disease (NAFLD). Toll-like receptors (TLRs) are essential for the recognition of . However, the induction mechanism of TLR signals through the gut-liver axis for triggering the development of non-alcoholic steatohepatitis (NASH) or NAFLD remains unclear. In this study, we investigated the role of (PA) in triggering the development of a pro-inflammatory state of NAFLD.Non-alcoholic fatty liver disease was induced in mice fed a high fat diet (HFD). The mice were killed and the expression of TLRs, tumor necrosis factor (TNF), interleukin (IL)-1β, and phospho-interleukin-1 receptor-associated kinase 1 in the liver and small intestine were assessed. In addition, primary hepatocytes and Kupffer cells were treated with PA, and the direct effects of PA on TLRs induction by these cells were evaluated.The expression of inflammatory cytokines such as TNF, IL-1β, and TLR-2, -4, -5, and -9 was increased in the liver, but decreased in the small intestine of HFD-fed mice in vivo. In addition, the expression of TLRs in primary hepatocytes and Kupffer cells was increased by treatment with PA.In the development of the pro-inflammatory state of NAFLD, PA triggers the expression of TLRs, which contribute to the induction of inflammatory cytokines through TLR signals by intestinal .© 2013 The Japan Society of Hepatology.

Keyword: microbiota

Gut and Metabolome Response of Seed Oil on Metabolism Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.

Keyword: microbiota

Effect of high β-palmitate content in infant formula on the intestinal of term infants.

(PA) constitutes 17% to 25% of the human milk fatty acids, and ~70% is esterified in the sn-2 position of triglycerides (β-palmitate). In the sn-2 position, PA is not hydrolyzed and thus is efficiently absorbed. The PA in palm oils, commonly used in infant formulas, is esterified in the sn-1 and sn-3 positions. In these positions, PA is hydrolyzed and forms poorly absorbed calcium complexes. The present study assessed whether high β-palmitate in infant formulas affects the intestinal flora.Thirty-six term infants were enrolled: 14 breast-fed (BF group) and 22 formula-fed infants who were randomly assigned to receive formula containing high β-palmitate (HBP group, n=14), or low β-palmitate (LBP group, n=8), where 44% and 14% of the PA was β-palmitate, respectively. The total amount of PA in the formulas was 19% and 22% in the LBP and HBP groups, respectively. Neither formula contained pre- or probiotics. Stool samples were collected at enrollment and at 6 weeks for the quantification of bacteria.At 6 weeks, the HBP and BF groups had higher Lactobacillus and bifidobacteria counts than the LBP group (P<0.01). The Lactobacillus counts at 6 weeks were not significantly different between the HBP and BF groups. Lactobacillus counts were 1.2×10¹⁰, 1.2×10¹¹, and 5.6×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively. Bifidobacteria counts were 5.1×10⁹, 1.2×10¹¹, and 3.9×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively.HBP formula beneficially affected infant gut by increasing the Lactobacillus and bifidobacteria counts in fecal stools.ClinicalTrials.gov .

Keyword: microbiota

[The oleic triglycerides of palm oil and triglycerides of creamy fat. The reaction of palmitoylation, potassium and magnesium palmitate, absorption of fatty acids by enterocytes and of large intestine].

The decreasing of content of animal, milk fat (butter) by means of its substitution with vegetable, oleic, palmy oil in food of adults optimal by its quantity is physically chemically and biologically substantiated. In oleic palmy oil higher content of oleic mono unsaturated fatty and oleic triglycerides than in creamy fat is established. The biologic availability of unsaturated in the form of free fatty is decreased at its absorption by enterocytes of small intestines is detected. There are no transforms of mono unsaturated acids in palmy oil in contrast with hydrogenated margarines. In palmy, oleic oil there is not enough of short-chained fatty acids (C4-C6) and it has no taste quality and it has low level of unsaturated fatty acids and factually it is lacking of ω-6 polyunsaturated fatty acids. However, it is compensated in case of availability offish and sea products in food. If adults, especially older ones, will refuse to consume creamy fat and decrease intake of products with high content of unsaturated fatty and triglycerides (beef, sour cream, fatty cheeses) it\'ll positively impact their health. The refusal from these products is a real step in prevention of metabolic pandemic (atherosclerosis and atheromatosis, metabolic syndrome, resistance to insulin, obesity). There are still large number of people who at optimal amount of food retain in vivo increased amount of exogenous, endogenously synthesized from glucose unsaturated fatty in the form of unesterified fatty acids (syndrome of resistance to insulin) and increased content of triglycerides.

Keyword: microbiota

Rhizosphere effect of Scirpus triqueter on soil microbial structure during phytoremediation of diesel-contaminated wetland.

Though phytoremediation has been widely used to restore various contaminated sites, it is still unclear how soil microbial communities respond microecologically to plants and pollutants during the process. In this paper, batch microcosms imitating in situ phytoremediation of petroleum-contaminated wetland by Scirpus triqueter were set up to monitor the influence of plant rhizosphere effect on soil microbes. , one of the main root exudates of S. triqueter, was added to strengthen rhizosphere effect. Abundances of certain microbial subgroups were quantified by phospholipid fatty profiles. Results showed that diesel removal extents were significantly higher in the rhizosphere (57.6 +/-4.2-65.5 +/- 6.9%) than those in bulk soil (27.8 +/-6.5-36.3 +/- 3.2%). In addition, abundances of saturated, monounsaturated, and polyunsaturated fatty acids were significantly higher (P < 0.05) in planted soil than those in the bulk soil. When it was less than 15,000 mg diesel kg soil-1, increasing diesel concentration led to higher abundances of fungi, Gram-positive and Gram-negative bacteria. The addition of amplified the rhizosphere effect on soil microbial populations and diesel removal. Principal component analysis revealed that plant rhizosphere effect was the dominant factor affecting microbial structure. These results provided new insights into plant-microbe-pollutant coactions responsible for diesel degradation, and they were valuable to facilitate phytoremediation of diesel contamination in wetland habitats.

Keyword: microbiota

Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut , and hepatic metabolism.

Keyword: microbiota

Regulation of -GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of -GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: microbiota

Effect of silicate grain shape, structure, and location on the biomass and community structure of colonizing marine .

colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable . With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined.

Keyword: microbiota

Tuning culturing conditions towards the production of neutral lipids from lubricant-based wastewater in open mixed bacterial communities.

Production of bacterial lipid-based biofuels using inexpensive substrates, as wastes, is an emerging approach. In this work, a selective process using carbon feast-famine cycles was applied to obtain an indigenous microbial community of hydrocarbon-degrading and lipid-accumulating bacteria, using a real lubricant-based wastewater as carbon source. In the conditions applied, the enriched bacterial community, dominated by members of the genus Rhodococcus, Pseudomonas and Acinetobacter, was able to degrade almost all hydrocarbons present in the wastewater within 24\u202fh\' incubation and to accumulate, although in low levels, triacylglycerol (TAG) (<5% of cell dry weight (CDW)) and polyhydroxyalkanoates (PHA) (3.8%\u202f±\u202f1.1% of the CDW) as well as an unknown lipid (29%\u202f±\u202f6% of CDW), presumably a wax ester-like compound. The influence of culture conditions, namely carbon and nitrogen concentrations (and C/N ratio) and cultivation time, on the amount and profile of produced storage compounds was further assessed using a statistical approach based on a central composite circumscribed design and surface response methodology. The regression analysis of the experimental design revealed that only nitrogen concentration and C/N ratio are significant for neutral lipid biosynthesis (p\u202f<\u202f0.05). Maximum neutral lipid content, i.e. 33% (CDW basis), was achieved for the lowest carbon and nitrogen concentrations evaluated (10\u202fg COD L and 0.02\u202fg\u202fN L). PHA accounted for less than 5% of CDW. In these conditions, neutral lipid content was mainly composed by TAG, about 70% (w/w). TAG precursors, namely monoacylglycerols (MAG), diacylglycerols (DAG) and fatty acids (FA), accounted for 22% of total neutral lipids and WE for about 7%. Nevertheless, according to the applied response surface model, further improvement of neutral lipids content is still possible if even lower nitrogen concentrations are used. The fatty acids detected in TAG extracts ranged from myristic (C14:0) to linoleic (C18:2), being the most abundant (C16:0), stearic (C18:0) and oleic (C18:1). This study shows the feasibility of combining treatment of hydrocarbon contaminated wastewater, herein demonstrated for lubricant-based wastewater, with the production of bacterial neutral lipids using open mixed bacterial communities. This approach can decrease the costs associated to both processes and contribute to a more sustainable waste management and production of lipid-based biofuels.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: microbiota

Association between sn-2 fatty profiles of breast milk and development of the infant intestinal microbiome.

Increasing evidence shows that host diet and gut microbes are related. Previous studies have shown the effects of specific dietary fatty acids (FAs) on intestinal , but little is known about the effect of the stereospecifically numbered sn-2 position in triglycerides (TG) of human milk on the gut microbiome of infants. This study aimed at examining possible effects of sn-2 FAs of human milk on the gut microbial development of breastfeeding babies. Sn-2 FAs and intestinal were assessed by GC-MS and high-throughput 16S rRNA sequencing, respectively. The results showed that breast milk from mothers in China contained ten major sn-2 FAs dominated by (C, 54.42%), oleic (C n-9, 14.95%), linoleic (LA, C n-6, 12.81%), myristic (C, 4.50%) and C (3.17%). Total long chain unsaturated fatty acids (LCUFA) decreased from colostrum to mature milk, while total saturated fatty acids (SFA) showed no significant difference during lactation. A significant association between sn-2 FAs in milk and infant gut was found between decanoic (C), myristic (C), stearic (C), C, arachidonic (AA, C n-6), docosahexaenoic (DHA, C n-3) with Bacteroides, Enterobacteriaceae, Veillonella, Streptococcus, and Clostridium. These microbes were involved in short-chain fatty (SCFA) production and other functions, and significantly increased at 13-15 d after breastfeeding was initiated. C and DHA were relevant to most of the microbes. This study demonstrated the relatively steady profiles of sn-2 FAs in breast milk and gut of infants, together with their correlation during the breastfeeding period. The above results provided important information for designing the configuration of FAs in next-generation formulas for Chinese infants.

Keyword: microbiota

Metabolic pathway and role of individual species in the bacterial consortium for biodegradation of azo dye: A biocalorimetric investigation.

In this study, an attempt was made to investigate the functional role and metabolic behaviour of the monoculture (Staphylococcus lentus (SL), Bacillus flexus (BF) and Pseudomonas aeruginosa (PA)) in the bacterial biocenosis for biotransformation of an azo dye. The power-time profile obtained from consortia depicted three distinct peaks, which correlated well with the individual bacterial growth (PA\xa0>\xa0SL\xa0>\xa0BF), indicating the synergistic relation and division of labour in the biocenosis. The heat release pattern was used to identify the sequential behaviour of microbial consortia in real time. Yield calculation based on total heat liberated to the complete substrate utilization Y for PA, SL, and BF were 15.99, 16.68, 7.32\xa0kJ/L respectively. Similarly, the oxy calorific values Y for the above species are respectively 386, 375, 440\xa0kJ/mol and indicates the aerobic nature of microorganism employed. Further, the metabolome produced during the biotransformation were identified using Gas Chromatography-Mass Spectrometry (GC-MS), based on which a plausible pathway was predicted. The abundant metabolites were (m/z\xa0=\xa0256) and diethyl phthalate (m/z\xa0=\xa0222.2). The abundance of diethyl phthalate was much lesser in the consortia compared to the monoculture. Thus, the biocalorimetric heat yield calculation along with the stoichiometry and plausible pathway based biochemical elucidation provides a mechanistic basis for understanding the azo-dye biotransformation by the monocultures in consortia.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: microbiota

Mode of action of a fatty -based natural product to control Botrytis cinerea in grapes.

To investigate the efficacy and mode of action of the fatty -based product Foodcoat(®) (FC) against Botrytis cinerea.In vitro, in vivo and field experiments were carried out to investigate the effect of different concentrations of FC on B. cinerea germination and infection of grape leaves and berries, using three selected isolates and comparing results with those achieved by the commercialized product Protector(HML) (PRT). Furthermore, the effect of field applications of FC on the grape berry was investigated. FC reduced B. cinerea germination and grape berry severity by up to 54 and 96%, respectively, compared with the untreated controls.Foodcoat demonstrated efficacy that was equal or greater than the registered product, PRT. A multiple mode of action was hypothesized for FC suppression of B. cinerea, including: inhibition of germination and germ tube alteration, protection of host green tissues and enhancement of the natural yeast populations on the berry surface.The efficacy of both products has been quantified and their modes of action described, suggesting them for field applications against B. cinerea, alone or in combined strategies. This is also the first report of a fatty -based product stimulating natural yeast populations on grape berries.© 2014 The Society for Applied Microbiology.

Keyword: microbiota

Characterization of aerobic oil and grease-degrading bacteria in wastewater.

A bacterial consortium that degrades cooking oil (CO) has been isolated in wastewater (WW) samples, by enrichment in olive CO. This consortium could degrade 90% of CO within 7-9 days (from an initial 1% [w/v]), and it is more active at alkaline conditions. The 16S ribonucleic (RNA) gene analysis showed that it contains five bacterium species: Stenotrophomonas rhizophila, Sphingobacterium sp., Pseudomonas libanensis, Pseudomonas poae and Pseudomonas aeruginosa. This consortium can degrade the free fatty acids (FFA): , stearic, oleic, linoleic and linolenic acids; glycerol, glucose and amylose; and albumin, but could not efficiently degrade carboxymethyl-cellulose. Each strain could also degrade CO and FFAs. The level of bacterial crude-activity of extracellular lipases was found to be between 0.2 and 4U/ml. Using synthetic WW, the consortium could reduce 80% of the chemical oxygen demand [from 10550\u2009±\u20092828\u2005mg/l], 80% of nitrogen (from 410\u2009±\u200978\u2009mgl/l) and 57% of phosphorus (from 93\u2009±\u200925\u2005mg/l). Thus, this consortium can be utilized in the removal of CO from WW.

Keyword: microbiota

Gut -mediated generation of saturated fatty acids elicits inflammation in the liver in murine high-fat diet-induced steatohepatitis.

The gut plays crucial roles in the development of non-alcoholic steatohepatitis (NASH). However, the precise mechanisms by which alterations of the gut and its metabolism contributing to the pathogenesis of NASH are not yet fully elucidated.Mice were fed with a recently reported new class of high-fat diet (HFD), steatohepatitis-inducing HFD (STHD)-01 for 9\xa0weeks. The composition of the gut was analyzed by T-RFLP. Luminal metabolome was analyzed using capillary electrophoresis and liquid chromatography time-of-flight mass spectrometry (CE- and LC-TOFMS).Mice fed the STHD-01 developed NASH-like pathology within a short period. Treatment with antibiotics prevented the development of NASH by STHD-01. The composition of the gut and its metabolic activities were markedly perturbed in the STHD-01-fed mice, and antibiotic administration normalized these changes. We identified that long-chain saturated fatty and n-6 fatty metabolic pathways were significantly altered by STHD-01. Of note, the changes in gut lipidome caused by STHD-01 were mediated by gut , as the depletion of the gut could reverse the perturbation of these metabolic pathways. A saturated long-chain fatty , , which accumulated in the STHD-01 group, activated liver macrophages and promoted TNF-α expression.Lipid metabolism by the gut , particularly the saturation of fatty acids, affects fat accumulation in the liver and subsequent liver inflammation in NASH.

Keyword: microbiota

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile Metabolism and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut , is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) metabolism. However, whether TMAO aggravates liver steatosis by modulating BA metabolism and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA metabolism. Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: microbiota

Beta-palmitate - a natural component of human milk in supplemental milk formulas.

The composition and function of human milk is unique and gives a basis for the development of modern artificial milk formulas that can provide an appropriate substitute for non-breastfed infants. Although human milk is not fully substitutable, modern milk formulas are attempting to mimic human milk and partially substitute its complex biological positive effects on infants. Besides the immunomodulatory factors from human milk, research has been focused on the composition and structure of human milk fat with a high content of β- (sn-2 , β-palmitate). According to the available studies, increasing the content of β-palmitate added to milk formulas promotes several beneficial physiological functions. β-palmitate positively influences fatty metabolism, increases calcium absorption, improves bone matrix quality and the stool consistency, and has a positive effect on the development of the intestinal microbiome.

Keyword: microbiota

Characterisation of Fecal Soap Fatty Acids, Calcium Contents, Bacterial Community and Short-Chain Fatty Acids in Sprague Dawley Rats Fed with Different sn-2 Triacylglycerols Diets.

The structure of dietary triacylglycerols is thought to influence fatty and calcium absorption, as well as intestinal population of the host. In the present study, we investigated the impact of (PA) esterified at the sn-2 position on absorption of fatty and calcium and composition of intestinal microorganisms in rats fed high-fat diets containing either low sn-2 PA (12.1%), medium sn-2 PA (40.4%) or high sn-2 PA (56.3%), respectively. Fecal fatty profiles in the soaps were measured by gas chromatography (GC), while fecal calcium concentration was detected by ICP-MS. The fecal microbial composition was assessed using a 16S rRNA high-throughput sequencing technology and fecal short-chain fatty acids were detected by ion chromatograph. Dietary supplementation with a high sn-2 PA fat significantly reduced total fecal contents of fatty acids soap and calcium compared with the medium or low sn-2 PA fat groups. Diet supplementation with sn-2 PA fat did not change the entire profile of the gut community at phylum level and the difference at genera level also were minimal in the three treatment groups. However, high sn-2 PA fat diet could potentially improve total short-chain fatty acids content in the feces, suggesting that high dietary sn-2 PA fat might have a beneficial effect on host intestinal health.

Keyword: microbiota

Lipid Metabolic Versatility in spp. Yeasts Studied through Metabolic Modeling.

species are lipophilic and lipid-dependent yeasts belonging to the human and animal . Typically, they are isolated from regions rich in sebaceous glands. They have been associated with dermatological diseases such as seborrheic dermatitis, pityriasis versicolor, atopic dermatitis, and folliculitis. The genomes of , , and lack the genes related to fatty synthesis. Here, the lipid-synthesis pathways of these species, as well as of , and of an atypical variant were reconstructed using genome data and Constraints Based Reconstruction and Analysis. To this end, the genomes of CBS 1878 and the atypical 4DS were sequenced and annotated. The resulting Enzyme Commission numbers and predicted reactions were similar to the other strains despite the differences in their genome size. Proteomic profiling was utilized to validate flux distributions. Flux differences were observed in the production of steroids in and in the metabolism of butanoate in . The predictions obtained via these metabolic reconstructions also suggested defects in the assimilation of in , , , and the atypical variant of , but not in These predictions were validated via physiological characterization, showing the predictive power of metabolic network reconstructions to provide new clues about the metabolic versatility of .

Keyword: microbiota

Dynamic alterations in the gut and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut and metabolome.

Keyword: microbiota

Lipotoxicity and the gut-liver axis in NASH pathogenesis.

The pathogenesis of non-alcoholic fatty liver disease, particularly the mechanisms whereby a minority of patients develop a more severe phenotype characterised by hepatocellular damage, inflammation, and fibrosis is still incompletely understood. Herein, we discuss two pivotal aspects of the pathogenesis of NASH. We first analyse the initial mechanisms responsible for hepatocellular damage and inflammation, which derive from the toxic effects of excess lipids. Accumulating data indicate that the total amount of triglycerides stored in hepatocytes is not the major determinant of lipotoxicity, and that specific lipid classes act as damaging agents on liver cells. In particular, the role of free fatty acids such as , cholesterol, lysophosphatidylcholine and ceramides has recently emerged. These lipotoxic agents affect the cell behaviour via multiple mechanisms, including activation of signalling cascades and death receptors, endoplasmic reticulum stress, modification of mitochondrial function, and oxidative stress. In the second part of this review, the cellular and molecular players involved in the cross-talk between the gut and the liver are considered. These include modifications to the , which provide signals through the intestine and bacterial products, as well as hormones produced in the bowel that affect metabolism at different levels including the liver. Finally, the activation of nuclear receptors by bile acids is analysed.Copyright © 2017. Published by Elsevier B.V.

Keyword: microbiota

The influence of the position of palmitate in infant formula triacylglycerols on health outcomes.

The purpose of this review is to discuss recent studies reporting on the influence of the position of in triacylglycerols in infant formula and relevant animal studies. Earlier experiments in rodents show that a diet with a higher proportion of palmitate at the sn-2 position of triacylglycerols improves dietary fat and calcium absorption compared with a diet with a lower sn-2 palmitate content. A high-sn-2 palmitate diet increased fecal short-chain fatty acids, reduced gut inflammation in a colitis model, and altered tissue endocannabinoid concentrations in laboratory rodents. Recent studies in infants confirm that formula with a high sn-2 palmitate content reduces stool fat, , fat soaps, palmitate soaps, and calcium compared with formula with a low sn-2 palmitate content. These effects have been associated with improved bone strength, increased fecal bifidobacteria, and reduced crying in infants. In some studies, findings with formula high in sn-2 palmitate match those seen in breast-fed infants. However, in many studies, high sn-2 palmitate formula remains inferior to breast-feeding. It is concluded that infant formula high in sn-2 palmitate is superior to formula with low sn-2 palmitate but does not fully match human breast milk. Recent studies showing altered gut (human infants) and tissue endocannabinoids (rodent model) suggest the potential for marked physiological impact of high sn-2 palmitate that needs to be explored further in human trials.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: microbiota

Time-Dependent alteration to the tight junction structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal composition, increased intestinal permeability, and onset of type 2 diabetes mellitus (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the tight junction (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the , after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: microbiota

Anaerococcus rubiinfantis sp. nov., isolated from the gut of a Senegalese infant with severe acute malnutrition.

Anaerococcus rubiinfantis sp. nov. strain mt16(T) is a new species within the genus Anaerococcus, which was isolated by the culturomics approach from the gut of an infant suffering from kwashiorkor. A phenotypic, biochemical and proteomic description of this strain is hereby presented alongside a complete annotation of its genome. This strictly anaerobic species forms Gram-positive non-sporeforming cocci. The major fatty was hexadecanoic . The phylogenetic analysis of strain mt16(T) showed a 97.9% similarity level with Anaerococcus vaginalis, the closest validly published species. Its genome is 1,929,161\xa0bp long with 29.5% G\xa0+\xa0C content and contains 1808 protein-coding genes and 56 RNA genes, among which are six rRNA genes. Genomic analysis identified 41/1864 coding genes as ORFans (2.2%) and at least 620/1808 (34.9%) orthologous proteins which are not shared with the closest phylogenetic species. We believe that the extension of the human anaerobic gut compendium by culturomics is one of the first steps that will improve the understanding of the links between the microbiome and health or disease.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: microbiota

D-chiro-inositol enriched Fagopyrum tataricum (L.) Gaench extract alleviates mitochondrial malfunction and inhibits ER stress/JNK associated inflammation in the endothelium.

Tartary buckwheat is a food medicine dual-use crop with healing effects on cardiovascular diseases and type2 diabetes. It has been proposed that endothelial dysfunction is the initial lesion in these diseases and it\'s associated with mitochondrial dysfunction, endoplasmic reticulum (ER) stress and inflammation. D-chiro-inositol (DCI) is a bioactive compound of Tartary buckwheat and is always deficit in type2 diabetes. However, it remains unknown whether DCI-enriched Tartary buckwheat extract can ameliorate mitochondrial dysfunction, ER stress and inflammation in the endothelium.Endothelial cells were treated with (PA) and mice were fed with high fat diet (HFD). The effects of DCI-enriched Tartary buckwheat bran extract (TBBE) on superoxide anion generation, dynamin-related protein 1 (Drp1), mitofusin2 (Mfn2), inositol-requiring enzyme-1α (IRE1α) and Jun n-terminal kinase (JNK) activation and inflammation in the endothelium against lipotoxicity were investigated.In endothelial cells, TBBE significantly inhibited oxidative stress. Meanwhile, in HFD-fed mice and PA-induced cells, TBBE regulated Drp1 phosphorylation and inhibited its activation, implying the protective effect of TBBE on mitochondrial morphology. As a result, TBBE protected mitochondrial function. Additionally, TBBE inhibited ER stress and reduced the production of IL-6 and VCAM-1, associated with JNK pathway, thereby inhibiting the caspase-3 activation in vivo and in vitro.Taken together, this study indicated the beneficial role of TBBE in endothelial inflammation, with emphasis on mitochondrial dysfunction, ER stress and JNK activation.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Involvement of palmitate/Ca2+(Sr2+)-induced pore in the cycling of ions across the mitochondrial membrane.

The palmitate/Ca2+-induced (Pal/Ca2+) pore, which is formed due to the unique feature of long-chain saturated fatty acids to bind Ca2+ with high affinity, has been shown to play an important role in the physiology of . The present study demonstrates that the efflux of Ca2+ from rat liver induced by ruthenium red, an inhibitor of the energy-dependent Ca2+ influx, seems to be partly due to the opening of Pal/Ca2+ pores. Exogenous Pal stimulates the efflux. Measurements of pH showed that the Ca2+-induced alkalization of the mitochondrial matrix increased in the presence of Pal. The influx of Ca2+ (Sr2+) also induced an outflow of K+ followed by the reuptake of the ion by . The outflow was not affected by a K+/H+ exchange blocker, and the reuptake was prevented by an ATP-dependent K+ channel inhibitor. It was also shown that the addition of Sr2+ to under hypotonic conditions was accompanied by reversible cyclic changes in the membrane potential, the concentrations of Sr2+ and K+ and the respiratory rate. The cyclic changes were effectively suppressed by the inhibitors of Ca2+-dependent phospholipase A2, and a new Sr2+ cycle could only be initiated after the previous cycle was finished, indicating a refractory period in the mitochondrial sensitivity to Sr2+. All of the Ca2+- and Sr2+-induced effects were observed in the presence of cyclosporin A. This paper discusses a possible role of Pal/Ca2+ pores in the maintenance of cell ion homeostasis.Copyright © 2014 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Cytochrome c release from rat liver is compromised by increased saturated cardiolipin species induced by sucrose feeding.

Cytochrome c release from has been described to be related to reactive oxygen species (ROS) generation. With ROS generation being increased in fatty liver from sucrose-fed (SF) rats, we hypothesized that cytochrome c release might be positively associated with H2O2 generation from SF . Surprisingly, cytochrome c release from of SF liver was found to be significantly lower compared with control (C) oxidizing pyruvate/malate or succinate. Exposure of to exogenous superoxide radical generated by the xanthine/xanthine oxidase system elicits a dose-response cytochrome c release in both control and SF , but cytochrome c release remains lower in SF compared with C . Furthermore, the addition of ebselen, PEG-catalase, or catalase, a H2O2 scavenger, significantly reduces cytochrome c release from C and SF . Our results suggest that both intra- and extramitochondrial H2O2 are involved in cytochrome c release, but the persisting difference between C and SF levels can be attributed to the differences in cardiolipin compositions. Indeed, the ratio of -rich cardiolipin species was found to be increased in lipid membrane from SF compared with C , whereas that of linoleic -rich cardiolipin species was found decreased. In addition, the content of tafazzin, a protein responsible for cardiolipin remodeling, was decreased in SF . Therefore, we conclude that the changes observed in the composition of cardiolipin molecular species in SF may be involved in cytochrome c interaction with mitochondrial inner membrane lipid and in its reduced release from SF .Copyright © 2015 the American Physiological Society.

Keyword: mitochondria

Urinary metabolomics for discovering metabolic biomarkers of laryngeal cancer using UPLC-QTOF/MS.

The incidence of laryngeal cancer (LYC) is second only to lung cancer, which is also the second most common cancer in head and neck cancer. Risk assessment metabolomics biomarkers to diagnose LYC have not been found by now. To profile the metabolites in healthy controls (HCs) and LYC patients (LYCs), urine metabonomics study was performed based on reversed phase liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RPLC-QTOF/MS). Six urine differentially expressed metabolites (Variable Importance in Projection >1, p\u2009<\u2009 0.05) were identified by searching reference library or comparing with standard based on OPLS-DA (orthogonal partial least squares-discriminant analysis) model. d-pantothenic , , myristic , oleamide, sphinganine and phytosphingosine were identified as differential metabolites associated with the LYC and they might play roles in sphingolipid metabolism, fatty biosynthesis, fatty elongation in , pantothenate and coenzyme A (CoA) biosynthesis, beta-Alanine metabolism and fatty metabolism. These six differential metabolites were combined to test the potentiality of diagnosis of LYC. Results revealed that the area under the curve (AUC) value, sensitivity and specificity of receiving operator characteristic (ROC) curve were 0.97, 95% and 97%, respectively, indicating that this diagnosis method could be used to distinguish LYCs from HCs with good sensitivity and specificity.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Overexpression of Acyl-ACP Thioesterases, and , Induce Distinct Gene Expression Reprogramming in Developing Seeds of .

We examined the substrate preference of acyl-ACP thioesterases, and , and gene expression changes associated with the modification of lipid composition in the seed, using transgenic plants overexpressing or under the control of a seed-specific promoter. seeds contained a higher level of total saturated fatty (FA) content, with 4.3 times increase in 16:0 , whereas seeds showed approximately 3% accumulation of 10:0 and 12:0 medium-chain FAs, and a small increase in other saturated FAs, resulting in higher levels of total saturated FAs. RNA-Seq analysis using entire developing pods at 8, 25, and 45 days after flowering (DAF) showed up-regulation of genes for β-ketoacyl-acyl carrier protein synthase I/II, stearoyl-ACP desaturase, oleate desaturase, and linoleate desaturase, which could increase unsaturated FAs and possibly compensate for the increase in 16:0 at 45 DAF in transgenic plants. In transgenic plants, many putative chloroplast- or mitochondria-encoded genes were identified as differentially expressed. Our results report comprehensive gene expression changes induced by alterations of seed FA composition and reveal potential targets for further genetic modifications.

Keyword: mitochondria

-Induced Podocyte Apoptosis via the Reactive Oxygen Species-Dependent Mitochondrial Pathway.

Chronic kidney disease (CKD) is often accompanied by hyperlipidemia, which accelerates progression of the disease. Podocyte injury can lead to dysfunction of the glomerular filtration barrier, which is associated with proteinuria, a risk marker for the progression of CKD. Our previous studies demonstrated that (PA) can induce podocyte apoptosis; however, the underlying mechanisms are unclear. In the present study, we investigated the specific molecular mechanisms of PA-induced apoptosis in cultured podocytes.We cultured mouse podocytes and treated them with PA. Then, cell viability was measured using the Cell Counting Kit-8 colorimetric assay, lipid uptake was assessed by Oil Red O staining and boron-dipyrromethene staining, apoptosis was measured by flow cytometry, mitochondrial injury was assessed by JC-1 staining and transmission electron microscopy, and mitochondrial production of reactive oxygen species (ROS) was evaluated by fluorescence microscopy using the MitoSOX Red reagent. The effects of PA on the -mediated caspase activation pathway were investigated by examining the expression of caspase-8, cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), B-cell lymphoma 2 (Bcl-2), Bax, Bid, cytochrome c, and Fas-associated protein with death domain (FADD) using western blotting. The translocation of Bax and cytochrome c were detected by immunofluorescence.PA treatment significantly increased lipid accumulation and induced podocyte apoptosis. We investigated whether the two primary apoptosis signaling pathways (death receptor-mediated pathway and -mediated pathway) were involved in the execution of PA-induced podocyte apoptosis, and found that the levels of FADD, caspase-8, and Bid did not significantly change during this process. Meanwhile, PA treatment induced an increase in Bax protein expression and a decrease in Bcl-2 protein expression, with Bax translocation to the . Furthermore, PA treatment induced mitochondrial impairment, and triggered the release of cytochrome c from the to cytosol, with a concomitant dose-dependent increase in the levels of cleaved caspase-9, cleaved caspase-3, and PARP. Meanwhile, PA treatment increased mitochondrial production of ROS, and the -targeted antioxidant mitoTEMPO significantly ameliorated PA-induced podocyte apoptosis.Our findings indicated that PA induced caspase-dependent podocyte apoptosis through the mitochondrial pathway, and mitochondrial ROS production participated in this process, thus potentially contributing to podocyte injury.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: mitochondria

Membranotropic effects of ω-hydroxypalmitic and Ca on rat liver and lecithin liposomes. Aggregation and membrane permeabilization.

The paper examines membranotropic Ca-dependent effects of ω-hydroxypalmitic (HPA), a product of ω-oxidation of fatty acids, on the isolated rat liver and artificial membrane systems (liposomes). It was established that in the presence of Ca, HPA induced aggregation of liver , which was accompanied by the release of cytochrome c from the organelles. It was further demonstrated that the addition of Ca to HPA-containing liposomes induced their aggregation and/or fusion. Ca also caused the release of the fluorescent dye sulforhodamine B from liposomes, indicating their permeabilization. HPA was shown to induce a high-amplitude swelling of Ca-loaded , to decrease their membrane potential, to induce the release of Ca from the organelles and to result in the oxidation of the mitochondrial NAD(P)H pool. Those effects of HPA were not blocked by the MPT pore inhibitor CsA, but were suppressed by the mitochondrial calcium uniporter inhibitor ruthenium red. The effects of HPA were also observed when Ca was replaced with Sr (but not with Ba or Mg). A supposition is made that HPA can induce a Ca-dependent aggregation of , as well as Cadependent CsA-insensitive permeabilization of the inner mitochondrial membrane - with the subsequent lysis of the organelles.

Keyword: mitochondria

Dual roles of ULK1 (unc-51 like autophagy activating kinase 1) in cytoprotection against lipotoxicity.

Saturated fatty (SFA)-induced lipotoxicity is caused by the accumulation of reactive oxygen species (ROS), which is associated with damaged . Moreover, lipotoxicity is crucial for the progression of nonalcoholic steatohepatitis (NASH). Autophagy is required for the clearance of protein aggregates or damaged to maintain cellular metabolic homeostasis. The NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2)-KEAP1 (kelch like ECH associated protein 1) pathway is essential for the elimination of ROS. ULK1 (unc-51 like autophagy activating kinase 1; yeast Atg1) is involved in the initiation of autophagy; however, its role in lipotoxicity-induced cell death in hepatocytes and mouse liver has not been elucidated. We now show that ULK1 potentiates the interaction between KEAP1 and the autophagy adaptor protein SQSTM1/p62, thereby mediating NFE2L2 activation in a manner requiring SQSTM1-dependent autophagic KEAP1 degradation. Furthermore, ULK1 is required for the autophagic removal of damaged and to enhance binding between SQSTM1 and PINK1 (PTEN induced kinase 1). This study demonstrates the molecular mechanisms underlying the cytoprotective role of ULK1 against lipotoxicity. Thus, ULK1 could represent a potential therapeutic target for the treatment of NASH. Abbreviations: ACTB: actin beta; CM-HDCFDA:5-(and-6)-chloromethyl-2\',7\'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; LPS: lipopolysaccharides; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAPK8/JNK: mitogen-activated protein kinase 8; MEF: mouse embryonic fibroblast; MFN1: mitofusin 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NASH: nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: ; PARP: poly (ADP-ribose) polymerase 1; PINK1: PTEN induced kinase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PRKC/PKC: protein kinase C; RBX1: ring-box 1; ROS: reactive oxygen species; SFA: saturated fatty ; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; TUBA: tubulin alpha; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase 1.

Keyword: mitochondria

Cytotoxic effect of Kalanchoe flammea and induction of intrinsic mitochondrial apoptotic signaling in prostate cancer cells.

Kalanchoe flammea Stapf (Crassulaceae) is a medicinal plant grown in the South of Mexico (State of Tabasco), which is commonly used in traditional medicine for the treatment of fever, wounds, inflammation, and cancer.To establish the potential of K. flammea for the treatment of prostate cancer, evaluating its cytotoxic activity, its probable mechanism of action, and carrying out some toxicological safety studies.The cytotoxic activity of the ethyl acetate extract of K. flammea (Kf-EtOAc) was evaluated in several cell lines of prostate cancer by MTT viability assay. The cellular death mechanism was studied by evaluating the translocation of phosphatidylserine (Annexin V); overproduction of reactive oxygen species [2\'-7\'-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay]; release of Cytochrome C; activation of caspase-3 and\xa0-9, and regulation of Bcl-2, XIAP, and PKCε proteins by Western Blot analysis. For the evaluation of the safety of Kf-EtOAc, the Ames test, Micronucleus assay, and acute toxicity study were determined.Kf-EtOAc exhibited selective cytotoxic activity against prostate cell lines as follows: PC-3, LNCaP, and PrEC (IC = 1.36\u202f±\u202f0.05; 2.06\u202f±\u202f0.02, and 127.05\u202f±\u202f0.07\u202fμg/mL, respectively). The F82-P2 fraction (rich in coumaric and ) obtained by bioassay-guided fractionation of Kf-EtOAc also demonstrated selective cytotoxic activity against PC-3 cells (IC = 1.05\u202f±\u202f0.06\u202fμg/mL). Kf-EtOAc induces apoptosis by the intrinsic pathway; this mechanism of cell death was confirmed after observing that the extract produces phosphatidylserine translocation, overproduction of reactive oxygen species, release of Cytochrome C at mitochondrial level, and activation of caspase-3 and -9. It was also observed that Kf-EtOAc produces significant downregulation of apoptosis-related proteins Bcl-2, XIAP, and PKCε and induces DNA fragmentation and cell cycle arrest. In addition, Kf-EtOAc is non-genotoxic in vitro by Ames test and non-genotoxic in vivo by Micronucleus assay, and no signs of toxicity or death were reported after the administration of a single acute exposure of 2000\u202fmg/kg.K. flammea is a potential candidate for the development of new drugs for the treatment of prostate cancer. However, to propose their use in clinical trials, additional studies are required to understand their pharmacokinetic behavior, as well as the development of a suitable pharmaceutical form.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: mitochondria

The -Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive oxygen species (ROS) production during glucose metabolism in β cells can be exacerbated under hyperglycaemic conditions such as type 2 diabetes (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: mitochondria

Hydrogen Sulphide Treatment Increases Insulin Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells.

Studies have reported attenuation of insulin resistance (IR) by improving phosphorylation of the insulin signalling pathway. However, the upstream molecular signalling pathway is still elusive. In this study, Western blot was used to evaluate the phosphorylation level of the insulin signalling pathway and the AMPK pathway. 2-NBDG was used to evaluate glucose uptake. Ca imaging was used to assess change of intracellular Ca concentration. We found that NaHS enhanced the intracellular Ca concentration and glucose uptake and activated the insulin signalling cascade in a (PA)-induced IR model in C2C12 cells. Furthermore, activation of the IRS1/PI3K/AKT pathway and glucose uptake were decreased when AMPK or CaMKKβ was inhibited. Our study also showed that the mitochondrial electron transport chain, ATP production, and intramitochondrial cAMP declined in the IR model but that this effect was reversed by NaHS, an effect that may be mediated by the Ca/CaMKK2/AMPK and PI3K/AKT pathways. Our data indicate that HS improves activation of the insulin signalling cascade and glucose uptake via activation of the Ca/CaMKK2/AMPK pathway and mitochondrial metabolism in C2C12 cells. Furthermore, NaHS protects mitochondrial function and maintains normal ATP production by activating the cAMP system and the Ca/CaMKK2/AMPK and PI3K/ATK pathways.

Keyword: mitochondria

Saturated lipids decrease mitofusin 2 leading to endoplasmic reticulum stress activation and insulin resistance in hypothalamic cells.

Endoplasmic reticulum (ER) and dysfunction contribute to insulin resistance generation during obesity and diabetes. ER and interact through Mitofusin 2 (MTF2), which anchors in the outer mitochondrial and ER membranes regulating energy metabolism. Ablation of MTF2 leads to ER stress activation and insulin resistance. Here we determine whether lipotoxic insult induced by saturated lipids decreases MTF2 expression leading to ER stress response in hypothalamus and its effects on insulin sensitivity using in vitro and in vivo models. We found that lipotoxic stimulation induced by , but not the monounsaturated palmitoleic , decreases MTF2 protein levels in hypothalamic mHypoA-CLU192 cells. Also, incubation activates ER stress response evidenced by increase in the protein levels of GRP78/BIP marker at later stage than MTF2 downregulation. Additionally, we found that MTF2 alterations induced by , but not palmitoleic, stimulation exacerbate insulin resistance in hypothalamic cells. Insulin resistance induced by is prevented by pre-incubation of the anti-inflammatory and the ER stress release reagents, sodium salicylate and 4 phenylbutirate, respectively. Finally, we demonstrated that lipotoxic insult induced by high fat feeding to mice decreases MTF2 proteins levels in arcuate nucleus of hypothalamus. Our data indicate that saturated lipids modulate MTF2 expression in hypothalamus coordinating the ER stress response and the susceptibility to insulin resistance.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Palmitate is not an effective fuel for pancreatic islets and amplifies insulin secretion independent of calcium release from endoplasmic reticulum.

The aim of the study was to determine the acute contribution of fuel oxidation in mediating the increase in insulin secretion rate (ISR) in response to fatty acids. Measures of mitochondrial metabolism, as reflected by oxygen consumption rate (OCR) and cytochrome c reduction, calcium signaling, and ISR by rat islets were used to evaluate processes stimulated by acute exposure to (PA). The contribution of mitochondrial oxidation of PA was determined in the presence and absence of a blocker of mitochondrial transport of fatty acids (etomoxir) at different glucose concentrations. Subsequent to increasing glucose from 3 to 20 mM, PA caused small increases in OCR and cytosolic calcium (about 20% of the effect of glucose). In contrast, the effect of PA on ISR was almost 3 times that by glucose, suggesting that the metabolism of PA is not the dominant mechanism mediating PA\'s effect on ISR. This was further supported by lack of inhibition of PA-stimulated OCR and ISR when blocking entry of PA into (with etomoxir), and PA\'s lack of stimulation of reduced cytochrome c in the presence of high glucose. Consistent with the lack of metabolic stimulation by PA, an inhibitor of calcium release from the endoplasmic reticulum, but not a blocker of L-type calcium channels, abolished the PA-induced elevation of cytosolic calcium. Notably, ISR was unaffected by thapsigargin showing the dissociation of endoplasmic reticulum calcium release and second phase insulin secretion. In conclusion, stimulation of ISR by PA was mediated by mechanisms largely independent of the oxidation of the fuel.

Keyword: mitochondria

[The insulin initiates "kinetic perfection" of biologic function of locomotion. The glucose as substratum for synthesis of ω-9 oleic fatty by cross-striated miocytes.]

The phylogenesis theory affords ground for the following propositions. 1. There is no absorption of glucose from intercellular medium by cells in vivo until there is possibility to absorb polar fatty acids from associates with albumin. 2. The late in phylogenesis humoral insulin regulates no stages of glucose metabolism; they are completed a billion years before hormone synthesis. 3. The phylogenetically late insulin is "hostage" of biological function of trophology, function of nutrition, biological reaction of exotrophy; it has no possibility to decrease in food excessed amount of physiologic saturated fatty acids with low kinetic parameters of β-oxidation in . 4. The early in phylogenesis, resistant organizational to insulin pool of visceral fatty cells of omentum and late pool of insulin-dependent adipocytes are different in many functional parameters. 5. All "metabolic pandemics" such as syndrome of resistance to insulin, atherosclerosis, metabolic arterial hypertension, metabolic syndrome and obesity are primarily pathologies of fatty acids. 6. All "metabolic pandemics" are pathologies of one biological function, function of locomotion under single algorithm of formation of their pathogenesis. 7. The etiological factor of "metabolic pandemics" is uniform - effect of environmental factors in form of disorder of biological function of trophology, function of nutrition; aphysiological excess content in food of saturated fatty , aphysiological trans-forms of fatty acids and ω-7-palmitoleic mono unsaturated fatty . The insulin activates absorption by myocytes, cardiomyocytes of glucose as substrate of synthesis out of it in situ de novo ω-9 oleic mono unsaturated fatty . With such physical chemical parameters that oxidize it with the most high constant of velocity of reaction and high effectiveness of formation of ATP.

Keyword: mitochondria

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to , and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: mitochondria

Nicotinamide riboside regulates inflammation and mitochondrial markers in AML12 hepatocytes.

The NAD precursor nicotinamide riboside (NR) is a type of vitamin B found in cow\'s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer\'s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes.A subset of hepatocytes was treated with (PA; 250 µM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 µM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed.Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes.These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.

Keyword: mitochondria

-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.

Can we use a mitochondrial-targeted antioxidant (Mitoquinone) during in vitro embryo culture to rescue developmental competence of oocytes matured under lipotoxic conditions, exhibiting mitochondrial dysfunction and oxidative stress?Supplementation of embryo culture media with Mitoquinone reduced oxidative stress and prevented mitochondrial uncoupling in embryos derived from metabolically compromised oocytes in vitro, leading to higher blastocyst rates and lower blastomeric apoptosis.Maternal metabolic disorders, such as obesity and type-II diabetes are associated with hyperlipidemia and elevated free fatty (FFA) concentrations in the ovarian follicular fluid (FF). Oocyte maturation under these lipotoxic conditions results in increased oxidative stress levels, mitochondrial dysfunction, reduced developmental competence and disappointing IVF results.A well-described bovine oocyte IVM model was used, where a pathophysiologically relevant elevated FF concentrations of (PA; 150\xa0μM or 300\xa0μM) were added to induce oxidative stress. After fertilization (Day 0, D0), zygotes were in vitro cultured (IVC, from D1 to D8) in standard fatty -free media in the presence or absence of Mitoquinone or its carrier triphenyl-phosphonium.Embryo cleavage and fragmentation (D2) and blastocyst rates (D8) were recorded. Mitochondrial activity and oxidative stress in cleaved embryos at D2 were determined using specific fluorogenic probes and confocal microscopy. D8 blastocysts were used to (i) examine the expression of marker genes related to mitochondrial unfolded protein responses (UPRmt; HSPD1 and HSPE1), mitochondrial biogenesis (TFAM), endoplasmic reticulum (ER) UPR (ATF4, ATF6 and BiP) and oxidative stress (CAT, GPX1 and SOD2) using real time RT-PCR; (ii) determine cell differentiation and apoptosis using CDX-2 and cleaved caspase-3 immunostaining; and (iii) measure mtDNA copy numbers. This was tested in a series of experiments with at least three independent replicates for each, using a total of 2525 oocytes. Differences were considered significant if a P value was <0.05 after Bonferroni correction.Exposure to PA during IVM followed by culture under control conditions resulted in a significant increase in oxidative stress in embryos at D2. This was associated with a significant reduction in mitochondrial inner membrane potential (uncoupling) compared with solvent control (P\xa0<\u20090.05). The magnitude of these effects was PA-concentration dependent. Consequently, development to the blastocysts stage was significantly hampered. Surviving blastocysts exhibited high apoptotic cell indices and upregulated mRNA expression indicating persistent oxidative stress, mitochondrial and ER UPRs. In contrast, supplementation of PA-derived zygotes with Mitoquinone during IVC (i) prevented mitochondrial uncoupling and alleviated oxidative stress at D2; and (ii) rescued blastocyst quality; normalized oxidative stress and UPR related genes and apoptotic cell indices (P\xa0>\u20090.01 compared with solvent control). Mitoquinone also improved blastocyst rate in PA-exposed groups, an effect that was dependent on PA concentration.N/A.This is a fundamental study performed using a bovine in vitro model using PA-induced lipotoxicity during oocyte maturation. PA is the most predominant FFA in the FF that is known to induce lipotoxicity; however, in vivo maturation in patients suffering from maternal metabolic disorders involve more factors that cannot be represented in one model. Nevertheless, focusing on the carryover oxidative stress as a known key factor affecting developmental competence, and considering the novel beneficial rescuing effects of Mitoquinone shown here, we believe this model is of high biological relevance.Human oocytes collected for IVF treatments from patients with maternal metabolic disorders are vulnerable to lipotoxicity and oxidative stress during in vivo maturation. The results shown here suggest that mitochondrial targeted therapy, such as using Mitoquinone, during IVC may rescue the developmental competence and quality of these compromised oocytes. After further clinical trials, this may be a valuable approach to increase IVF success rates for infertile patients experiencing metabolic disorders.This study was financially supported by a BOF/KP grant number 34399, from the University of Antwerp, Belgium. W.F.A.M. was supported by a postdoctoral fellowship from the Research Foundation-Flanders (FWO), grant number 12I1417N, Antwerp, Belgium. The Leica SP 8 confocal microscope used in this study was funded by the Hercules Foundation of the Flemish Government (Hercules grant AUHA.15.12). All authors have no financial or non-financial competing interests to declare.© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: mitochondria

[The influence of spermine on Ca(2+)-dependent permeability transition in and liposomes induced by and α,Ω-hexadecanedioic acids].

The effect of spermine on Ca(2+)-dependent permeability transition in and liposomes induced by and α,Ω-hexadecanedioic was studied. It has been shown that spermine inhibited the cyclosporin A-insensitive mitochondrial swelling induced by and Ca2+ and α,Ω-hexadecanedioic and Ca2+. 100 μM spermine did not influence the mitochondrial respiration in state V2 and the respiration stimulated by , α,Ω-hexadecanedioic and Ca2+. Pre-incubation of liposomes with 100 μM spermine resulted in the inhibition of /Ca(2+)- and α,Ω-hexadecanedioic /Ca(2+)-induced release of the fluorescent dye sulforhodamine B from liposomes. At the same time, spermine added to fatty acids-contained membranes of liposomes stimulated Ca(2+)-dependent release of sulforhodamine B from liposomes. It was shown that an addition of spermine to liposomes resulted in a significant increase in z-potential of liposomal membranes (from -39.8 mV to -18.6 mV). A possible mechanism of spermine influence on /Ca(2+)- and α,Ω-hexadecanedioic /Ca(2+)-induced permeability transition in and liposomes is discussed.

Keyword: mitochondria

Dynamic Regulation of Long-Chain Fatty Oxidation by a Noncanonical Interaction between the MCL-1 BH3 Helix and VLCAD.

MCL-1 is a BCL-2 family protein implicated in the development and chemoresistance of human cancer. Unlike its anti-apoptotic homologs, Mcl-1 deletion has profound physiologic consequences, indicative of a broader role in homeostasis. We report that the BCL-2 homology 3 (BH3) α helix of MCL-1 can directly engage very long-chain acyl-CoA dehydrogenase (VLCAD), a key enzyme of the mitochondrial fatty β-oxidation (FAO) pathway. Proteomic analysis confirmed that the mitochondrial matrix isoform of MCL-1 (MCL-1) interacts with VLCAD. Mcl-1 deletion, or eliminating MCL-1 alone, selectively deregulated long-chain FAO, causing increased flux through the pathway in response to\xa0nutrient deprivation. Transient elevation in MCL-1 upon serum withdrawal, a striking increase in MCL-1 BH3/VLCAD interaction upon titration, and direct modulation of enzymatic activity by the MCL-1 BH3 α helix are consistent with dynamic regulation. Thus, the MCL-1 BH3 interaction with VLCAD revealed a separable, gain-of-function role for MCL-1 in the regulation of lipid metabolism.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Transport of Ca and Ca-Dependent Permeability Transition in Rat Liver under the Streptozotocin-Induced Type I Diabetes.

Although diabetes mellitus is known to be a disease associated with mitochondrial dysfunction, not everything is clear about mitochondrial Ca transport and Ca-induced permeability transition in diabetic cells. The objective of this work was to study the operation of MCU and Ca-dependent mitochondrial permeabilization in the liver cells of Sprague-Dawley rats under the streptozotocin-induced type I diabetes. It was shown that two weeks after the induction of diabetes, the rate of Ca uptake by the of diabetic animals increased ~1.4-fold. The expression of MCU and MICU1 subunits did not change, yet the quantity of dominant-negative MCUb channel subunits was almost twice as lower. The organelles also became more resistant to the induction of CsA-sensitive MPT pore and less resistant to the induction of CsA-insensitive palmitate/Ca-induced pore. The of diabetic liver cells also showed changes in the lipid matrix of their membranes. The content of fatty acids in the membranes grew, and microviscosity of the lipid bilayer (assessed with laurdan) increased. At the same time, lipid peroxidation (assessed by the production of malonic dialdehyde) was stimulated. The paper discusses the consequences of the diabetes-related changes in in the context of cell physiology.

Keyword: mitochondria

The mother of all endocytosis.

Massive endocytosis is initiated by a series of steps that involve a sudden influx of calcium ions, changes in , and modification of surface proteins by lipids. A better understanding of this process could lead to new approaches to reducing the tissue damage that is caused by heart attacks.

Keyword: mitochondria

Palmitate-induced impairment of glucose-stimulated insulin secretion precedes mitochondrial dysfunction in mouse pancreatic islets.

It has been well established that excessive levels of glucose and palmitate lower glucose-stimulated insulin secretion (GSIS) by pancreatic β-cells. This β-cell \'glucolipotoxicity\' is possibly mediated by mitochondrial dysfunction, but involvement of bioenergetic failure in the pathological mechanism is the subject of ongoing debate. We show in the present study that increased palmitate levels impair GSIS before altering mitochondrial function. We demonstrate that GSIS defects arise from increased insulin release under basal conditions in addition to decreased insulin secretion under glucose-stimulatory conditions. Real-time respiratory analysis of intact mouse pancreatic islets reveals that mitochondrial ATP synthesis is not involved in the mechanism by which basal insulin is elevated. Equally, mitochondrial lipid oxidation and production of reactive oxygen species (ROS) do not contribute to increased basal insulin secretion. Palmitate does not affect KCl-induced insulin release at a basal or stimulatory glucose level, but elevated basal insulin release is attenuated by palmitoleate and associates with increased intracellular calcium. These findings deepen our understanding of β-cell glucolipotoxicity and reveal that palmitate-induced GSIS impairment is disconnected from mitochondrial dysfunction, a notion that is important when targeting β-cells for the treatment of diabetes and when assessing islet function in human transplants.© 2016 Authors; published by Portland Press Limited.

Keyword: mitochondria

HADHA, the alpha subunit of the mitochondrial trifunctional protein, is involved in long-chain fatty -induced autophagy in intestinal epithelial cells.

Genome-wide association studies have identified autophagy-related susceptibility genes for inflammatory bowel disease (IBD); however, whether autophagy regulators can be utilized as therapeutic targets remains unclear. To identify novel microtubule-associated protein 1 light chain 3 (LC3)-interacting proteins in intestinal epithelial cells (IECs), we isolated primary IECs from green fluorescent protein (GFP)-LC3 mice. We performed immunoprecipitation with a GFP antibody and then analyzed co-immunoprecipitates by mass spectrometry. HADHA was identified as an LC3-interacting protein from primary IECs. The HADHA gene encodes the alpha subunit of the mitochondrial trifunctional protein. Given that HADHA catalyzes the last three steps of mitochondrial beta-oxidation of long-chain fatty acids, we investigated whether long-chain fatty acids induce autophagy in IECs. We found that induced autophagy in DLD-1, HT29, and HCT116\xa0cells. HADHA was expressed in not only the but also the cytosol. LC3 puncta co-localized with HADHA, which were enhanced by stimulation. However, LC3 puncta did not co-localize with Tom20, suggesting that HADHA was induced to associate with LC3 puncta at sites other than the . Thus, HADHA may have extra-mitochondrial functions. Furthermore, we found that induced cell death in IECs, which was accelerated by bafilomycin A and chloroquine. These findings suggested that -induced autophagy supports the survival of IECs. Taken together, these results suggested that HADHA is involved in long-chain fatty -induced autophagy in IECs, thus providing new insights into the pathology of IBD and revealing novel therapeutic targets of IBD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

stimulates interleukin-8 via the TLR4/NF-κB/ROS pathway and induces mitochondrial dysfunction in bovine oviduct epithelial cells.

We investigated the effect of (PA), a major saturated fatty in NEFA, on bovine oviduct epithelial cells (OECs) during in vitro cell culture.Bovine oviductal tissues ipsilateral to the corpus luteum were collected 1-3\xa0days after ovulation; the OECs were isolated and cultured.PA increased lipid accumulation and activated caspase-3 in OECs, resulting in decreased cell proliferation. PA also stimulated the secretion of inflammatory cytokine interleukin (IL)-8 depending on TLR4, NF-κB activation, and reactive oxygen species (ROS) production. Moreover, PA induced mitochondrial dysfunction, including mitochondrial fission, ATP production, and mitochondrial ROS production. It also increased levels of LC3 and p62 proteins, suggesting autophagy induction in OECs.We suggest that bovine OECs recognize an excessive increase in endogenous and sterile danger signals, such as PA, which may contribute to chronic oviductal inflammation, resulting in infertility associated with oviductal dysfunction.© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: mitochondria

Synergistic effects of zinc oxide nanoparticles and Fatty acids on toxicity to caco-2 cells.

Fatty acids exposure may increase sensitivity of intestinal epithelial cells to cytotoxic effects of zinc oxide (ZnO) nanoparticles (NPs). This study evaluated the synergistic effects of ZnO NPs and (PA) or free fatty acids (FFAs) mixture (oleic/PA 2:1) on toxicity to human colon epithelial (Caco-2) cells. The ZnO NPs exposure concentration dependently induced cytotoxicity to Caco-2 cells showing as reduced proliferation and activity measured by 3 different assays. PA exposure induced cytotoxicity, and coexposure to ZnO NPs and PA showed the largest cytotoxic effects. The presence of FFAs mixture did not affect the ZnO NPs-induced cytotoxicity. Filtration of freshly prepared suspension of NPs through a 0.45-µm pore size membrane significantly reduced the cytotoxicity, indicating a role of concentration or size of particles in cytotoxic effects. The ZnO NPs and PA coexposure induced production of mitochondrial reactive oxygen species (mROS) but not intracellular ROS production, whereas FFAs mixture exposure did not induce mROS and inhibited intracellular ROS. Both ZnO NPs and fatty acids (PA and FFAs mixture) promoted lysosomal destabilization, which was not correlated with cytotoxicity. These results indicated that PA can enhance ZnO NPs-induced cytotoxicity probably by the augmentation of mROS production, whereas FFAs mixture did not affect ROS production. Synergistic effects between ZnO NPs and fatty acids may be important when considering NPs toxicity via oral exposure.© The Author(s) 2014.

Keyword: mitochondria

Fasting rapidly increases fatty oxidation in white adipose tissue of young broiler chickens.

Upregulating the fatty oxidation capacity of white adipose tissue in mice protects against diet-induced obesity, inflammation and insulin resistance. Part of this capacity results from induction of brown-like adipocytes within classical white depots, making it difficult to determine the oxidative contribution of the more abundant white adipocytes. Avian genomes lack a gene for uncoupling protein 1 and are devoid of brown adipose cells, making them a useful model in which to study white adipocyte metabolism in vivo. We recently reported that a brief (5\xa0hour) period of fasting significantly upregulated many genes involved in mitochondrial and peroxisomal fatty oxidation pathways in white adipose tissue of young broiler chickens. The objective of this study was to determine if the effects on gene expression manifested in increased rates of fatty oxidation. Abdominal adipose tissue was collected from 21\xa0day-old broiler chicks that were fasted for 3, 5 or 7\xa0hours or fed ad libitum (controls). Fatty oxidation was determined by measuring and summing CO production and C-labeled -soluble metabolites from the oxidation of [1-C] . Fasting induced a progressive increase in complete fatty oxidation and citrate synthase activity relative to controls. These results confirm that fatty oxidation in white adipose tissue is dynamically controlled by nutritional status. Identifying the underlying mechanism may provide new therapeutic targets through which to increase fatty oxidation in situ and protect against the detrimental effects of excess free fatty acids on adipocyte insulin sensitivity.

Keyword: mitochondria

Fatty activates NLRP3 inflammasomes in mouse Kupffer cells through mitochondrial DNA release.

Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease in many developed and developing countries worldwide. It has been well established that the chronic sterile inflammation caused by the NLRP3 inflammasome is closely related to NAFLD development. Kupffer cells (KCs) are involved in the pathogenesis of various liver diseases. We used methionine choline-deficient diets to establish a mouse nonalcoholic steatohepatitis (NASH) model. The expression and formation of the NLRP3 inflammasome in the KCs from the mouse and cell models were determined by Western blotting and co-immunoprecipitation. Evidence of mitochondrial DNA (mtDNA) release was determined by live cell labeling and imaging. KCs and the NLRP3 inflammasome exerted proinflammatory effects on the development and progression of NASH through secretion of the proinflammatory cytokine IL-1β. NLRP3, ASC and Caspase-1 protein expression levels in KCs from NASH mouse livers were significantly higher than those in KCs from NLRP3 mice, and the number of NLRP3 inflammasome protein complexes was significantly higher in KCs from NASH mouse livers, whereas these protein complexes could not be formed in NLRP3 mice. In in vitro experiments, (PA) decreased the mitochondrial membrane potential and subsequently induced mtDNA release from the to the cytoplasm. NLRP3 inflammasome expression was substantially increased, and mtDNA-NLRP3 inflammasome complexes formed upon PA stimulation. Our data suggest that mtDNA released from during PA stimulation causes NLRP3 inflammasome activation, providing a missing link between NLRP3 inflammasome activation and NASH development, via binding of cytosolic mtDNA to the NLRP3 inflammasome.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Inhibition of autophagic turnover in β-cells by fatty acids and glucose leads to apoptotic cell death.

Autophagy, a cellular recycling process responsible for turnover of cytoplasmic contents, is critical for maintenance of health. Defects in this process have been linked to diabetes. Diabetes-associated glucotoxicity/lipotoxicity contribute to impaired β-cell function and have been implicated as contributing factors to this disease. We tested the hypothesis that these two conditions affect β-cell function by modulating autophagy. We report that exposure of β-cell lines and human pancreatic islets to high levels of glucose and lipids blocks autophagic flux and leads to apoptotic cell death. EM analysis showed accumulation of autophagy intermediates (autophagosomes), with abundant engulfed cargo in (PA)- or glucose-treated cells, indicating suppressed autophagic turnover. EM studies also showed accumulation of damaged , endoplasmic reticulum distention, and vacuolar changes in PA-treated cells. Pulse-chase experiments indicated decreased protein turnover in β-cells treated with PA/glucose. Expression of mTORC1, an inhibitor of autophagy, was elevated in β-cells treated with PA/glucose. mTORC1 inhibition, by treatment with rapamycin, reversed changes in autophagic flux, and cell death induced by glucose/PA. Our results indicate that nutrient toxicity-induced cell death occurs via impaired autophagy and is mediated by activation of mTORC1 in β-cells, contributing to β-cell failure in the presence of metabolic stress.© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: mitochondria

Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis.

Mitochondrial dysfunction plays a crucial role in the development of non-alcoholic steatohepatitis (NASH). However, the regulator of mitochondrial dysfunction in the pathogenesis of NASH is still largely unclear. CXCR3 is an essential pro-inflammatory factor in chronic liver diseases. We explored the significance of CXCR3 in regulating mitochondrial function during NASH development in animal models and cultured hepatocytes.The effects of CXCR3 on mitochondrial function were evaluated by genetic knockout or pharmacological inhibition in mouse models and . The ultrastructural changes of were assessed by transmission electron microscopy (TEM). Hepatic levels of mitochondrial reactive oxygen species (ROS), DNA damage, membrane potential and ATP were examined.CXCR3 ablation by genetic knockout or pharmacological inhibition in mice protected against NASH development by influencing mitochondrial function. Similarly, depletion of CXCR3 reduced steatohepatitis injury in cultured hepatocytes. TEM analysis revealed that liver mitochondrial integrity was much improved in CXCR3 knockout (CXCR3) compared to wildtype (WT) mice. In agreement with this, impaired mitochondrial function was pronounced in WT mice compared to CXCR3 mice, evidenced by increased protein expression of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and decreased protein expression of mitofusin-1 (MFN1). Mitochondrial dysfunction was induced in AML-12 hepatocytes by methionine and choline deficient medium and in HepG2 cells by . The impaired mitochondrial function in both cell lines was evidenced by reduced membrane potential and ATP content, and by increased mitochondrial ROS accumulation and DNA damage. However, CXCR3 knockdown by siCXCR3 significantly diminished the mitochondrial dysfunction in both AML-12 and HepG2 hepatocytes. In addition, inhibition of CXCR3 by CXCR3 specific antagonists SCH546738 and AMG487 restored mitochondrial function and inhibited mitochondrial-dependent apoptosis in the liver of WT mice fed with methionine and choline deficient diet.CXCR3 induces mitochondrial dysfunction, which contributes to the pathogenesis of steatohepatitis. Pharmacologic blockade of CXCR3 prevents mitochondrial dysfunction and restores the severity of steatohepatitis, indicating a potential clinical impact for controlling the disease.

Keyword: mitochondria

Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic fatty liver disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high-fat diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of fatty liver disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Alpha-linolenic protects the developmental capacity of bovine cumulus-oocyte complexes matured under lipotoxic conditions in vitro.

Elevated concentrations of free fatty acids (FFAs), predominantly , stearic, and oleic acids (PSO), exert detrimental effects on oocyte developmental competence. This study examined the effects of omega-3 alpha-linolenic (ALA) during in vitro oocyte maturation (IVM) in the presence of PSO on subsequent embryo development and quality, and the cellular mechanisms that might be involved. Bovine cumulus-oocyte complexes (COCs) were supplemented during IVM with ALA (50 μM), PSO (425 μM), or PSO+ALA. Compared with FFA-free controls (P\xa0<\xa00.05), PSO increased embryo fragmentation and decreased good quality embryos on day 2 postfertilization. Day 7 blastocyst rate was also reduced. Day 8 blastocysts had lower cell counts and higher apoptosis but normal metabolic profile. In the PSO group, cumulus cell (CC) expansion was inhibited with an increased CC apoptosis while COC metabolism was not affected. Mitochondrial inner membrane potential (MMP; JC-1 staining) was reduced in the CCs and oocytes. Heat shock protein 70 (HSP70) but not glucose-regulated protein 78 kDa (GRP78, known as BiP; an endoplasmic reticulum stress marker) was upregulated in the CCs. Higher reactive oxygen species levels (DCHFDA staining) were detected in the oocytes. In contrast, adding ALA in the presence of PSO normalized embryo fragmentation, cleavage, blastocyst rates, and blastocyst quality compared to controls (P\xa0>\xa00.05). Combined treatment with ALA also reduced CC apoptosis, partially recovered CC expansion, abrogated the reduction in MMP in the CCs but not in the oocytes, and reduced BiP and HSP70 expression in CCs, compared with PSO only (P\xa0<\xa00.05). In conclusion, ALA supplementation protected oocyte developmental capacity under lipotoxic conditions mainly by protecting cumulus cell viability.© The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: mitochondria

Hepatocellular toxicity of benzbromarone: effects on mitochondrial function and structure.

Benzbromarone is an uricosuric structurally related to amiodarone and a known mitochondrial toxicant. The aim of the current study was to improve our understanding in the molecular mechanisms of benzbromarone-associated hepatic mitochondrial toxicity. In HepG2 cells and primary human hepatocytes, ATP levels started to decrease in the presence of 25-50μM benzbromarone for 24-48h, whereas cytotoxicity was observed only at 100μM. In HepG2 cells, benzbromarone decreased the mitochondrial membrane potential starting at 50μM following incubation for 24h. Additionally, in HepG2 cells, 50μM benzbromarone for 24h induced mitochondrial uncoupling,and decreased mitochondrial ATP turnover and maximal respiration. This was accompanied by an increased lactate concentration in the cell culture supernatant, reflecting increased glycolysis as a compensatory mechanism to maintain cellular ATP. Investigation of the electron transport chain revealed a decreased activity of all relevant enzyme complexes. Furthermore, treatment with benzbromarone was associated with increased cellular ROS production, which could be located specifically to . In HepG2 cells and in isolated mouse liver , benzbromarone also reduced metabolism due to an inhibition of the long-chain acyl CoA synthetase. In HepG2 cells, benzbromarone disrupted the mitochondrial network, leading to mitochondrial fragmentation and a decreased mitochondrial volume per cell. Cell death occurred by both apoptosis and necrosis. The study demonstrates that benzbromarone not only affects the function of in HepG2 cells and human hepatocytes, but is also associated with profound changes in mitochondrial structure which may be associated with apoptosis.Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Keyword: mitochondria

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high- diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut microbiota and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: mitochondria

[The nicotinic as an inhibitor of lipolysis in the phylogenetically early visceral cells; insulin also blocks hydrolysis of triglycerides in the phylogenetically late adipocytes.]

It is valid to consider effect of nicotinic as an insulin-mimetic one. The uniformity of biologic effect of exogenous nicotinic and endogenous insulin permits to become aware that a) the hypo-lipidemic activity of insulin, inhibition of lipolysis in phylogenetically late insulin-dependent adipocytes and decreasing of content of unesterified fatty acids in blood plasma are considered as a basis of hypoglycemic effect of insulin; b) nicotinic similar to insulin blocks lipolysis too but in hormoneindependent visceral fatty cells. The counter-insular effect is manifested in vivo by exogenous and endogenous saturated fatty by force of physical chemical characteristics. The biological role of insulin consists in regulation of metabolism of fatty acids mainly unesterified fatty acids and in absorption of glucose by all insulin-dependent cells. It is supposed that contractile cells (myocytes and cardiomyocytes) cumulate glycogen for implementing glucose as a substrate in synthesis in situ de novo of ῲ-9 oleic mono-saturated fatty . The insulin initiates synthesis because process this mono-saturated fatty to β-oxidation with the highest constant of reaction velocity. This is conditioned by physical chemical characteristics of oleic mono-saturated fatty , positioning of double bond in the chain of fatty ; this ensures maximal efficiency of ATP gaining. In phylogenesis, the low efficient alternative of metabolism of fatty acids is formed the earliest. At the later stages of phylogenesis the cells, under becoming of motion function, worked out more efficient alternative of ATP synthesis from exogenous ῲ-9 oleic monosaturated fatty . At the late stages of phylogenesis, under becoming of biological function of locomotion (motion at the expense of contraction of cross-striated skeletal myocytes) insulin began to activate absorption of glucose by cells with subsequent synthesis of endogenous ῲ-9 oleic mono-saturated fatty out of it. The aphysiological effect of environment factors in the form of derangement of biological function of trophology (feeding), surplus of unesterified fatty acids in food serve as a main cause of such a high rate of functional derangement - insulin resistance syndrome in populations. Hence, there is no reason to call it diabetes mellitus type II.

Keyword: mitochondria

Adaptations of hepatic lipid metabolism and in dairy cows with mild fatty liver.

The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. Fatty liver is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and in dairy cows with mild fatty liver, liver and blood samples were collected from healthy cows (n = 15) and cows with mild fatty liver (n = 15). To determine the effects of acids (PA), one of the major component of fatty acids, on lipid metabolism and in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM). Dairy cows with mild fatty liver displayed hepatic lipid accumulation. The protein levels of sterol regulatory element-binding protein 1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPARα) and mRNA levels of acetyl CoA carboxylase 1 (ACC1), fatty synthase (FAS), acyl-CoA oxidase (ACO), and carnitine palmitoyltransferase 1A (CPT1A) were significantly higher in dairy cows with mild fatty liver than in control cows. The hepatic mitochondrial DNA content, mRNA levels of oxidative phosphorylation complexes I to V (CO 1-V), protein levels of cytochrome c oxidase subunit IV (COX IV), voltage dependent anion channel 1 (VDAC1), peroxisome proliferator activated receptor-γ coactivator-1α (PGC-1α) and nuclear respiratory factor 1 (NRF1), and adenosine triphosphate (ATP) content were all markedly increased in the liver of dairy cows with mild fatty liver compared with healthy cows. The PA treatment significantly increased lipid accumulation; protein levels of SREBP-1c and PPARα; and mRNA levels of ACC1, FAS, ACO, and CPT1A in calf hepatocytes. Moreover, the mitochondrial DNA content, mRNA levels of CO 1-V, protein levels of COX IV, VDAC1, PGC-1α, NRF1, mitochondrial transcription factor A, and ATP content were significantly increased in PA-treated hepatocytes compared with control hepatocytes. The protein level of mitofusin-2 was significantly decreased in PA-treated groups. In conclusion, lipid synthesis and oxidation, number of , and ATP production were increased in the liver of dairy cows with mild fatty liver and PA-treated calf hepatocytes. These changes in hepatic and lipid metabolism may be the adaptive mechanism of dairy cows with mild fatty liver.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: mitochondria

stimulates energy metabolism and inhibits insulin/PI3K/AKT signaling in differentiated human neuroblastoma cells: The role of mTOR activation and mitochondrial ROS production.

The high consumption of saturated lipids has been largely associated with the increasing prevalence of metabolic diseases. In particular, saturated fatty acids such as (PA) have been implicated in the development of insulin resistance in peripheral tissues. However, how neurons develop insulin resistance in response to lipid overload is not fully understood. Here, we used cultured rat cortical neurons and differentiated human neuroblastoma cells to demonstrate that PA blocks insulin-induced metabolic activation, inhibits the activation of the insulin/PI3K/Akt pathway and activates mTOR kinase downstream of Akt. Despite the fact that fatty acids are not normally used as a significant source of fuel by neural cells, we also found that short-term neuronal exposure to PA reduces the NAD/NADH ratio, indicating that PA modifies the neuronal energy balance. Finally, inhibiting mitochondrial ROS production with mitoTEMPO prevented the deleterious effect of PA on insulin signaling. This work provides novel evidence of the mechanisms behind saturated fatty -induced insulin resistance and its metabolic consequences on neuronal cells.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

dysregulates the Hippo-YAP pathway and inhibits angiogenesis by inducing mitochondrial damage and activating the cytosolic DNA sensor cGAS-STING-IRF3 signaling mechanism.

Impaired angiogenesis and wound healing carry significant morbidity and mortality in diabetic patients. Metabolic stress from hyperglycemia and elevated free fatty acids have been shown to inhibit endothelial angiogenesis. However, the underlying mechanisms remain poorly understood. In this study, we show that dysregulation of the Hippo-Yes-associated protein (YAP) pathway, an important signaling mechanism in regulating tissue repair and regeneration, underlies (PA)-induced inhibition of endothelial angiogenesis. PA inhibited endothelial cell proliferation, migration, and tube formation, which were associated with increased expression of mammalian Ste20-like kinases 1 (MST1), YAP phosphorylation/inactivation, and nuclear exclusion. Overexpression of YAP or knockdown of MST1 prevented PA-induced inhibition of angiogenesis. When searching upstream signaling mechanisms, we found that PA dysregulated the Hippo-YAP pathway by inducing mitochondrial damage. PA treatment induced mitochondrial DNA (mtDNA) release to cytosol, and activated cytosolic DNA sensor cGAS-STING-IRF3 signaling. Activated IRF3 bound to the gene promoter and induced MST1 expression, leading to MST1 up-regulation, YAP inactivation, and angiogenesis inhibition. Thus, mitochondrial damage and cytosolic DNA sensor cGAS-STING-IRF3 signaling are critically involved in PA-induced Hippo-YAP dysregulation and angiogenesis suppression. This mechanism may have implication in impairment of angiogenesis and wound healing in diabetes.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: mitochondria

Modest decrease in PGC1α results in TAG accumulation but not in insulin resistance in L6 myotubes.

PGC-1α is an important cellular protein (coactivator) regulating myocyte number and function, and therefore whole cellular energy status. The aim of this work was to investigate the effects of modest, temporary PGC-1α knock-down on L6 myotubes insulin resistance in a light of cellular lipid metabolism.Gas liquid chromatography was applied for assessing FAs content and composition. For the expression of mitochondrial enzymes, as well as FA and glucose transporters, Western Blot technique was adopted. Additionally, radiolabelled glucose and uptake was performed to estimate the nutrients cellular influx.Modest (-24%) PGC-1α protein ablation resulted in decreased mitochondrial activity in general (reduced Cyt C content) and FAs oxidation in particular (diminished β-HAD expression) without increased FAs cellular influx. The aforementioned intervention led to significantly increased TAG cellular level, but not DAG nor CER. Consequently, no changes in cellular insulin responsiveness were noticed.Modest (-24%) PGC-1α protein depletion results in lipid accumulation, without causing insulin resistance. Importantly, it seems that this TAG loading is a result of decreased mitochondrial oxidative capacity and/or possibly increased lipid biosynthesis but not fatty cellular influx.© 2015 S. Karger AG, Basel.

Keyword: mitochondria

Calcium Uptake via Mitochondrial Uniporter Contributes to -Induced Apoptosis in Mouse Podocytes.

Podocytes are component cells of the glomerular filtration barrier, and their loss by apoptosis is the main cause of proteinuria that leads to diabetic nephropathy (DN). Therefore, insights into podocyte apoptosis mechanism would allow a better understanding of DN pathogenesis and thus help develop adequate therapeutic strategies. Here, we investigated the molecular mechanism of -inhibited cell death in mouse podocytes, and found that increased cell death in a dose- and time-dependent manner. induces apoptosis in podocytes through upregulation of cytosolic and mitochondrial Ca , mitochondrial membrane potential (MMP), cytochrome c release, and depletion of endoplasmic reticulum (ER) Ca . The intracellular calcium chelator, 1,2-bis (2-aminophenoxy) ethane-N,N,N, N\'-tetraacetic tetrakis acetoxymethyl ester (BAPTA-AM), partially prevented this upregulation whereas 2-aminoethoxydiphenyl borate (2-APB), an inositol 1,4,5-triphosphate receptor (IP3R) inhibitor; dantrolene, a ryanodine receptor (RyR) inhibitor; and 4,4\'-diisothiocyanatostibene-2,2\'-disulfonic (DIDS), an anion exchange inhibitor, had no effect. Interestingly, ruthenium red and Ru360, both inhibitors of the mitochondrial Ca uniporter (MCU), blocked -induced mitochondrial Ca elevation, cytochrome c release from to cytosol, and apoptosis. siRNA to MCU markedly reduced -induced apoptosis. These data indicate that Ca uptake via mitochondrial uniporter contributes to -induced apoptosis in mouse podocytes. J. Cell. Biochem. 118: 2809-2818, 2017.© 2017 Wiley Periodicals, Inc.

Keyword: mitochondria

Biochemical precursor effects on the fatty production in cell suspension cultures of Theobroma cacao L.

Cocoa butter (CB) is composed of 96% , stearic, oleic, linoleic and linolenic fatty acids that are responsible for the hardness, texture and fusion properties of chocolate. Through in\xa0vitro plant cell culture it is possible to modify CB lipid profiles and to study the fatty biosynthesis pathway on a subcellular level, evaluating fundamental aspects to enhance in\xa0vitro fatty production in a specific and controlled way. In this research, culture media was supplemented with acetate, biotin, pyruvate, bicarbonate and glycerol at three different concentrations and the effects on the biomass production (g/L), cell viability, and fatty acids profile and production was evaluated in in\xa0vitro cell suspensions culture. It was found that biotin stimulated fatty synthesis without altering cell viability and cell growth. It was also evident a change in the lipid profile of cell suspensions, increasing middle and long chain fatty acids proportion, which are unusual to those reported in seeds; thus implying that it is possible to modify lipid profiles according to the treatment used. According to the results of sucrose gradients and enzyme assays performed, it is proposed that cacao cells probably use the pentose phosphate pathway, being the key organelle in the carbon flux for the synthesis of reductant power and fatty precursors.Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Keyword: mitochondria

Overexpression of PGC-1α increases peroxisomal activity and mitochondrial fatty oxidation in human primary myotubes.

Peroxisomes are indispensable organelles for lipid metabolism in humans, and their biogenesis has been assumed to be under regulation by peroxisome proliferator-activated receptors (PPARs). However, recent studies in hepatocytes suggest that the mitochondrial proliferator PGC-1α (peroxisome proliferator-activated receptor gamma coactivator-1α) also acts as an upstream transcriptional regulator for enhancing peroxisomal abundance and associated activity. It is unknown whether the regulatory mechanism(s) for enhancing peroxisomal function is through the same node as mitochondrial biogenesis in human skeletal muscle (HSkM) and whether fatty oxidation (FAO) is affected. Primary myotubes from vastus lateralis biopsies from lean donors (BMI = 24.0 ± 0.6 kg/m; = 6) were exposed to adenovirus encoding human PGC-1α or GFP control. Peroxisomal biogenesis proteins (peroxins) and genes () responsible for proliferation and functions were assessed by Western blotting and real-time qRT-PCR, respectively. [1-C] and [1-C]lignoceric (exclusive peroxisomal-specific substrate) were used to assess mitochondrial oxidation of peroxisomal-derived metabolites. After overexpression of PGC-1α, ) peroxisomal membrane protein 70 kDa (PMP70), PEX19, and mitochondrial citrate synthetase protein content were significantly elevated ( < 0.05), ) , , key , and peroxisomal β-oxidation mRNA expression levels were significantly upregulated ( < 0.05), and ) a concomitant increase in lignoceric oxidation by both peroxisomal and mitochondrial activity was observed ( < 0.05). These novel findings demonstrate that, in addition to the proliferative effect on , PGC-1α can induce peroxisomal activity and accompanying elevations in long-chain and very-long-chain fatty oxidation by a peroxisomal-mitochondrial functional cooperation, as observed in HSkM cells.Copyright © 2017 the American Physiological Society.

Keyword: mitochondria

Dietary Olive and Perilla Oils Affect Liver Mitochondrial DNA Methylation in Large Yellow Croakers.

Substantial progress has been made in nutritional epigenetics, but little is known regarding whether mitochondrial DNA (mtDNA) methylation is involved in this process.The objective of this study was to determine whether dietary lipid sources [various fatty acids (FAs)] modify mtDNA methylation.A total of 600 large yellow croakers (Larimichthys crocea) with an average initial weight of 151 ± 4 g were fed 1 of 5 diets (3 replicate cages/treatment) containing either fish oil (FO) (control), , olive oil (OO), sunflower oil, or perilla oil (PO) as the dietary lipid source (12% dry weight of the diet) for 70 d. Pyrosequencing was used to determine the effects of dietary lipid sources (FAs) on mtDNA methylation.Mitochondrial arginine transfer RNA and NAD(H) dehydrogenase 4L encoding region methylation in the liver was higher in the OO (9.5% ± 0.52%; P < 0.05) and PO (7.3% ± 0.33%; P < 0.05) groups than in the FO (5.9% ± 0.42%) group, whereas 12S ribosomal RNA (rRNA) methylation in the liver was lower in the OO group (2.7% ± 0.22%) than in the FO group (4.2% ± 0.73%) (P < 0.05). Additionally, fish fed the OO diet had lower liver mRNA levels of ND3 (P < 0.05), ND4L (P < 0.05), ND6 (P < 0.05), 12S rRNA (P < 0.05), and 16S rRNA (P < 0.05) than those fed the FO diet, whereas fish fed the PO diet had lower liver mRNA levels of 16S rRNA than those fed the FO diet (P < 0.05). Moreover, fish fed the OO (P < 0.05) or PO (P < 0.05) diet had lower liver mitochondrial complex I activity than did those fed the FO diet.These findings provide the first evidence, to our knowledge, that dietary lipid sources influence mitochondrial function through mtDNA methylation in large yellow croakers.© 2015 American Society for Nutrition.

Keyword: mitochondria

The effect of chronic exposure to high concentrations on the aerobic metabolism of human endothelial EA.hy926 cells.

A chronic elevation of circulating free fatty acids (FFAs) is associated with diseases like obesity or diabetes and can lead to lipotoxicity. The goals of this study were to assess the influence of chronic exposure to high (PAL) levels on mitochondrial respiratory functions in endothelial cells and isolated . Human umbilical vein endothelial cells (EA.hy926 line) were grown for 6\xa0days in a medium containing either 100 or 150\xa0μM PAL. Growth at high PAL concentrations induced a considerable increase in fatty -supplied respiration and a reduction of mitochondrial respiration during carbohydrate and glutamine oxidation. High PAL levels elevated intracellular and mitochondrial superoxide generation; increased inflammation marker, acyl-coenzyme A (CoA) dehydrogenase, uncoupling protein 2 (UCP2), and superoxide dismutase 2 expression; and decreased hexokinase I and pyruvate dehydrogenase expression. No change in aerobic respiration capacity was observed, while fermentation was decreased. In isolated from high PAL-treated cells, an increase in the oxidation of palmitoylcarnitine, a decrease in the oxidation of pyruvate, and an increase in UCP2 activity were observed. Our results demonstrate that exposure to high PAL levels induces a shift in endothelial aerobic metabolism toward the oxidation of fatty acids. Increased levels of PAL caused impairment and uncoupling of the mitochondrial oxidative phosphorylation system. Our data indicate that FFAs significantly affect endothelial oxidative metabolism, reactive oxygen species (ROS) formation, and cell viability and, thus, might contribute to endothelial and vascular dysfunction.

Keyword: mitochondria

Peroxisomes can oxidize medium- and long-chain fatty acids through a pathway involving ABCD3 and HSD17B4.

Peroxisomes are essential organelles for the specialized oxidation of a wide variety of fatty acids, but they are also able to degrade fatty acids that are typically handled by mitochondria. Using a combination of pharmacological inhibition and clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 genome editing technology to simultaneously manipulate peroxisomal and mitochondrial fatty β-oxidation (FAO) in HEK-293 cells, we identified essential players in the metabolic crosstalk between these organelles. Depletion of carnitine palmitoyltransferase (CPT)2 activity through pharmacological inhibition or knockout (KO) uncovered a significant residual peroxisomal oxidation of lauric and , leading to the production of peroxisomal acylcarnitine intermediates. Generation and analysis of additional single- and double-KO cell lines revealed that the D-bifunctional protein (HSD17B4) and the peroxisomal ABC transporter ABCD3 are essential in peroxisomal oxidation of lauric and . Our results indicate that peroxisomes not only accept acyl-CoAs but can also oxidize acylcarnitines in a similar biochemical pathway. By using an Hsd17b4 KO mouse model, we demonstrated that peroxisomes contribute to the plasma acylcarnitine profile after acute inhibition of CPT2, proving in vivo relevance of this pathway. We summarize that peroxisomal FAO is important when mitochondrial FAO is defective or overloaded.-Violante, S., Achetib, N., van Roermund, C. W. T., Hagen, J., Dodatko, T., Vaz, F. M., Waterham, H. R., Chen, H., Baes, M., Yu, C., Argmann, C. A., Houten, S. M. Peroxisomes can oxidize medium- and long-chain fatty acids through a pathway involving ABCD3 and HSD17B4.

Keyword: mitochondria

Reduction of oxidative stress attenuates lipoapoptosis exacerbated by hypoxia in human hepatocytes.

Chronic intermittent hypoxia, a characteristic of obstructive sleep apnea (OSA), is associated with the progression of simple hepatic steatosis to necroinflammatory hepatitis. We determined whether inhibition of a hypoxia-induced signaling pathway could attenuate hypoxia-exacerbated lipoapoptosis in human hepatocytes. The human hepatocellular carcinoma cell line (HepG2) was used in this study. (PA)-treated groups were used for two environmental conditions: Hypoxia (1% O2) and normoxia (20% O2). Following the treatment, the cell viability was determined by the 3,4-(5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium salt (MTS) assay, and the mechanism of lipoapoptosis was evaluated by Western blotting. Hypoxia exacerbated the suppression of hepatocyte growth induced by via activation of mitochondrial apoptotic pathways as a result of endoplasmic reticulum (ER) and oxidative stresses. Ammonium pyrrolidine dithiocarbamate, a scavenger of reactive oxygen species, attenuated the hypoxia-exacerbated lipoapoptosis in hepatocytes, whereas glycerol, which reduces ER stress, did not. This may have been because inhibition of oxidative stress decreases the expression of pro-apoptotic proteins, such as caspase 9 and cytochrome c. These results suggested that modulation of apoptotic signaling pathways activated by oxidative stress can aid in identifying novel therapeutic strategies for the treatment of nonalcoholic steatohepatitis (NASH) with OSA. Further in vivo studies are necessary to understand the pathophysiologic mechanism of NASH with OSA and to prove the therapeutic effect of the modulation of the signaling pathways.

Keyword: mitochondria

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic -mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose . The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 diabetes.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: mitochondria

The beneficial effects of resveratrol on steatosis and mitochondrial oxidative stress in HepG2 cells.

Nonalcoholic fatty liver disease (NAFLD) is currently one of the most common chronic liver diseases, especially in developed countries. One group of substances with a potential use in the treatment of NAFLD are plant polyphenols, represented by resveratrol. The aim of this study was to evaluate the effect of resveratrol on steatosis and oxidative stress in HepG2 cells. The steatosis of cells was carried out using free fatty acids: oleic or and their mixtures. Steatosis was visualized using the intracellular lipid staining by Nile Red dye with a fluorescence microscope. This study also determined the viability of cells and mitochondrial membrane potential. The current study showed that fatty acids and their mixtures induced fat overloading in HepG2 cells. In the group of cells incubated with oleic (OA), observed changes were moderate with prevailing micro-vesicular steatosis. In case of cells incubated with (PA) and the mixtures of fatty acids, micro- and macro-vacuolar steatosis occurred in most of the cells. Resveratrol decreased steatosis in HepG2 cells induced by OA, PA, as well as their mixtures, and in most of experimental groups did not reduce cells viability. Resveratrol reduced the oxidative stress in HepG2 cells treated with fatty acids mixtures.

Keyword: mitochondria

Hydrogen Sulphide modulating mitochondrial morphology to promote mitophagy in endothelial cells under high-glucose and high-palmitate.

Endothelial cell dysfunction is one of the main reasons for type II diabetes vascular complications. Hydrogen sulphide (H S) has antioxidative effect, but its regulation on mitochondrial dynamics and mitophagy in aortic endothelial cells under hyperglycaemia and hyperlipidaemia is unclear. Rat aortic endothelial cells (RAECs) were treated with 40 mM glucose and 200 μM palmitate to imitate endothelium under hyperglycaemia and hyperlipidaemia, and 100 μM NaHS was used as an exogenous H S donor. Firstly, we demonstrated that high glucose and palmitate decreased H S production and CSE expression in RAECs. Then, the antioxidative effect of H S was proved in RAECs under high glucose and palmitate to reduce mitochondrial ROS level. We also showed that exogenous H S inhibited mitochondrial apoptosis in RAECs under high glucose and palmitate. Using Mito Tracker and transmission electron microscopy assay, we revealed that exogenous H S decreased mitochondrial fragments and significantly reduced the expression of p-Drp-1/Drp-1 and Fis1 compared to high-glucose and high-palmitate group, whereas it increased mitophagy by transmission electron microscopy assay. We demonstrated that exogenous H S facilitated Parkin recruited by PINK1 by immunoprecipitation and immunostaining assays and then ubiquitylated mitofusin 2 (Mfn2), which illuminated the mechanism of exogenous H S on mitophagy. Parkin siRNA suppressed the expression of Mfn2, Nix and LC3B, which revealed that it eliminated mitophagy. In summary, exogenous H S could protect RAECs against apoptosis under high glucose and palmitate by suppressing oxidative stress, decreasing mitochondrial fragments and promoting mitophagy. Based on these results, we proposed a new mechanism of H S on protecting endothelium, which might provide a new strategy for type II diabetes vascular complication.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: mitochondria

Effect of surface-potential modulators on the opening of lipid pores in liposomal and mitochondrial inner membranes induced by palmitate and calcium ions.

The effect of surface-potential modulators on palmitate/Ca2+-induced formation of lipid pores was studied in liposomal and inner mitochondrial membranes. Pore formation was monitored by sulforhodamine B release from liposomes and swelling of . ζ-potential in liposomes was determined from electrophoretic mobility. Replacement of sucrose as the osmotic agent with KCl decreased negative ζ-potential in liposomes and increased resistance of both and liposomes to the pore inducers, , and Ca2+. Micromolar Mg2+ also inhibited palmitate/Ca2+-induced permeabilization of liposomes. The rate of palmitate/Ca2+-induced, cyclosporin A-insensitive swelling of increased 22% upon increasing pH from 7.0 to 7.8. At below the critical micelle concentration, the cationic detergent cetyltrimethylammonium bromide (10 μM) and the anionic surfactant sodium dodecylsulfate (10-50 μM) made the ζ-potential less and more negative, respectively, and inhibited and stimulated opening of mitochondrial palmitate/Ca2+-induced lipid pores. Taken together, the findings indicate that surface potential regulates palmitate/Ca2+-induced lipid pore opening.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Proteomic effects of repeated-dose oral exposure to 2-monochloropropanediol and its dipalmitate in rat testes.

2- and 3-monochloropropanediol (2-MCPD) and their fatty esters are food contaminants which are concomitantly formed upon thermal treatment of foodstuff containing fats and salt. Exposure to 2- or 3-MCPD thus results, for example, from refined vegetable oils, in instant meals or infant formula, as well as in cereals or pastries. The molecular mechanisms of 2-MCPD toxicity are poorly understood. Here, we performed a comprehensive proteomic analysis of 2-MCDP-induced alterations in the testes from rats following oral administration of 10\u202fmg/kg body weight per day 2-MCPD, or an equimolar dose of 2-MCPD dipalmitate as a representative 2-MCPD fatty ester. In the absence of overt histopathologically detectable toxicity, moderate alterations in cellular proteomic signatures were recorded. The observations are in line with the assumption that the molecular mechanisms of 2-MCPD and 3-MCPD toxicity differ. Observed proteomic alterations point towards effects of 2-MCPD on mitogen-dependent signaling and mitochondrial energy utilization. Presented data for the first time provide insight into proteomic effects of 2-MCPD in testicular tissue.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Comparative study of changes in energy metabolism in rat cardiomyocytes in postinfarction cardiosclerosis and diabetes mellitus.

Cardiomyocyte energy metabolism in experimental unfolding postinfarction cardiosclerosis and diabetes mellitus was studied. Postinfarction cardiosclerosis formed 6 weeks after coronary artery occlusion. Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (60 mg/kg). The rate of oxygen consumption in postinfarction cardiosclerosis and diabetes increased by 3.4 and 4.2 times, respectively. Stimulation of mitochondrial respiration (ATP, ) significantly increased oxygen consumption in animals with postinfarction cardiosclerosis and significantly reduced this process in diabetes. The content of LDH and SDH in the myocardium of animals with diabetes and postinfarction cardiosclerosis was significantly below the control. Hence, the development of postinfarction cardiosclerosis and diabetes mellitus were characterized by reduced generation of ATP in anaerobic and aerobic pathways and oxidative phosphorylation in cardiomyocytes.

Keyword: mitochondria

High vulnerability of the heart and liver to 3-hydroxypalmitic -induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe liver and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic (3HPA), the long-chain hydroxyl fatty (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca retention capacity and membrane potential in Ca -loaded more markedly in the heart and the liver, with mild or no effects in the brain, supporting a higher susceptibility of the heart and the liver to the toxic effects of this fatty . It is postulated that disruption of mitochondrial energy and Ca homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.© 2018 Wiley Periodicals, Inc.

Keyword: mitochondria

Ca(2+)-dependent nonspecific permeability of the inner membrane of liver in the guinea fowl (Numida meleagris).

This comparative study presents the results of the induction of Ca(2+)-dependent nonspecific permeability of the inner membrane (pore opening) of rat and guinea fowl liver by mechanisms that are both sensitive and insensitive to cyclosporin A (CsA). It was established that energized rat and guinea fowl liver incubated with 1\xa0mM of inorganic phosphate (Pi) are capable of swelling upon addition of at least 125 and 875\xa0nmol of CaCl2 per 1\xa0mg protein, respectively. Under these conditions, the Ca(2+) release from the of these animals and a drop in Δψ are observed. All of these processes are inhibited by 1\xa0μM of CsA. FCCP, causing organelle de-energization, induces pore opening in rat and guinea fowl liver upon addition of 45 и 625\xa0nmol of CaCl2 per 1\xa0mg protein, respectively. These results suggest the existence of a CsA-sensitive mechanism for the induction of Ca(2+)-dependent pores in guinea fowl liver , which has been reported in rat liver . However, guinea fowl liver have a significantly greater resistance to Ca(2+) as a pore inducer compared to rat liver . It was found that the addition of α,ω-hexadecanedioic (HDA) to rat and guinea fowl liver incubated with CsA and loaded with Ca(2+) causes organelle swelling and Ca(2+) release from the matrix. It is assumed that in contrast to the CsA-sensitive pore, the CsA-insensitive pore induced by HDA in the inner membrane of guinea fowl liver , as well as in rat liver , is lipid in nature.

Keyword: mitochondria

Pterostilbene reverses mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 diabetes and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into . PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: mitochondria

Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles.

We developed a method that allows for real-time assessment of cellular metabolism in isolated, intact long skeletal muscle fibre bundles from adult mice. This method can be used to study changes in mitochondrial function and fuel utilisation in live skeletal muscle fibre bundles. Our method enables flexibility in experimental design and high-throughput assessment of mitochondrial parameters in isolated skeletal muscle fibre bundles. Extensor digitorum longus (EDL) fibre bundles obtained from chronic high-fat diet fed mice had lower basal oxygen consumption under FCCP-induced maximal respiration, when compared to control chow-fed mice. EDL fibre bundles obtained from chronic high-fat diet fed mice had enhanced mitochondrial oxidation capacity under FCCP-induced maximal respiration, when compared to control chow-fed mice.Metabolic dysfunction in skeletal muscle contributes to the aetiology and development of muscle diseases and metabolic diseases. As such, assessment of skeletal muscle cellular bioenergetics provides a powerful means to understand the role of skeletal muscle metabolism in disease and to identify possible therapeutic targets. Here, we developed a method that allows for the real-time assessment of cellular respiration in intact skeletal muscle fibre bundles obtained from the extensor digitorum longus (EDL) muscle of adult mice. Using this method, we assessed the contribution of ATP turnover and proton leak to basal mitochondrial oxygen consumption rate (OCR). Our data demonstrate that the in EDL fibres are loosely coupled. Moreover, in the presence of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), we show that palmitate exposure induced comparable peak OCR and higher total OCR in EDL fibre bundles when compared to pyruvate exposure, suggesting that fatty acids might be a more sustainable fuel source for skeletal muscle when are driven to maximal respiration. Application of this method to EDL fibre bundles obtained from chronic high-fat diet fed mice revealed lower basal OCR and enhanced mitochondrial oxidation capacity in the presence of FCCP when compared to the chow-diet fed control mice. By using a 96-well microplate format, our method provides a flexible and efficient platform to investigate mitochondrial parameters of intact skeletal muscle fibres obtained from adult mice.© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

Keyword: mitochondria

Hepatic toxicity of dronedarone in mice: role of mitochondrial β-oxidation.

Dronedarone is an amiodarone-like antiarrhythmic drug associated with severe liver injury. Since dronedarone inhibits mitochondrial respiration and β-oxidation in vitro, mitochondrial toxicity may also explain dronedarone-associated hepatotoxicity in vivo. We therefore studied hepatotoxicity of dronedarone (200mg/kg/day for 2 weeks or 400mg/kg/day for 1 week by intragastric gavage) in heterozygous juvenile visceral steatosis (jvs(+/-)) and wild-type mice. Jvs(+/-) mice have reduced carnitine stores and are sensitive for mitochondrial β-oxidation inhibitors. Treatment with dronedarone 200mg/kg/day had no effect on body weight, serum transaminases and bilirubin, and hepatic mitochondrial function in both wild-type and jvs(+/-) mice. In contrast, dronedarone 400mg/kg/day was associated with a 10-15% drop in body weight, and a 3-5-fold increase in transaminases and bilirubin in wild-type mice and, more accentuated, in jvs(+/-) mice. In vivo metabolism of intraperitoneal (14)C-palmitate was impaired in wild-type, and, more accentuated, in jvs(+/-) mice treated with 400mg/kg/day dronedarone compared to vehicle-treated mice. Impaired β-oxidation was also found in isolated ex vivo. A likely explanation for these findings was a reduced activity of carnitine palmitoyltransferase 1a in liver from dronedarone-treated mice. In contrast, dronedarone did not affect the activity of the respiratory chain ex vivo. We conclude that dronedarone inhibits mitochondrial β-oxidation in and ex vivo, but not the respiratory chain. Jvs(+/-) mice are slightly more sensitive for the effect of dronedarone on mitochondrial β-oxidation than wild-type mice. The results suggest that inhibition of mitochondrial β-oxidation is an important mechanism of hepatotoxicity associated with dronedarone.Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Keyword: mitochondria

Disturbance of mitochondrial functions provoked by the major long-chain 3-hydroxylated fatty acids accumulating in MTP and LCHAD deficiencies in skeletal muscle.

The pathogenesis of the muscular symptoms and recurrent rhabdomyolysis that are commonly manifested in patients with mitochondrial trifunctional protein (MTP) and long-chain 3-hydroxy-acyl-CoA dehydrogenase (LCHAD) deficiencies is still unknown. In this study we investigated the effects of the major long-chain monocarboxylic 3-hydroxylated fatty acids (LCHFA) accumulating in these disorders, namely 3-hydroxytetradecanoic (3HTA) and 3-hydroxypalmitic (3HPA) acids, on important mitochondrial functions in rat skeletal muscle . 3HTA and 3HPA markedly increased resting (state 4) and decreased ADP-stimulated (state 3) and CCCP-stimulated (uncoupled) respiration. 3HPA provoked similar effects in permeabilized skeletal muscle fibers, validating the results obtained in purified . Furthermore, 3HTA and 3HPA markedly diminished mitochondrial membrane potential, NAD(P)H content and Ca(2+) retention capacity in Ca(2+)-loaded . Mitochondrial permeability transition (mPT) induction probably underlie these effects since they were totally prevented by cyclosporin A and ADP. In contrast, the dicarboxylic analogue of 3HTA did not alter the tested parameters. Our data strongly indicate that 3HTA and 3HPA behave as metabolic inhibitors, uncouplers of oxidative phosphorylation and mPT inducers in skeletal muscle. It is proposed that these pathomechanisms disrupting mitochondrial homeostasis may be involved in the muscle alterations characteristic of MTP and LCHAD deficiencies.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Deregulation of mitochondrial functions provoked by long-chain fatty accumulating in long-chain 3-hydroxyacyl-CoA dehydrogenase and mitochondrial permeability transition deficiencies in rat heart--mitochondrial permeability transition pore opening as a potential contributing pathomechanism of cardiac alterations in these disorders.

Mitochondrial trifunctional protein and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies are fatty oxidation disorders biochemically characterized by tissue accumulation of long-chain fatty acids and derivatives, including the monocarboxylic long-chain 3-hydroxy fatty acids (LCHFAs) 3-hydroxytetradecanoic (3HTA) and 3-hydroxypalmitic (3HPA). Patients commonly present severe cardiomyopathy for which the pathogenesis is still poorly established. We investigated the effects of 3HTA and 3HPA, the major metabolites accumulating in these disorders, on important parameters of mitochondrial homeostasis in Ca(2+) -loaded heart . 3HTA and 3HPA significantly decreased mitochondrial membrane potential, the matrix NAD(P)H pool and Ca(2+) retention capacity, and also induced mitochondrial swelling. These fatty acids also provoked a marked decrease of ATP production reflecting severe energy dysfunction. Furthermore, 3HTA-induced mitochondrial alterations were completely prevented by the classical mitochondrial permeability transition (mPT) inhibitors cyclosporin A and ADP, as well as by ruthenium red, a Ca(2+) uptake blocker, indicating that LCHFAs induced Ca(2+)-dependent mPT pore opening. Milder effects only achieved at higher doses of LCHFAs were observed in brain , implying a higher vulnerability of heart to these fatty acids. By contrast, 3HTA and docosanoic acids did not change mitochondrial homeostasis, indicating selective effects for monocarboxylic LCHFAs. The present data indicate that the major LCHFAs accumulating in mitochondrial trifunctional protein and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies induce mPT pore opening, compromising Ca(2+) homeostasis and oxidative phosphorylation more intensely in the heart. It is proposed that these pathomechanisms may contribute at least in part to the severe cardiac alterations characteristic of patients affected by these diseases.© 2015 FEBS.

Keyword: mitochondria

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: mitochondria

[THE OPTIMIZATION OF NUTRITION FUNCTION UNDER SYNDROME OF RESISTANCE TO INSULIN, DISORDER OF FATTY ACIDS\' METABOLISM AND ABSORPTION OF GLUCOSE BY CELLS (A LECTURE)].

The phylogenetic processes continue to proceed in Homo Sapiens. At the very early stages ofphylogenesis, the ancient Archaea that formed under symbiotic interaction with later bacterial cells conjointly formed yet another system. In this system, there are no cells\' absorption of glucose if it is possible to absorb fatty acids from intercellular medium in the form of unesterfied fatty acids or ketonic bodies--metabolites of fatty acids. This is caused by objectively existed conditions and subsequent availability of substrates at the stages ofphylogenesis: acetate, ketonic bodies, fatty acids and only later glucose. The phylogenetically late insulin used after billions years the same dependencies at formation of regulation ofmetabolism offatty acids and cells\' absorption of glucose. In order that syndrome ofresistance ceased to exist as afoundation of metabolic pandemic Homo Sapiens has to understand the following. After successful function ofArchaea+bacterial cells and considered by biology action of insulin for the third time in phylogenesis and using biological function of intelligence the content ofphylogenetically earlier saturated fatty infood can\'t to exceed possibilities of phylogenetically late lipoproteins to transfer it in intercellular medium and blood and cells to absorb it. It is supposed that at early stages of phylogenesis biological function of intelligence is primarily formed to bring into line "unconformities" of regulation of metabolism against the background of seeming relative biological "perfection". These unconformities were subsequently and separately formed at the level of cells in paracrin regulated cenosises of cells and organs and at the level of organism. The prevention of resistance to insulin basically requires biological function of intelligence, principle of self-restraint, bringing into line multiple desires of Homo Sapiens with much less extensive biological possibilities. The "unconformities" of regulation of metabolism in vivo are etiological factors of all metabolic pandemics including atherosclerosis, metabolic arterial hypertension, obesity and metabolic syndrome Tertiannondatum.

Keyword: mitochondria

Azoramide improves mitochondrial dysfunction in palmitate-induced insulin resistant H9c2 cells.

Azoramide is identified as a new compound with the dual properties for the improvement of ER-folding capacity in various cells as well as for the treatment of T2DM. Although the effect of azoramide in glucose-homeostasis in mammalians is not known very well, a limited number of experimental studies showed that it could improve the insulin sensitivity in genetically obese mice. Therefore, here, we aimed to investigate the direct effect of azoramide on insulin signaling in insulin-resistant (IR) cardiomyocytes using IR-modelled ventricular cardiomyocytes. This model was established in H9c2 cells using incubation (50-μM for 24-h). The development of IR in cells was verified by monitoring the cellular 2-DG6P uptake assays in these treated cells. The 2-DG6P uptake was 50% less in the IR-cells compared to the control cells, while azoramide treatment (20-μM for 48-h) could prevent fully that decrease. In addition, azoramide treatment markedly preserved the IR-induced less ATP production and high-ROS production in these IR-cells. Furthermore, this treatment prevented the functional changes in mitochondria characterized by depolarized mitochondrial membrane potential and mitochondrial fusion or fusion-related protein levels as well as cellular ATP level. Moreover, this treatment provided marked protection against IR-associated changes in the insulin signaling pathway in cells, including recovery in the phosphorylation of IRS1 and Akt as well as the protein level of GLUT4 and Akt. Our present results, for the first time, demonstrated that azoramide plays an important protective role in IR-cardiomyocytes, at most, protective action on mitochondria. Therefore, one can suggest that azoramide, as a novel regulator, can provide direct cardioprotection in the IR-heart, at most, via affecting mitochondria and can be a good candidate as a new drug for the treatment of IR-associated cardiovascular disorders in mammalians with systemic IR.

Keyword: mitochondria

High-protein diets prevent steatosis and induce hepatic accumulation of monomethyl branched-chain fatty acids.

The hallmark of nonalcoholic fatty liver disease is steatosis of unknown etiology. To test how dietary protein decreases steatosis, we fed female C57BL/6 J mice low-fat (8 en%) or high-fat (42 en%) combined with low-protein (11 en%), high-protein (HP; 35 en%) or extra-high-protein (HPX; 58 en%) diets for 3 weeks. The 35 en% protein diets reduced hepatic triglyceride, free fatty , cholesterol and phospholipid contents to ~50% of that in 11 en% protein diets. Every additional 10 en% protein reduced hepatic fat content ~1.5 g%. HP diets had no effect on lipogenic or fatty -oxidizing genes except Ppargc1α (+30%), increased hepatic PCK1 content 3- to 5-fold, left plasma glucose and hepatic glycogen concentration unchanged, and decreased inflammation and cell stress (decreased Fgf21 and increased Gsta expression). The HP-mediated decrease in steatosis correlated inversely with plasma branched-chain amino- (BCAA) concentrations and hepatic content of BCAA-derived monomethyl branched-chain fatty acids (mmBCFAs) 14-methylpentadecanoic (14-MPDA; valine-derived) and, to a lesser extent, 14-methylhexadecanoic (isoleucine-derived). Liver lipid content was 1.6- to 1.8-fold higher in females than in males, but the anti-steatotic effect of HP diets was equally strong. The strong up-regulation of PCK1 and literature data showing an increase in phosphoenolpyruvate and a decline in tricarboxylic cycle intermediates in liver reveal that an increased efflux of these intermediates from represents an important effect of an HP diet. The HP diet-induced increase in 14-MPDA and the dietary response in gene expression were more pronounced in females than males. Our findings are compatible with a facilitating role of valine-derived mmBCFAs in the antisteatotic effect of HP diets.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: mitochondria

A permeability transition in liver and liposomes induced by α,ω-dioic acids and Ca(2+).

The article examines the molecular mechanism of the Ca(2+)-dependent cyclosporin A (CsA)-insensitive permeability transition in rat liver induced by α,ω-dioic acids. The addition of α,ω-hexadecanedioic (HDA) to Ca(2+)-loaded liver was shown to induce a high-amplitude swelling of the organelles, a drop of membrane potential and the release of Ca(2+) from the matrix, the effects being insensitive to CsA. The experiments with liposomes loaded with sulforhodamine B (SRB) revealed that, like (PA), HDA was able to cause permeabilization of liposomal membranes. However, the kinetics of HDA- and PA-induced release of SRB from liposomes was different, and HDA was less effective than PA in the induction of SRB release. Using the method of ultrasound interferometry, we also showed that the addition of Ca(2+) to HDA-containing liposomes did not change the phase state of liposomal membranes-in contrast to what was observed when Ca(2+) was added to PA-containing vesicles. It was suggested that HDA/Ca(2+)- and PA/Ca(2+)-induced permeability transition occurs by different mechanisms. Using the method of dynamic light scattering, we further revealed that the addition of Ca(2+) to HDA-containing liposomes induced their aggregation/fusion. Apparently, these processes result in a partial release of SRB due to the formation of fusion pores. The possibility that this mechanism underlies the HDA/Ca(2+)-induced permeability transition of the mitochondrial membrane is discussed.

Keyword: mitochondria

H2O2 release from the very long chain acyl-CoA dehydrogenase.

Enhanced mitochondrial generation of oxidants, including hydrogen peroxide (H2O2), is related to a large number of pathological conditions, including diet-induced obesity and steatohepatosis. Indeed, we have previously shown that high fat diets increase the generation of H2O2 in liver energized by activated fatty acids. Here, we further study fatty- induced H2O2 release in liver , and determine the characteristics that regulate it. We find that this production of H2O2 is independent of mitochondrial inner membrane integrity and insensitive to purine nucleotides. On the other hand, palmitate-induced H2O2 production is strongly enhanced by high fat diets and is pH-sensitive, with a peak at a matrix pH of ~8.5. Using recombinantly expressed human very long chain acyl-CoA dehydrogenase, we are able to demonstrate that palmitate-induced H2O2 release may be ascribed to the activity of this enzyme alone, acting as an oxidase. Our results add to a number of findings indicating that sources outside of the electron transport chain can generate significant, physiopathologically relevant, amounts of oxidants in .Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Multiple pathways are involved in -induced toxicity.

In this study, we identified the toxic mechanism following the accumulation of (PA), a saturated fatty , in human Chang liver cells. After PA exposure for 24 h, the and the endoplasmic reticulum (ER) became dilated, and lipid droplets and organelles were observed within autophagosomes. Cell viability decreased with an ATP reduction and the G2/M phase arrest. The expression of SOD-2, but not of SOD-1, markedly increased after PA exposure, which also elevated the number of cells generating ROS. PA enhanced the levels of proteins related to apoptosis, necroptosis, autophagy, and ER stress. Moreover, the inhibition of caspases, p53, necroptosis, or ER stress substantially rescued PA-induced cytotoxicity and, similarly, the inhibition of caspases and ER stress counteracted PA-induced changes in the cell cycle. Conversely, the inhibition of necroptosis and p53 signaling accelerated the changes in the cell cycle triggered by PA exposure. Blocking autophagy exacerbated PA-induced cytotoxicity and alterations in the cell cycle and caused disappearance of cellular components. These results suggest that PA induces apoptosis accompanied by autophagy through mitochondrial dysfunction and ER stress, which are triggered by oxidative stress in Chang liver cells and that blocking autophagy accelerates cell damage following PA exposure.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Fatty chain elongation in palmitate-perfused working rat heart: mitochondrial acetyl-CoA is the source of two-carbon units for chain elongation.

Rat hearts were perfused with [1,2,3,4-(13)C4] (M+4), and the isotopic patterns of myocardial acylcarnitines and acyl-CoAs were analyzed using ultra-HPLC-MS/MS. The 91.2% (13)C enrichment in palmitoylcarnitine shows that little endogenous (M+0) palmitate contributed to its formation. The presence of M+2 myristoylcarnitine (95.7%) and M+2 acetylcarnitine (19.4%) is evidence for β-oxidation of perfused M+4 . Identical enrichment data were obtained in the respective acyl-CoAs. The relative (13)C enrichment in M+4 (84.7%, 69.9%) and M+6 (16.2%, 17.8%) stearoyl- and arachidylcarnitine, respectively, clearly shows that the perfused palmitate is chain-elongated. The observed enrichment of (13)C in acetylcarnitine (19%), M+6 stearoylcarnitine (16.2%), and M+6 arachidylcarnitine (17.8%) suggests that the majority of two-carbon units for chain elongation are derived from β-oxidation of [1,2,3,4-(13)C4]. These data are explained by conversion of the M+2 acetyl-CoA to M+2 malonyl-CoA, which serves as the acceptor for M+4 palmitoyl-CoA in chain elongation. Indeed, the (13)C enrichment in mitochondrial acetyl-CoA (18.9%) and malonyl-CoA (19.9%) are identical. No (13)C enrichment was found in acylcarnitine species with carbon chain lengths between 4 and 12, arguing against the simple reversal of fatty β-oxidation. Furthermore, isolated, intact rat heart 1) synthesize malonyl-CoA with simultaneous inhibition of carnitine palmitoyltransferase 1b and 2) catalyze the palmitoyl-CoA-dependent incorporation of (14)C from [2-(14)C]malonyl-CoA into lipid-soluble products. In conclusion, rat heart has the capability to chain-elongate fatty acids using -derived two-carbon chain extenders. The data suggest that the chain elongation process is localized on the outer surface of the mitochondrial outer membrane.

Keyword: mitochondria

miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: mitochondria

Tibolone Preserves Mitochondrial Functionality and Cell Morphology in Astrocytic Cells Treated with .

Obesity has been associated with increased chronic neuroinflammation and augmented risk of neurodegeneration. This is worsened during the normal aging process when the levels of endogenous gonadal hormones are reduced. In this study, we have assessed the protective actions of tibolone, a synthetic steroid with estrogenic actions, on T98G human astrocytic cells exposed to , a saturated fatty used to mimic obesity in vitro. Tibolone improved cell survival, and preserved mitochondrial membrane potential in -treated astrocytic cells. Although we did not find significant actions of tibolone on free radical production, it modulated astrocytic morphology after treatment with . These data suggest that tibolone protects astrocytic cells by preserving both mitochondrial functionality and morphological complexity.

Keyword: mitochondria

Sirtuin 5 overexpression attenuates glucolipotoxicity-induced pancreatic β cells apoptosis and dysfunction.

Recently, SIRT5 was reported to be a predominant desuccinylase and demalonylase in . Ablation of SIRT5 enhances the systemic succinylation and malonylation of mitochondrial proteins, including various metabolic enzymes; however, its function in pancreatic β cells has not yet been clarified. In this study, we evaluated the effects of SIRT5 overexpression on glucolipotoxicity-induced apoptosis in β cell lines. Full-length SIRT5, which preferentially targeted to and partially to the nucleus and cytoplasm, was overexpressed in NIT-1 cells. Chronic exposure to excess palmitate and glucose (High-PA-G) induced apoptosis and suppressed glucose-stimulated insulin secretion in β cells. SIRT5 overexpression significantly alleviated apoptosis under the High-PA-G condition, accompanied by suppressed Caspase 3 activity and reduced malondialdehyde levels. SIRT5 overexpression also improved β cell secretory capacity in response to glucose under the High-PA-G condition, suggesting its protective role in β cell function. Furthermore, SIRT5 overexpression reversed the decreasing trend of anti-apoptotic factors BCL-2 and BCL-XL expression under High-PA-G condition. Further regulation mechanisms between SIRT5 and these anti-apoptotic factors remains to be explored in future studies. Our data reveal that SIRT5 is a potentially protective factor for pancreatic β cells against glucolipotoxicity-induced apoptosis and cell dysfunction.Copyright © 2018. Published by Elsevier Inc.

Keyword: mitochondria

Sab (Sh3bp5) dependence of JNK mediated inhibition of mitochondrial respiration in induced hepatocyte lipotoxicity.

Sustained c-Jun N-terminal kinase (JNK) activation by saturated fatty acids plays a role in lipotoxicity and the pathogenesis of non-alcoholic steatohepatitis (NASH). We have reported that the interaction of JNK with mitochondrial Sab leads to inhibition of respiration, increased reactive oxygen species (ROS), cell death and hepatotoxicity. We tested whether this pathway underlies (PA)-induced lipotoxicity in hepatocytes.Primary mouse hepatocytes (PMH) from adeno-shlacZ or adeno-shSab treated mice and HuH7 cells were used.In PMH, PA dose-dependently up to 1mM stimulated oxygen consumption rate (OCR) due to mitochondrial β-oxidation. At ⩾1.5mM, PA gradually reduced OCR, followed by cell death. Inhibition of JNK, caspases or treatment with antioxidant butylated hydroxyanisole (BHA) protected PMH against cell death. Sab knockdown or a membrane permeable Sab blocking peptide prevented PA-induced mitochondrial impairment, but inhibited only the late phase of both JNK activation (beyond 4h) and cell death. In PMH, PA increased p-PERK and its downstream target CHOP, but failed to activate the IRE-1α arm of the UPR. However, Sab silencing did not affect PA-induced PERK activation. Conversely, specific inhibition of PERK prevented JNK activation and cell death, indicating a major role upstream of JNK activation.The effect of p-JNK on plays a key role in PA-mediated lipotoxicity. The interplay of p-JNK with mitochondrial Sab leads to impaired respiration, ROS production, sustained JNK activation, and apoptosis.Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Dipeptidyl peptidase 9 enzymatic activity influences the expression of neonatal metabolic genes.

The success of dipeptidyl peptidase 4 (DPP4) inhibition as a type 2 diabetes therapy has encouraged deeper examination of the post-proline DPP enzymes. DPP9 has been implicated in immunoregulation, disease pathogenesis and metabolism. The DPP9 enzyme-inactive (Dpp9 gene knock-in; Dpp9 gki) mouse displays neonatal lethality, suggesting that DPP9 enzyme activity is essential in neonatal development. Here we present gene expression patterns in these Dpp9 gki neonatal mice. Taqman PCR arrays and sequential qPCR assays on neonatal liver and gut revealed differential expression of genes involved in cell growth, innate immunity and metabolic pathways including long-chain-fatty- uptake and esterification, long-chain fatty acyl-CoA binding, trafficking and transport into , lipoprotein metabolism, adipokine transport and gluconeogenesis in the Dpp9 gki mice compared to wild type. In a liver cell line, Dpp9 knockdown increased AMP-activated protein kinase phosphorylation, which suggests a potential mechanism. DPP9 protein levels in liver cells were altered by treatment with EGF, HGF, insulin or palmitate, suggesting potential natural DPP9 regulators. These gene expression analyses of a mouse strain deficient in DPP9 enzyme activity show, for the first time, that DPP9 enzyme activity regulates metabolic pathways in neonatal liver and gut.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: mitochondria

3,5-diiodo-L-thyronine modifies the lipid droplet composition in a model of hepatosteatosis.

Fatty acids are the main energy stores and the major membrane components of the cells. In the hepatocyte, fatty acids are esterified to triacylglycerols (TAGs) and stored in lipid droplets (LDs). The lipid lowering action of 3,5-diiodo-L-thyronine (T2) on an in vitro model of hepatosteatosis was investigated in terms of fatty and protein content of LDs, lipid oxidation and secretion.FaO cells were exposed to oleate/palmitate, then treated with T2.T2 reduced number and size of LDs, and modified their acyl composition by decreasing the content of saturated (SFA) vs monounsaturated (MUFA) fatty acids thus reversing the SFA/MUFA ratio. The expression of the LD-associated proteins adipose differentiation-related protein (ADRP), oxidative tissue-enriched PAT protein (OXPAT), and adipose triglyceride lipase (ATGL) was increased in \'steatotic\' cells and further up-regulated by T2. Moreover, T2 stimulated the mitochondrial oxidation by up-regulating carnitine-palmitoyl-transferase (CPT1), uncoupling protein 2 (UCP2) and very long-chain acyl-coenzyme A dehydrogenase (VLCAD).T2 leads to mobilization of TAGs from LDs and stimulates mitochondrial oxidative metabolism of fatty acids, in particular of SFAs, and thus enriches of MUFAs the LDs. This action may protect the hepatocyte from excess of SFAs that are more toxic than MUFAs.© 2014 S. Karger AG, Basel.

Keyword: mitochondria

Maternal overnutrition by hypercaloric diets programs hypothalamic mitochondrial fusion and metabolic dysfunction in rat male offspring.

Maternal overnutrition including pre-pregnancy, pregnancy and lactation promotes a lipotoxic insult leading to metabolic dysfunction in offspring. Diet-induced obesity models (DIO) show that changes in hypothalamic fusion and fission dynamics modulate metabolic dysfunction. Using three selective diet formula including a High fat diet (HFD), Cafeteria (CAF) and High Sugar Diet (HSD), we hypothesized that maternal diets exposure program leads to selective changes in hypothalamic fusion and fission dynamics in male offspring leading to metabolic dysfunction which is exacerbated by a second exposure after weaning.We exposed female Wistar rats to nutritional programming including Chow, HFD, CAF, or HSD for 9\xa0weeks (pre-mating, mating, pregnancy and lactation) or to the same diets to offspring after weaning. We determined body weight, food intake and metabolic parameters in the offspring from 21 to 60\xa0days old. Hypothalamus was dissected at 60\xa0days old to determine -ER interaction markers by mRNA expression and western blot and morphology by transmission electron microscopy (TEM). Mitochondrial-ER function was analyzed by confocal microscopy using hypothalamic cell line mHypoA-CLU192.Maternal programming by HFD and CAF leads to failure in glucose, leptin and insulin sensitivity and fat accumulation. Additionally, HFD and CAF programming promote mitochondrial fusion by increasing the expression of MFN2 and decreasing DRP1, respectively. Further, TEM analysis confirms that CAF exposure after programing leads to an increase in fusion and enhanced mitochondrial-ER interaction, which partially correlates with metabolic dysfunction and fat accumulation in the HFD and CAF groups. Finally, we identified that lipotoxic stimulus in hypothalamic cells increases Ca overload into matrix leading to mitochondrial dysfunction.We concluded that maternal programming by HFD induces hypothalamic fusion, metabolic dysfunction and fat accumulation in male offspring, which is exacerbated by HFD or CAF exposure after weaning, potentially due to calcium overflux.

Keyword: mitochondria

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial .A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: mitochondria

BNIP3 induction by hypoxia stimulates FASN-dependent free fatty production enhancing therapeutic potential of umbilical cord blood-derived human mesenchymal stem cells.

Mitophagy under hypoxia is an important factor for maintaining and regulating stem cell functions. We previously demonstrated that fatty synthase (FASN) induced by hypoxia is a critical lipid metabolic factor determining the therapeutic efficacy of umbilical cord blood-derived human mesenchymal stem cells (UCB-hMSCs). Therefore, we investigated the mechanism of a major mitophagy regulator controlling lipid metabolism and therapeutic potential of UCB-hMSCs. This study revealed that Bcl2/adenovirus E1B 19kDa protein-interacting protein 3 (BNIP3)-dependent mitophagy is important for reducing mitochondrial reactive oxygen species accumulation, anti-apoptosis, and migration under hypoxia. And, BNIP3 expression was regulated by CREB binding protein-mediated transcriptional actions of HIF-1α and FOXO3. Silencing of BNIP3 suppressed free fatty (FFA) synthesis regulated by SREBP1/FASN pathway, which is involved in UCB-hMSC apoptosis via caspases cleavage and migration via cofilin-1-mediated F-actin reorganization in hypoxia. Moreover, reduced mouse skin wound-healing capacity of UCB-hMSC with hypoxia pretreatment by BNIP3 silencing was recovered by . Collectively, our findings suggest that BNIP3-mediated mitophagy under hypoxia leads to FASN-induced FFA synthesis, which is critical for therapeutic potential of UCB-hMSCs with hypoxia pretreatment.Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Sonodynamic therapy inhibits palmitate-induced beta cell dysfunction via PINK1/Parkin-dependent mitophagy.

In type 2 diabetes mellitus (T2DM), the overload of glucose and lipids can promote oxidative stress and inflammatory responses and contribute to the failure of beta cells. However, therapies that can modulate the function of beta cells and thus prevent their failure have not been well explored. In this study, beta cell injury model was established with (PA) to simulate the lipotoxicity (high-fat diet) found in T2DM. Sonodynamic therapy (SDT), a novel physicochemical treatment, was applied to treat injured beta cells. We found that SDT had specific effects on and induced transient large amount of mitochondrial reactive oxygen species (ROS) production in beta cells. SDT also improved the morphology and function of abnormal , inhibited inflammatory response and reduced beta cell dysfunction. The improvement of was mediated by PINK1/Parkin-dependent mitophagy. Additionally, SDT rescued the transcription of PINK1 mRNA which was blocked by PA treatment, thus providing abundant PINK1 for mitophagy. Moreover, SDT also increased insulin secretion from beta cells. The protective effects of SDT were abrogated when mitophagy was inhibited by cyclosporin A (CsA). In summary, SDT potently inhibits lipotoxicity-induced beta cell failure via PINK1/Parkin-dependent mitophagy, providing theoretical guidance for T2DM treatment in aspects of islet protection.

Keyword: mitochondria

Theaflavins Improve Insulin Sensitivity through Regulating Mitochondrial Biosynthesis in -Induced HepG2 Cells.

Theaflavins, the characteristic and bioactive polyphenols in black tea, possess the potential improving effects on insulin resistance-associated metabolic abnormalities, including obesity and type 2 diabetes mellitus. However, the related molecular mechanisms are still unclear. In this research, we investigated the protective effects of theaflavins against insulin resistance in HepG2 cells induced by . Theaflavins significantly increased glucose uptake of insulin-resistant cells at noncytotoxic doses. This activity was mediated by upregulating the total and membrane bound glucose transporter 4 protein expressions, increasing the phosphor-Akt (Ser473) level, and decreasing the phosphorylation of IRS-1 at Ser307. Moreover, theaflavins were found to enhance the mitochondrial DNA copy number, down-regulate the PGC-1β mRNA level and increase the PRC mRNA expression. Mdivi-1, a selective mitochondrial division inhibitor, could attenuate TFs-induced promotion of glucose uptake in insulin-resistant HepG2 cells. Taken together, these results suggested that theaflavins could improve hepatocellular insulin resistance induced by free fatty acids, at least partly through promoting mitochondrial biogenesis. Theaflavins are promising functional food ingredients and medicines for improving insulin resistance-related disorders.

Keyword: mitochondria

H₂O₂-Activated Mitochondrial Phospholipase iPLA₂γ Prevents Lipotoxic Oxidative Stress in Synergy with UCP2, Amplifies Signaling via G-Protein-Coupled Receptor GPR40, and Regulates Insulin Secretion in Pancreatic β-Cells.

Pancreatic β-cell chronic lipotoxicity evolves from acute free fatty (FA)-mediated oxidative stress, unprotected by antioxidant mechanisms. Since mitochondrial uncoupling protein-2 (UCP2) plays antioxidant and insulin-regulating roles in pancreatic β-cells, we tested our hypothesis, that UCP2-mediated uncoupling attenuating mitochondrial superoxide production is initiated by FA release due to a direct H2O2-induced activation of mitochondrial phospholipase iPLA2γ.Pro-oxidant tert-butylhydroperoxide increased respiration, decreased membrane potential and mitochondrial matrix superoxide release rates of control but not UCP2- or iPLA2γ-silenced INS-1E cells. iPLA2γ/UCP2-mediated uncoupling was alternatively activated by an H2O2 burst, resulting from (PA) β-oxidation, and it was prevented by antioxidants or catalase overexpression. Exclusively, nascent FAs that cleaved off phospholipids by iPLA2γ were capable of activating UCP2, indicating that the previously reported direct redox UCP2 activation is actually indirect. Glucose-stimulated insulin release was not affected by UCP2 or iPLA2γ silencing, unless pro-oxidant activation had taken place. PA augmented insulin secretion via G-protein-coupled receptor 40 (GPR40), stimulated by iPLA2γ-cleaved FAs (absent after GPR40 silencing).The iPLA2γ/UCP2 synergy provides a feedback antioxidant mechanism preventing oxidative stress by physiological FA intake in pancreatic β-cells, regulating glucose-, FA-, and redox-stimulated insulin secretion. iPLA2γ is regulated by exogenous FA via β-oxidation causing H2O2 signaling, while FAs are cleaved off phospholipids, subsequently acting as amplifying messengers for GPR40. Hence, iPLA2γ acts in eminent physiological redox signaling, the impairment of which results in the lack of antilipotoxic defense and contributes to chronic lipotoxicity.

Keyword: mitochondria

Analysis of Titin in Red and White Muscles: Crucial Role on Muscle Contractions Using a Fish Model.

Several studies have compared molecular components between red and white skeletal muscles in mammals. However, mammalian skeletal muscles are composed of mixed types of muscle fibers. In the current study, we analyzed and compared the distributions of titin, lipid, phosphate ions, and fatty levels in red and white muscles using a fish model (), which is rich in red and white muscles, and these are well separated. Oil-red O staining showed that red muscle had more-abundant lipids than did white muscle. A time-of-flight secondary-ion mass spectrometric (TOF-SIMS) analysis revealed that red muscle possessed high levels of and oleic , but white muscle contained more phosphate ions. Moreover, elastica-van Gieson (EVG) and Mito-Tracker green FM staining showed that collagen and elastic fibers were highly, respectively, distributed in connective tissues and in red muscle. An electron micrographic analysis indicated that red muscle had a relatively higher number of and longer sarcomere lengths and Z-line widths, while myofibril diameters were thicker in white muscle. Myofibrillar proteins separated by SDS-PAGE showed that the major giant protein, titin, was highly expressed in white muscle than in red muscle. Furthermore, ratios of titin to myosin heavy chain (MHC) (titin/MHC) were about 1.3 times higher in white muscle than red muscle. We postulated that white muscle is fit for short and strong contractile performance due to high levels of titin and condensed sarcomeres, whereas red muscle is fit for low intensity and long-lasting activity due to high levels of lipids and and long sarcomeres.

Keyword: mitochondria

Protocatechuic Ameliorated -Induced Oxidative Damage in Endothelial Cells through Activating Endogenous Antioxidant Enzymes via an Adenosine-Monophosphate-Activated-Protein-Kinase-Dependent Pathway.

Protocatechuic (PCA, 3,4-dihydroxybenzoic ), the main metabolite of anthocyanins, is widely distributed in fruits and vegetables and has been reported to possess a strong antioxidant activity. Herein, we aimed to investigate the protective effect of PCA against high (PA)-induced oxidative damage and the underling molecular mechanisms in human umbilical vein endothelial cells (HUVECs). PCA reduced the levels of intracellular reactive oxygen species and malondialdehyde and increased the activities of endogenous antioxidant enzymes, including superoxide dismutase, glutathione peroxidase 1, and heme oxygenase 1 (HO-1). Metabolomic analysis showed that PCA affected numerous metabolites, especially some of which were related with energy . PCA also upregulated the phosphorylation of adenosine-monophosphate-activated protein kinase (AMPK) at Thr through activating liver kinase B1 and then promoted the expression of p-Nrf2 and HO-1. Moreover, PCA reversed the decreased expression of peroxisome proliferator-activated receptor γ coactivator 1α and significantly increased the mitochondrial density. Collectively, these results demonstrated that PCA attenuated PA-induced oxidative damage in HUVECs via an AMPK-dependent pathway.

Keyword: mitochondria

In Vitro Monitoring of the Mitochondrial Beta-Oxidation Flux of and Investigation of Its Pharmacological Alteration by Therapeutics.

The present study was designed to validate the functional assay that enables rapid screening of therapeutic candidates for their effect on mitochondrial fatty oxidation.The two whole-cell systems (tissue homogenates and hepatocytes) have been evaluated to monitor the total beta-oxidation flux of physiologically important H- by measurement of tritiated water enrichment in incubations using UPLC coupled on-line to radioactivity monitoring and mass spectrometry.Our results with several known inhibitors of fatty oxidation showed that this simple assay could correctly predict a potential in alteration of mitochondrial function by drug candidates. Since the beta-oxidation of takes place almost exclusively in of human hepatocytes, this model can be also utilized to distinguish between the mitochondrial and peroxisomal routes of this essential metabolic pathway in some cases.The present work offers a new in vitro screen of changes in mitochondrial beta-oxidation by xenobiotics as well as a model to study the mechanism of this pathway.

Keyword: mitochondria

[Influence of Fatty Acids on Oxygen Consumption in Isolated Cardiomyocytes of Rats with Ischemic or Diabetic Heart Disease].

one of the reasons of violation of the functional viability of the myocardium is considered to be the oxygen deprivation and lack of energy. The reason is the inhibitory effect of fatty acids on glucose oxidation. Recently, however, new data have been published proving the need for fatty acids and their importance in the maintenance and regulation of the functional activity of the myocardium in chronic pathology.to investigate the influence of free polyunsaturated and saturated fatty acids (FA) on the oxygen uptake of isolated cardiomyocytes in intact rats and animals with ischemic or diabetic heart disease.the executed non-randomized controlled study. It includied 3 groups of male rats of Wistar line (weight 250-300g) with 10 animals in each group. Myocardial infarction ("heart attack" group) was caused by ligation of the left coronary artery, diabetes ("diabetes" group)--by intraperitoneal injection of streptozotocin, and "control" group (intact animals). Myocardial infarction caused by ligation of the left coronary artery, and diabetes by intraperitoneal injection of streptozotocin. Isolated cardiac myocytes were obtained by the enzymatic method. Oxygen consumption was assessed polarographically at different saturation incubation medium with oxygen ([O₂] ≤ 8 mg/l and ([O₂] ≥ 16 mg/l). Arachidonic and acids were applied as fatty acids.It is established that the introduction of the incubation medium 20 µm arachidonic or fatty significantly increased the oxygen consumption of intact cardiomyocytes of rats. Both at the ischemic and at the diabetic injury to the heart the opposite result was obtained. The most pronounced decrease in oxygen consumption was indicated in the group with diabetes mellitus.The inhibitory effect of LCD on the rate of oxygen consumption may be associated with the influence of the ischemic or diabetic injury to the heart on the barrierfunction ofmitochondrial membranes of cardiomyocytes, the activity of membrane-associated enzymes and their associated processes.

Keyword: mitochondria

A component of the mitochondrial outer membrane proteome of T. brucei probably contains covalent bound fatty acids.

A subclass of eukaryotic proteins is subject to modification with fatty acids, the most common of which are and myristic . Protein acylation allows association with cellular membranes in the absence of transmembrane domains. Here we examine POMP39, a protein previously described to be present in the outer mitochondrial membrane proteome (POMP) of the protozoan parasite Trypanosoma brucei. POMP39 lacks canonical transmembrane domains, but is likely both myristoylated and palmitoylated on its N-terminus. Interestingly, the protein is also dually localized on the surface of the mitochondrion as well as in the flagellum of both insect-stage and the bloodstream form of the parasites. Upon abolishing of global protein acylation or mutation of the myristoylation site, POMP39 relocates to the cytosol. RNAi-mediated ablation of the protein neither causes a growth phenotype in insect-stage nor bloodstream form trypanosomes.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: mitochondria

The inhibition of Nrf2 accelerates renal lipid deposition through suppressing the ACSL1 expression in obesity-related nephropathy.

Obesity has become a worldwide epidemic, and the incidence of obesity is increasing year by year. Obesity-related nephropathy (ORN) is a common kidney complication of obesity. Long-chain acyl-CoA synthetases-1, (ACSL1), is a key enzyme in the oxidative metabolism of fatty acids in and ACSL1 may play a direct role in renal lipid deposition and promote the progress of ORN. In this study, we focus on the renoprotective role of ACSL1 in ORN. Electron microscopy, immunohistochemical (IHC) staining, Western blot, and real-time PCR were used to detect the expression of ACSL1and Nrf2 in ORN patients, ob/ob mice and (PA)-treated HK-2 cells. Oil red staining and Elisa Kit were used to detect the intracellular FFA and TG contents in ob/ob mice and PA-treated HK-2 cells. Dihydroethidium (DHE) staining and the MDA/SOD measurement were used to detect the ROS production. In order to demonstrate the role of ACSL1 and the interaction between ACSL1 and Nrf2 in ORN, related siRNA and plasmid were transfected into HK-2 cells. More ROS production and renal lipid deposition have been found in ORN patients, ob/ob mice and PA-treated HK-2 cells. Compared with control, all the expression of ACSL1and Nrf2 were down-regulated in ORN patients, ob/ob mice and PA-treated HK-2 cells. The Nrf2 could regulate the expression of ACSL1 and the ACSL1 played the direct role in renal lipid deposition. The Nrf2 is inhibited in ORN, resulting more ROS production and oxidative stress. Increased oxidative stress will suppress the expression of ACSL1, which could increase the intracellular FFA and TG contents, ultimately leading to renal lipid deposition in renal tubulars and accelerating the development of ORN.

Keyword: mitochondria

Mitochondrial bioenergetics deregulation caused by long-chain 3-hydroxy fatty acids accumulating in LCHAD and MTP deficiencies in rat brain: a possible role of mPTP opening as a pathomechanism in these disorders?

Long-chain 3-hydroxylated fatty acids (LCHFA) accumulate in long-chain 3-hydroxy-acyl-CoA dehydrogenase (LCHAD) and mitochondrial trifunctional protein (MTP) deficiencies. Affected patients usually present severe neonatal symptoms involving cardiac and hepatic functions, although long-term neurological abnormalities are also commonly observed. Since the underlying mechanisms of brain damage are practically unknown and have not been properly investigated, we studied the effects of LCHFA on important parameters of mitochondrial homeostasis in isolated from cerebral cortex of developing rats. 3-Hydroxytetradecanoic (3 HTA) reduced mitochondrial membrane potential, NAD(P)H levels, Ca(2+) retention capacity and ATP content, besides inducing swelling, cytochrome c release and H2O2 production in Ca(2+)-loaded mitochondrial preparations. We also found that cyclosporine A plus ADP, as well as ruthenium red, a Ca(2+) uptake blocker, prevented these effects, suggesting the involvement of the mitochondrial permeability transition pore (mPTP) and an important role for Ca(2+), respectively. 3-Hydroxydodecanoic and 3-hydroxypalmitic acids, that also accumulate in LCHAD and MTP deficiencies, similarly induced mitochondrial swelling and decreased ATP content, but to a variable degree pending on the size of their carbon chain. It is proposed that mPTP opening induced by LCHFA disrupts brain bioenergetics and may contribute at least partly to explain the neurologic dysfunction observed in patients affected by LCHAD and MTP deficiencies.Copyright © 2014 Elsevier B.V. All rights reserved.

Keyword: mitochondria

[The physical chemical and biological features of triglycerides. The cell absorption of functionally different +oleic lipoproteins of very low and density and linoleic+linolenic lipoproteins of low density.]

The earlier insulin-independent low-density lipoproteins and more late insulin-dependent very low-density lipoproteins implement different functions at the stages of phylogenesis. The disorder of biological function of trophology, alteration of fatty acids in triglycerides, prevalence of very low-density lipoproteins over oleic very low-density lipoproteins supply of cells with non-optimal substrate - saturated fatty for gaining energy, ATP synthesis. Physiologically, cells implement oleic alternative of fatty acids metabolism, oxidizing mainly ω-9 endogenous oleic mono-unsaturated fatty . The pathology of low density lipoproteins is primary deficiency of poly-unsaturated fatty acids in cells, atherosclerosis and atheromotosis of intima of arteries of elastic type with development of dense plaques from poly-unsaturated fatty acids in the form of polyethers of cholesterol. The pathology of very low-density lipoproteins includes: a) syndrome of resistance to insulin; b) pathology of phylogenetically earlier insulin-independent visceral fatty tissue - metabolic syndrome; c) pathology of phylogenetically later insulin-dependent subcutaneous adipocytes - obesity; d) secondary atherosclerosis, under cumulation of low-density lipoproteins in blood with development of atherothrombosis of intima of arteries, soft plaques rich with triglycerides. As for the prevention of disorders of transfer of fatty acids to very low-density lipoproteins and low-density lipoproteins is common in many ways - minimization of aphysiological effect of surplus amount of food, biological function of diet. The prevention at the level of population includes: a) maximal limitation of content of saturated fatty in food; b) moderate increasing of polysaturated fatty acids, ω-3 poly-saturated fatty acids predominantly; c) increasing of physical activity. The pharmaceuticals are not provided by biology in primary prevention of metabolic pandemics under aphysiological impact of environment factors.

Keyword: mitochondria

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize ; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: mitochondria

[ induces hepatocellular oxidative stress and activation of inflammasomes].

To evaluate the effect of (PA) on oxidative stress and activation of inflammasomes in hepatocytes.To test the dose-dependent effect of PA on normal murine hepatocytes AML12, the cells were treated with 0, 0.15, 0.25 and 0.4 mmol/L of (PA). The cells were also divided into blank control group, 0.25 mmol/L PA group and 0.25 mmol/L PA+N-acetylcysteine (NAC) group to examine the effect of reactive oxygen species (ROS) on the activation of inflammasomes. After 24 h of treatment, lipid accumulation, total ROS, mitochondrial ROS, expression and localization of NOX4, and expressions of inflammasomes and IL-1β were detected in the hepatocytes.Compared with the control cells, PA treatment of the cells significantly increased cytoplasmic lipid accumulation, concentrations of total ROS (12 463.09±2.72 vs 6691.23±2.45, P=0.00) and mitochondrial ROS (64.98±0.94 vs 45.04±0.92, P=0.00), and the expressions of NOX4, NLRP3, ASC, caspase-1, and IL-1β (1603.52±1.32 vs 2629.33±2.57, P=0.00). The and NOX4 were found to be co-localized in the cytoplasm. NAC obviously reduced cellular ROS level stimulated by PA (7782.15±2.87 vs 5445.6±1.17, P=0.00) and suppressed the expressions of NLRP3, ASC and caspase-1.PA treatment can stimulate lipid accumulation in hepatocytes and induce oxidative stress through NOX4 and pathway to activate inflammasomes and stimulate the secretion of IL-1β.

Keyword: mitochondria

Tibolone Reduces Oxidative Damage and Inflammation in Microglia Stimulated with through Mechanisms Involving Estrogen Receptor Beta.

High concentrations of in plasma increase both the inflammation associated with obesity and the susceptibility to develop a neurodegenerative event. In the brain, the inflammatory response is mediated by activated microglial cells, which undergo morphological and biochemical changes and can directly affect cell viability. Recent evidence shows that the use of estrogenic compounds can control microglia-induced inflammation with promising results. In this study, we explored the actions of the synthetic steroid tibolone on BV-2 microglia cells stimulated with . Our results demonstrated that tibolone increased cell viability and reduced nuclear fragmentation and the production of reactive oxygen species, as well as preserved mitochondrial membrane potential. These effects were accompanied by reduced nuclear translocation of NF-κB p65, upregulation of neuroglobin, and improved antioxidant defense. Furthermore, estrogen receptor beta (ERβ) inhibition partially dampened tibolone\'s protective actions in BV-2 cells stimulated with . In conclusion, tibolone protects BV-2 cells by a mechanism involving ERβ and upregulation of neuroglobin.

Keyword: mitochondria

Trimetazidine prevents palmitate-induced mitochondrial fission and dysfunction in cultured cardiomyocytes.

Metabolic and cardiovascular disease patients have increased plasma levels of lipids and, specifically, of palmitate, which can be toxic for several tissues. Trimetazidine (TMZ), a partial inhibitor of lipid oxidation, has been proposed as a metabolic modulator for several cardiovascular pathologies. However, its mechanism of action is controversial. Given the fact that TMZ is able to alter mitochondrial metabolism, we evaluated the protective role of TMZ on mitochondrial morphology and function in an in vitro model of lipotoxicity induced by palmitate. We treated cultured rat cardiomyocytes with BSA-conjugated palmitate (25 nM free), TMZ (0.1-100 μM), or a combination of both. We evaluated mitochondrial morphology and lipid accumulation by confocal fluorescence microscopy, parameters of mitochondrial metabolism (mitochondrial membrane potential, oxygen consumption rate [OCR], and ATP levels), and ceramide production by mass spectrometry and indirect immunofluorescence. Palmitate promoted mitochondrial fission evidenced by a decrease in mitochondrial volume (50%) and an increase in the number of per cell (80%), whereas TMZ increased mitochondrial volume (39%), and decreased mitochondrial number (56%), suggesting mitochondrial fusion. Palmitate also decreased mitochondrial metabolism (ATP levels and OCR), while TMZ potentiated all the metabolic parameters assessed. Moreover, pretreatment with TMZ protected the cardiomyocytes from palmitate-induced mitochondrial fission and dysfunction. TMZ also increased lipid accumulation in cardiomyocytes, and prevented palmitate-induced ceramide production. Our data show that TMZ protects cardiomyocytes by changing intracellular lipid management. Thus, the beneficial effects of TMZ on patients with different cardiovascular pathologies can be related to modulation of the mitochondrial morphology and function.Copyright © 2014 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Synthesis of docosahexaenoic from eicosapentaenoic in retina neurons protects photoreceptors from oxidative stress.

Oxidative stress is involved in activating photoreceptor death in several retinal degenerations. Docosahexaenoic (DHA), the major polyunsaturated fatty in the retina, protects cultured retina photoreceptors from apoptosis induced by oxidative stress and promotes photoreceptor differentiation. Here, we investigated whether eicosapentaenoic (EPA), a metabolic precursor to DHA, had similar effects and whether retinal neurons could metabolize EPA to DHA. Adding EPA to rat retina neuronal cultures increased opsin expression and protected photoreceptors from apoptosis induced by the oxidants paraquat and hydrogen peroxide (H2 O2 ). , oleic, and arachidonic acids had no protective effect, showing the specificity for DHA. We found that EPA supplementation significantly increased DHA percentage in retinal neurons, but not EPA percentage. Photoreceptors and glial cells expressed Δ6 desaturase (FADS2), which introduces the last double bond in DHA biosynthetic pathway. Pre-treatment of neuronal cultures with CP-24879 hydrochloride, a Δ5/Δ6 desaturase inhibitor, prevented EPA-induced increase in DHA percentage and completely blocked EPA protection and its effect on photoreceptor differentiation. These results suggest that EPA promoted photoreceptor differentiation and rescued photoreceptors from oxidative stress-induced apoptosis through its elongation and desaturation to DHA. Our data show, for the first time, that isolated retinal neurons can synthesize DHA in culture. Docosahexaenoic (DHA), the major polyunsaturated fatty in retina photoreceptors, and its precursor, eicosapentaenoic (EPA) have multiple beneficial effects. Here, we show that retina neurons in vitro express the desaturase FADS2 and can synthesize DHA from EPA. Moreover, addition of EPA to these cultures protects photoreceptors from oxidative stress and promotes their differentiation through its metabolization to DHA.© 2015 International Society for Neurochemistry.

Keyword: mitochondria

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: mitochondria

NAFLD causes selective CD4(+) T lymphocyte loss and promotes hepatocarcinogenesis.

Hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death. Non-alcoholic fatty liver disease (NAFLD) affects a large proportion of the US population and is considered to be a metabolic predisposition to liver cancer. However, the role of adaptive immune responses in NAFLD-promoted HCC is largely unknown. Here we show, in mouse models and human samples, that dysregulation of lipid metabolism in NAFLD causes a selective loss of intrahepatic CD4(+) but not CD8(+) T lymphocytes, leading to accelerated hepatocarcinogenesis. We also demonstrate that CD4(+) T lymphocytes have greater mitochondrial mass than CD8(+) T lymphocytes and generate higher levels of mitochondrially derived reactive oxygen species (ROS). Disruption of mitochondrial function by linoleic , a fatty accumulated in NAFLD, causes more oxidative damage than other free fatty acids such as , and mediates selective loss of intrahepatic CD4(+) T lymphocytes. In vivo blockade of ROS reversed NAFLD-induced hepatic CD4(+) T lymphocyte decrease and delayed NAFLD-promoted HCC. Our results provide an unexpected link between lipid dysregulation and impaired anti-tumour surveillance.

Keyword: mitochondria

A microplate assay for measuring cell death in C2C12 cells.

The main goal of this study was to develop a straightforward and rapid microplate assay for measuring propidium iodide (PI) in C2C12 cells. The PI method has proven to be an efficient quantitative assay for analyzing cell viability through PI fluorescence analysis. Importantly, the protocol takes less than 30 min and the results are reproducible. C2C12 cells were exposed to an increasing concentration of palmitate for a period of 24 h to induce cell death, and the PI fluorescence increased in a concentration-dependent manner. Evaluation of mitochondrial function and the production of reactive oxygen species confirmed the deleterious effects of palmitate. Also, the microplate PI assay demonstrated high sensitivity, as indicated by the detection of modest fluctuations in cell viability in response to catalase overexpression in palmitate-treated cells. The microplate PI assay, therefore, offers an accurate method for use in in-vitro studies.

Keyword: mitochondria

PINK1-Parkin alleviates metabolic stress induced by obesity in adipose tissue and in 3T3-L1 preadipocytes.

play an important role in cellular metabolism and are closely related with metabolic stress. Recently, several studies have shown that mitophagy mediated by PTEN-induced putative kinase 1 (PINK1) and Parkin may play a critical role in clearing the damaged and maintaining the overall balance of intracellular in quality and quantity. A previous study showed that PINK1 and Parkin were overexpressed in adipose tissue in obese subjects. However, it is still unclear whether a direct relationship exists between obesity and mitophagy. In this study, we created a high-fat-diet (HFD)-induced obese mouse model and examined the expression of PINK1 and Parkin in adipose tissue using western blot and real-time quantitative PCR. After we confirmed that there is an interesting difference between regular-chow-fed mice and HFD-induced obese mice in the expression of PINK1 and Parkin in\xa0vivo, we further tested the expression of PINK1 and Parkin in 3T3-L1 preadipocytes in\xa0vitro by treating cells with (PA) to induce metabolic stress. To better understand the role of PINK1 and Parkin in metabolic stress, 3T3-L1 preadipocytes were transfected with small interfering RNA (siRNA) of PINK1 and Parkin followed by PA treatment. Our results showed that under lower concentrations of PA, PINK1 and Parkin can be activated and play a protective role in resisting the harmful effects of PA, including protecting the mitochondrial function and resisting cellular death, while under higher concentrations of PA, the expression of PINK1 and Parkin can be inhibited. These results suggest that PINK1-Parkin can protect mitochondrial function against metabolic stress induced by obesity or PA to a certain degree.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Proteomic profile of carbonylated proteins in rat liver: discovering possible mechanisms for tetracycline-induced steatosis.

To investigate biochemical mechanisms for the tetracycline-induced steatosis in rats, targeted proteins of oxidative modification were profiled. The results showed that tetracycline induced lipid accumulation, oxidative stress, and cell viability decline in HepG2 cells only under the circumstances of overload. Tetracycline administration in rats led to significant decrement in blood lipids, while resulted in more than four times increment in intrahepatic triacylglycerol and typical microvesicular steatosis in the livers. The triacylglycerol levels were positively correlated with oxidative stress. Proteomic profiles of carbonylated proteins revealed 26 targeted proteins susceptible to oxidative modification and most of them located in . Among them, the long-chain specific acyl-CoA dehydrogenase was one of the key enzymes regulating fatty β-oxidation. Oxidative modification of the enzyme in the tetracycline group depressed its enzymatic activity. In conclusion, the increased influx of lipid into the livers is the first hit of tetracycline-induced microvesicular steatosis. Oxidative stress is an essential part of the second hit, which may arise from the lipid overload and attack a series of functional proteins, aggravating the development of steatosis. The 26 targeted proteins revealed here provide a potential direct link between oxidative stress and tetracycline-induced steatosis.© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: mitochondria

Identification of uncoupling protein 4 from the blood-sucking insect Rhodnius prolixus and its possible role on protection against oxidative stress.

Uncoupling proteins (UCPs) play a critical role in the control of the mitochondrial membrane potential (ΔΨm) due to their ability to dissipate the proton gradient, which results in the uncoupling of mitochondrial respiration from ATP production. Most reactive oxygen species generation in occurs in complex III, due to an increase of semiquinone (Q(-)) half-life. When active, UCPs can account as a potential antioxidant system by decreasing ΔΨm and increasing mitochondrial respiration, thus reducing Q(-) life time. The hematophagous insect Rhodnius prolixus, a vector of Chagas disease, is exposed to a huge increase in oxidative stress after a blood meal because of the hydrolysis of hemoglobin and the release of the cytotoxic heme molecule. Although some protective mechanisms were already described for this insect and other hematophagous arthropods, the putative role of UCP proteins as antioxidants in this context has not been explored. In this report, two genes encoding UCP proteins (RpUcp4 and RpUcp5) were identified in the R.\xa0prolixus genome. RpUcp4 is the predominant transcript in most analyzed organs, and both mRNA and protein expression are upregulated (13- and 3-fold increase, respectively) in enterocytes the first day after the blood feeding. The increase in UCP4 expression is coincident with the decrease in hydrogen peroxide (H2O2) generation by midgut cells. Furthermore, in isolated from enterocytes, the modulation of UCP activity by and GDP resulted in altered ΔΨm, as well as modulation of H2O2 generation rates. These results indicate that R.\xa0prolixus UCP4 may function in an antioxidation mechanism to protect the midgut cells against oxidative damage caused by blood digestion.Copyright © 2014 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Ilexgenin A inhibits endoplasmic reticulum stress and ameliorates endothelial dysfunction via suppression of TXNIP/NLRP3 inflammasome activation in an AMPK dependent manner.

Ilexgenin A is a natural triterpenoid with beneficial effects on lipid disorders. This study aimed to investigate the effects of ilexgenin A on endothelial homeostasis and its mechanisms. Palmitate (PA) stimulation induced endoplasmic reticulum stress (ER stress) and subsequent thioredoxin-interacting protein (TXNIP)/NLRP3 inflammasome activation in endothelial cells, leading to endothelial dysfunction. Ilexgenin A enhanced LKB1-dependent AMPK activity and improved ER stress by suppression of ROS-associated TXNIP induction. However, these effects were blocked by knockdown of AMPKα, indicating AMPK is essential for its action in suppression of ER stress. Meanwhile, ilexgenin A inhibited NLRP3 inflammasome activation by down-regulation of NLRP3 and cleaved caspase-1 induction, and thereby reduced IL-1β secretion. It also inhibited inflammation and apoptosis exposed to PA insult. Consistent with these results in endothelial cells, ilexgenin A attenuated ER stress and restored the loss of eNOS activity in vascular endothelium, and thereby improved endothelium-dependent vasodilation in rat aorta. A further analysis in high-fat fed mice showed that oral administration of ilexgenin A blocked ER stress/NLRP3 activation with reduced ROS generation and increased NO production in vascular endothelium, well confirming the beneficial effect of ilexgenin A on endothelial homeostasis in vivo. Taken together, these results show ER stress-associated TXNIP/NLRP3 inflammasome activation was responsible for endothelial dysfunction and ilexgenin A ameliorated endothelial dysfunction by suppressing ER-stress and TXNIP/NLRP3 inflammasome activation with a regulation of AMPK. This finding suggests that the application of ilexgenin A is useful in the management of cardiovascular diseases in obesity.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Apigenin Ameliorates the Obesity-Induced Skeletal Muscle Atrophy by Attenuating Mitochondrial Dysfunction in the Muscle of Obese Mice.

It was investigated whether apigenin (AP) protected against skeletal muscle atrophy induced by obesity.Mice were fed a high-fat diet (HFD) for 9 weeks to induce obesity, and then were assigned to two groups; the HFD group received a high-fat diet, and the HFD+AP group received a 0.1% AP-containing HFD. After additional feeding of the experimental diet for 8 weeks, mice in the HFD group were highly obese compared with the mice in the standard diet fed mice group. The mice in the AP-treated group showed less fat pad accumulation and less inflammatory cytokines without body weight reduction. The weight of skeletal muscle in the AP group tended to increase as compared with that of the HFD group. Furthermore, AP reduced the expression of atrophic genes, including MuRF1 and Atrogin-1, but increased the exercise capacity. The mitochondrial function and mitochondrial biogenesis were enhanced by AP. In cultured C2C12 cells, AP also suppressed -induced muscle atrophy and mitochondrial dysfunction. In addition, AP activated AMP-activated protein kinase (AMPK) in the C2C12 and the muscle of HFD-induced obese mice.The results suggested that AP ameliorated the obesity-induced skeletal muscle atrophy by attenuating mitochondrial dysfunction.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: mitochondria

Resistance exercise training and in\xa0vitro skeletal muscle oxidative capacity in older adults.

Whether resistance exercise training (RET) improves skeletal muscle substrate oxidative capacity and reduces mitochondrial production of reactive oxygen species in older adults remains unclear. To address this, 19 older males (≥60\xa0years) were randomized to a RET (n\xa0=\xa011) or to a waitlist control group (n\xa0=\xa08) that remained sedentary for 12\xa0weeks. RET was comprised of three upper body and four lower body movements on resistance machines. One set of 8-12 repetitions to failure of each movement was performed on three nonconsecutive days/week. Improvements in chest press and leg press strength were assessed using a three-repetition maximum (3 RM). Body composition was assessed via dual energy X-ray absorptiometry. Muscle biopsies were obtained from the vastus lateralis muscle at baseline and at both 3\xa0weeks and 12\xa0weeks. Palmitate and pyruvate oxidation rates were measured from the (14)CO2 produced from [1-(14)C] and [U-(14)C] pyruvate, respectively, during incubation of muscle homogenates. PGC-1α, TFAM, and PPARδ levels were quantified using qRT-PCR Citrate synthase (CS) and β-HAD activities were determined spectrophotometrically. Mitochondrial production of reactive oxygen species (ROS) were assessed using the Amplex Red Hydrogen Peroxide/Peroxidase assay. There were no significant changes in body weight or body composition following the intervention. Chest press and leg press strength (3RM) increased ~34% (both P\xa0<\xa00.01) with RET There were no significant changes in pyruvate or fatty oxidation or in the expression of target genes with the intervention. There was a modest increase (P\xa0<\xa00.05) in βHAD activity with RET at 12\xa0weeks but the change in CS enzyme activity was not significant. In addition, there were no significant changes in ROS production in either group following RET Taken together, the findings of this study suggest that 12\xa0weeks of low volume RET does not increase skeletal muscle oxidative capacity or reduce ROS production in older adults.© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Keyword: mitochondria

STING-IRF3 Triggers Endothelial Inflammation in Response to Free Fatty -Induced Mitochondrial Damage in Diet-Induced Obesity.

Metabolic stress in obesity induces endothelial inflammation and activation, which initiates adipose tissue inflammation, insulin resistance, and cardiovascular diseases. However, the mechanisms underlying endothelial inflammation induction are not completely understood. Stimulator of interferon genes (STING) is an important molecule in immunity and inflammation. In the present study, we sought to determine the role of STING in -induced endothelial activation/inflammation.In cultured endothelial cells, treatment activated STING, as indicated by its perinuclear translocation and binding to interferon regulatory factor 3 (IRF3), leading to IRF3 phosphorylation and nuclear translocation. The activated IRF3 bound to the promoter of ICAM-1 (intercellular adhesion molecule 1) and induced ICAM-1 expression and monocyte-endothelial cell adhesion. When analyzing the upstream signaling, we found that activated STING by inducing mitochondrial damage. treatment caused mitochondrial damage and leakage of mitochondrial DNA into the cytosol. Through the cytosolic DNA sensor cGAS (cyclic GMP-AMP synthase), the mitochondrial damage and leaked cytosolic mitochondrial DNA activated the STING-IRF3 pathway and increased ICAM-1 expression. In mice with diet-induced obesity, the STING-IRF3 pathway was activated in adipose tissue. However, STING deficiency ( ) partially prevented diet-induced adipose tissue inflammation, obesity, insulin resistance, and glucose intolerance.The mitochondrial damage-cGAS-STING-IRF3 pathway is critically involved in metabolic stress-induced endothelial inflammation. STING may be a potential therapeutic target for preventing cardiovascular diseases and insulin resistance in obese individuals.© 2017 American Heart Association, Inc.

Keyword: mitochondria

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high--diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: mitochondria

PINK1-Parkin-Mediated Mitophagy Protects Mitochondrial Integrity and Prevents Metabolic Stress-Induced Endothelial Injury.

Mitochondrial injury and dysfunction, a significant feature in metabolic syndrome, triggers endothelial cell dysfunction and cell death. Increasing evidence suggests that mitophagy, a process of autophagic turnover of damaged , maintains mitochondrial integrity. PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase 1) and Parkin signaling is a key pathway in mitophagy control. In this study, we examined whether this pathway could protect under metabolic stress. We found that (PA) induced significant mitophagy and activated PINK1 and Parkin in endothelial cells. Knocking down PINK1 or Parkin reduced mitophagy, leading to impaired clearance of damaged and intracellular accumulation of mitochondrial fragments. Furthermore, PINK1 and Parkin prevented PA-induced mitochondrial dysfunction, ROS production and apoptosis. Finally, we show that PINK1 and Parkin were up-regulated in vascular wall of obese mice and diabetic mice. Our study demonstrates that PINK1-Parkin pathway is activated in response to metabolic stress. Through induction of mitophagy, this pathway protects mitochondrial integrity and prevents metabolic stress-induced endothelial injury.

Keyword: mitochondria

Mitochondrial dysfunction and inhibition of myoblast differentiation in mice with high--diet-induced pre-diabetes.

Pre-diabetes is characterized by impaired glucose tolerance (IGT) and/or impaired fasting glucose. Impairment of skeletal muscle function is closely associated with the progression of diabetes. However, the entire pathological characteristics and mechanisms of pre-diabetes in skeletal muscle remain fully unknown. Here, we established a mouse model of pre-diabetes, in which 6-week-old male C57BL6/J mice were fed either normal diet or high- diet (HFD) for 8 or 16 weeks. Both non-fasting and fasting glucose levels and the results of glucose and insulin tolerance tests showed that mice fed an 8-week HFD developed pre-diabetes with IGT; whereas mice fed a 16-week HFD presented with impaired fasting glucose and impaired glucose tolerance (IFG-IGT). Mice at both stages of pre-diabetes displayed decreased numbers of mitochondria in skeletal muscle. Moreover, IFG-IGT mice exhibited decreased mitochondrial membrane potential and ATP production in skeletal muscle and muscle degeneration characterized by a shift in muscle fibers from predominantly oxidative type I to glycolytic type II. Western blotting and histological analysis confirmed that myoblast differentiation was only inhibited in IFG-IGT mice. For primary skeletal muscle satellite cells, inhibition of differentiation was observed in -induced insulin resistance model. Moreover, enhanced myoblast differentiation increased glucose uptake and insulin sensitivity. These findings indicate that pre-diabetes result in mitochondrial dysfunction and inhibition of myoblast differentiation in skeletal muscle. Therefore, interventions that enhance myoblast differentiation may improve insulin resistance of diabetes at the earlier stage.© 2018 Wiley Periodicals, Inc.

Keyword: mitochondria

Short-term cigarette smoke exposure leads to metabolic alterations in lung alveolar cells.

Cigarette smoke (CS)-induced alveolar destruction and energy metabolism changes are known contributors to the pathophysiology of chronic obstructive pulmonary disease (COPD). This study examines the effect of CS exposure on metabolism in alveolar type II cells. Male A/J mice (8 wk old) were exposed to CS generated from a smoking machine for 4 or 8 weeks, and a recovery group was exposed to CS for 8 weeks and allowed to recover for 2 weeks. Alveolar type II cells were isolated from air- or CS- exposed mice. Acute CS exposure led to a reversible airspace enlargement in A/J mice as measured by the increase in mean linear intercept, indicative of alveolar destruction. The effect of CS exposure on cellular respiration was studied using the XF Extracellular Flux Analyzer. A decrease in respiration while metabolizing glucose was observed in the CS-exposed group, indicating altered glycolysis that was compensated by an increase in palmitate utilization; palmitate utilization was accompanied by an increase in the expression of CD36 and carnitine-palmitoyl transferase 1 in type II alveolar cells for the transport of palmitate into the cells and into , respectively. The increase in palmitate use for energy production likely affects the surfactant biosynthesis pathway, as evidenced by the decrease in phosphatidylcholine levels and the increase in phospholipase A2 activity after CS exposure. These findings help our understanding of the mechanism underlying the surfactant deficiency observed in smokers and provide a target to delay the onset of COPD.

Keyword: mitochondria

[THE EXCESS OF FATTY IN FOOD AS MAIN CAUSE OF LIPOIDOSIS OF INSULIN-DEPENDENT CELLS: SKELETAL MYOCYTES, CARDIO-MYOCYTES, PERIPORTAL HEPATOCYTES, KUPFFER MACROPHAGES AND B-CELLS OF PANCREAS].

In phylogenesis, becoming of biologicalfunctions and biological reactions proceeds with the purpose ofpermanent increasing of "kinetic perfection ". The main role belongs to factors ofphysical, chemical and biological kinetics, their evaluation using systemic approach technique under permanent effect of natural selection. The late-in-phylogenesis insulin, proceeded with, in development of biological function of locomotion, specialization of insulin-dependent cells: skeletal myocytes, syncytium of cardiomyocytes, subcutaneous adipocytes, periportal hepatocytes, Kupffer\'s macrophages and β-cells of islets of pancreas. The insulin initiated formation of new, late in phylogenesis, large pool of fatty cells-subcutaneous adipocytes that increased kinetic parameters of biological function of locomotion. In realization of biological function of locomotion only adipocytes absorb exogenous mono unsaturated and saturated fatty acids in the form of triglycerides in composition of oleic and lipoproteins of very low density using apoE/B-100 endocytosis. The rest of insulin-dependent cells absorb fatty acids in the form of unesterified fatty acids from associates with albumin and under effect of CD36 of translocase offatty acids. The insulin in all insulin-depended cells inhibits biological reaction of lipolysis enhancing contributing into development of lipoidosis. The insulin expresses transfer offatty acids in the form of unsaturated fatty acids from adipocytes into matrix of . The insulin supplies insulin-dependent cells with substrates for acquiring energy subject to that in pool of unsaturated fatty acids in adipocytes prevails hydrophobic unsaturated fatiy that slowly passes into matrix through external membrane ofmitochondria; oxidases of so slowly implement its β-oxidation that content of exogenous unsaturatedfatty can\'t be higher than phylogenetic, physiological level - 15% of all amount offatty acids transferring to insulin-dependent cells. The insulin can\'t both to decrease content of exogenous fatty and inhibit lipolysis in visceral fatty cells of omentum.

Keyword: mitochondria

Palmitoleic (16:1n7) increases oxygen consumption, fatty oxidation and ATP content in white adipocytes.

We have recently demonstrated that palmitoleic (16:1n7) increases lipolysis, glucose uptake and glucose utilization for energy production in white adipose cells. In the present study, we tested the hypothesis that palmitoleic modulates bioenergetic activity in white adipocytes.For this, 3\xa0T3-L1 pre-adipocytes were differentiated into mature adipocytes in the presence (or absence) of (16:0) or palmitoleic (16:1n7) at 100 or 200\xa0μM. The following parameters were evaluated: lipolysis, lipogenesis, fatty (FA) oxidation, ATP content, oxygen consumption, mitochondrial mass, citrate synthase activity and protein content of mitochondrial oxidative phosphorylation (OXPHOS) complexes.Treatment with 16:1n7 during 9\xa0days raised basal and isoproterenol-stimulated lipolysis, FA incorporation into triacylglycerol (TAG), FA oxidation, oxygen consumption, protein expression of subunits representing OXPHOS complex II, III, and V and intracellular ATP content. These effects were not observed in adipocytes treated with 16:0.Palmitoleic , by concerted action on lipolysis, FA esterification, mitochondrial FA oxidation, oxygen consumption and ATP content, does enhance white adipocyte energy expenditure and may act as local hormone.

Keyword: mitochondria

Short-term starvation is a strategy to unravel the cellular capacity of oxidizing specific exogenous/endogenous substrates in .

Mitochondrial oxidation of nutrients is tightly regulated in response to the cellular environment and changes in energy demands. studies evaluating the mitochondrial capacity of oxidizing different substrates are important for understanding metabolic shifts in physiological adaptations and pathological conditions, but may be influenced by the nutrients present in the culture medium or by the utilization of endogenous stores. One such influence is exemplified by the Crabtree effect (the glucose-mediated inhibition of mitochondrial respiration) as most experiments are performed in glucose-containing media. Here, using high-resolution respirometry, we evaluated the oxidation of endogenous or exogenous substrates by cell lines harboring different metabolic profiles. We found that a 1-h deprivation of the main energetic nutrients is an appropriate strategy to abolish interference of endogenous or undesirable exogenous substrates with the cellular capacity of oxidizing specific substrates, namely glutamine, pyruvate, glucose, or palmitate, in . This approach primed to immediately increase their oxygen consumption after the addition of the exogenous nutrients. All starved cells could oxidize exogenous glutamine, whereas the capacity for oxidizing palmitate was limited to human hepatocarcinoma Huh7 cells and to C2C12 mouse myoblasts that differentiated into myotubes. In the presence of exogenous glucose, starvation decreased the Crabtree effect in Huh7 and C2C12 cells and abrogated it in mouse neuroblastoma N2A cells. Interestingly, the fact that the Crabtree effect was observed only for mitochondrial basal respiration but not for the maximum respiratory capacity suggests it is not caused by a direct effect on the electron transport system.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: mitochondria

Alleviation of -induced endoplasmic reticulum stress by augmenter of liver regeneration through IP3R-controlled Ca release.

The aberrant release of Ca from the endoplasmic reticulum (ER) contributes to the onset of ER stress, which is closely related to the pathogenesis of non-alcoholic fatty liver disease. We previously reported that augmenter of liver regeneration (ALR) alleviates ER stress and protects hepatocytes from lipotoxicity. However, the link between ALR protection and the suppression of ER stress remains unclear. In this study, we investigated whether the protection against liver steatosis afforded by ALR is related to its inhibition of calcium overflow from the ER to the . The treatment of HepG2 cells with (PA) upregulated IP3R expression, triggering ER-luminal Ca release and inducing ER stress. However, in ALR-transfected (ALR-Tx) HepG2 cells, PA-induced cell injury was clearly alleviated compared with that in vector-Tx cells. After exposure to PA, IP3R expression was downregulated and ER stress was effectively inhibited in the ALR-Tx cells, and ER-Ca release and simultaneous mitochondrial Ca uptake were lower than those in vector-Tx cells. The knockdown of ALR expression with shRNA abolished the protective effects afforded by ALR transfection. PA treatment also suppressed the interaction between BCL-2 and IP3R in HepG2 cells, whereas this interaction was massively enhanced in the ALR-Tx cells, effectively reducing the IP3R-mediated ER-Ca release and thus mitochondrial Ca influx. Our results suggest that the inhibition of ER stress by ALR is related to the interruption of the interaction between BCL2 and IP3R, demonstrating a novel mechanism of ER stress resistance in ALR-Tx cells.© 2018 Wiley Periodicals, Inc.

Keyword: mitochondria

Attenuation of -Induced Lipotoxicity by Chlorogenic through Activation of SIRT1 in Hepatocytes.

Saturated free fatty acids (FFAs) induce hepatocyte lipotoxicity, wherein oxidative stress-associated mitochondrial dysfunction is mechanistically involved. Chlorogenic (CGA), a potent antioxidant and anti-inflammatory compound, protects against high-fat-diet-induced oxidative stress and mitochondrial dysfunction in liver. This study investigates whether CGA protects against FFA-induced hepatocyte lipotoxicity via the regulation of mitochondrial fission/fusion and elucidates its underlying mechanisms.AML12 cell, a non-transformed hepatocyte cell line, is treated with palmitate. Here, it is shown that CGA prevents palmitate-induced lipotoxicity by activation of SIRT1 regulated mitochondrial morphology. CGA treatment mitigates oxidative stress and mitochondrial dysfunction, as evidenced by a decrease in reactive oxygen species (ROS) production, and an increase in mitochondrial mass and mitochondrial membrane potential. CGA also significantly decreases Bax expression and thereby reduces -mediated caspase-dependent apoptosis. Mechanistically, CGA attenuates ROS-induced mitochondrial fragmentation by inhibiting dynamin-related protein 1 (Drp1) and enhancing Mfn2 expression. In contrast, the inhibitory effects of CGA on the generation of mitochondrial ROS and Drp1 are blocked by siRNA knockdown of SIRT1.Collectively, these findings show that supplementation with CGA protects hepatocytes from FFA-induced lipotoxicity through activation of SIRT1, which reverses the oxidative stress and dysfunction of mitochondrial biogenesis directly.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: mitochondria

[The and oleic modes of metabolism of fatty acids. The exogenous syndrome of resistance to insulin under disorder of biologic function of nutrition (trophology)].

The hyperglycemia and insulin are two phylogenetically different humoral regulators of metabolism in vivo. The development of hyperglycemia occurred billions years hitherto under implementation of nutrition function. The insulin was formed in the process of development of biologic function of locomotion. The syndrome of resistance to insulin consists in the derangement of humoral regulation of metabolism of fatty acids and glucose at the phylogenetically different levels in vivo both in paracrine cells cenosis and at the level of organism. The exogenous and endogenic syndromes of resistance to insulin are distinguished. The exogenous resistance to insulin is formed under physiologic function of insulin system when hormone effect is prevented by derangement of biologic function of trophology (nutrition)--the formation of such palmitinic mode of metabolism of fatty acids as substrates for oxidation in . The endogenic syndrome of resistance to insulin consists in discrepancy of regulation of biologic functions at the level of organism under realization of locomotion function and at the level of paracrine cells cenosis under realization of biologic function of adaptation, endoecology (support of "cleanness" of intracellular medium) and its biologic reaction of inflammation, homeostasis function. The syndrome of resistance to insulin is energetic issue in vivo. Primarily, insulin regulates metabolism of fatty acids and only secondly metabolic transformations of glucose. In case ofpalmitinic mode of metabolism offatty acids in the enzymes with the same parameters are involved in biologic reactions. The palmitinic triglycerides are not optimal due to aphysiological slow biochemical and physico-chemical reactions.

Keyword: mitochondria

In vitro treatment of HepG2 cells with saturated fatty acids reproduces mitochondrial dysfunction found in nonalcoholic steatohepatitis.

Activity of the oxidative phosphorylation system (OXPHOS) is decreased in humans and mice with nonalcoholic steatohepatitis. Nitro-oxidative stress seems to be involved in its pathogenesis. The aim of this study was to determine whether fatty acids are implicated in the pathogenesis of this mitochondrial defect. In HepG2 cells, we analyzed the effect of saturated ( and stearic acids) and monounsaturated (oleic ) fatty acids on: OXPHOS activity; levels of protein expression of OXPHOS complexes and their subunits; gene expression and half-life of OXPHOS complexes; nitro-oxidative stress; and NADPH oxidase gene expression and activity. We also studied the effects of inhibiting or silencing NADPH oxidase on the -induced nitro-oxidative stress and subsequent OXPHOS inhibition. Exposure of cultured HepG2 cells to saturated fatty acids resulted in a significant decrease in the OXPHOS activity. This effect was prevented in the presence of a mimic of manganese superoxide dismutase. reduced the amount of both fully-assembled OXPHOS complexes and of complex subunits. This reduction was due mainly to an accelerated degradation of these subunits, which was associated with a 3-tyrosine nitration of mitochondrial proteins. Pretreatment of cells with uric , an antiperoxynitrite agent, prevented protein degradation induced by . A reduced gene expression also contributed to decrease mitochondrial DNA (mtDNA)-encoded subunits. Saturated fatty acids induced oxidative stress and caused mtDNA oxidative damage. This effect was prevented by inhibiting NADPH oxidase. These acids activated NADPH oxidase gene expression and increased NADPH oxidase activity. Silencing this oxidase abrogated totally the inhibitory effect of on OXPHOS complex activity. We conclude that saturated fatty acids caused nitro-oxidative stress, reduced OXPHOS complex half-life and activity, and decreased gene expression of mtDNA-encoded subunits. These effects were mediated by activation of NADPH oxidase. That is, these acids reproduced mitochondrial dysfunction found in humans and animals with nonalcoholic steatohepatitis.© 2015. Published by The Company of Biologists Ltd.

Keyword: mitochondria

Effect of non-esterified fatty acids on fatty metabolism-related genes in calf hepatocytes cultured in vitro.

NEFA plays numerous roles in the metabolism of glucose, lipids, and proteins. A number of experimental studies have shown that NEFA may have an important role in fatty metabolism in the liver, especially in dairy cows that experience negative energy balance (NEB) during early lactation.In this study, using fluorescent quantitative RT-PCR, ELISA, and primary hepatocytes cultured in vitro, we examined the effect of NEFA (0, 0.2, 0.4, 0.8, 1.6, and 3.2 mmol/L) on fatty metabolism by monitoring the mRNA and protein expression of the following key enzymes: long chain acyl-CoA synthetase (ACSL), carnitine palmitoyltransferase IA (CPT IA), long chain acyl-CoA dehydrogenase (ACADL), and acetyl-CoA carboxylase (ACC).The mRNA and protein expression levels of ACSL and ACADL markedly increased as the concentration of NEFA in the media was increased. The mRNA and protein expression levels of CPT IA were enhanced significantly when the NEFA concentrations increased from 0 to 1.6 mmol/L and decreased significantly when the NEFA concentrations increased from 1.6 to 3.2 mmol/L. The mRNA and protein expression of ACC decreased gradually with increasing concentrations of NEFA.These findings indicate that increased NEFA significantly promote the activation and β-oxidation of fatty acids, but very high NEFA concentrations may inhibit the translocation of fatty acids into of hepatocytes. This may explain the development of ketosis or liver lipidosis in dairy cows. CPT IA might be the key control enzyme of the fatty oxidation process in hepatocytes.© 2014 S. Karger AG, Basel.

Keyword: mitochondria

Disruption of the -associated ER membrane (MAM) plays a central role in -induced insulin resistance.

The -associated ER membrane (MAM) is a specialized subdomain of ER that physically connects with . Although disruption of inter-organellar crosstalk via the MAM impairs cellular homeostasis, its pathological significance in insulin resistance in type 2 diabetes mellitus remains unclear. Here, we reveal the importance of reduced MAM formation in the induction of fatty -evoked insulin resistance in hepatocytes. (PA) repressed insulin-stimulated Akt phosphorylation in HepG2 cells within 12h. Treatment with an inhibitor of the ER stress response failed to restore PA-mediated suppression of Akt activation. Mitochondrial reactive oxygen species (ROS) production did not increase in PA-treated cells. Even short-term exposure (3h) to PA reduced the calcium flux from ER to , followed by a significant decrease in MAM contact area, suggesting that PA suppressed the functional interaction between ER and . Forced expression of mitofusin-2, a critical component of the MAM, partially restored MAM contact area and ameliorated the PA-elicited suppression of insulin sensitivity with Ser473 phosphorylation of Akt selectively improved. These results suggest that loss of proximity between ER and , but not perturbation of homeostasis in the two organelles individually, plays crucial roles in PA-evoked Akt inactivation in hepatic insulin resistance.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic fatty liver to nonalcoholic steatohepatitis.

Activation of inflammation is an important mechanism in the development of nonalcoholic steatohepatitis (NASH). This study aims to delineate how mitophagy affects NLRP3 inflammasome activation in hepatic lipotoxicity. Mice were fed a high fat/calorie diet (HFCD) for 24 weeks. Primary rat hepatocytes were treated with (PA) for various periods of time. Mitophagy was measured by protein levels of LC3II and P62. NLRP3, caspase-1, interleukin (IL)-18, and IL-1β at mRNA and protein levels were used as indicators of inflammasome activation. Along with steatotic progression in HFCD-fed mice, ratio of LC3II/β-actin was decreased concurrently with increased levels of liver P62, NLRP3, caspase-1, IL-1β, IL-18, and serum IL-1β levels in late-stage NASH. PA treatment resulted in mitochondrial oxidative stress and initiated mitophagy in primary hepatocytes. The addition of cyclosporine A did not change LC3II/Τοmm20 ratios; but P62 levels were increased after an extended duration of PA exposure, indicating a defect in autophagic activity. Along with impaired mitophagy, mRNA and protein levels of NLRP3, caspase-1, IL-18 and IL-1β were upregulated by PA treatment. Pretreatment with MCC950, N-acetyl cysteine or acetyl-L-carnitine reversed inflammasome activation and a pyroptotic cascade. Additionally, mitophagic flux was partially recovered as indicated by increases in LC3II/Tomm20 ratio, parkin, and PINK1 expression, and decreased P62 expression. The findings suggest that impaired mitophagy triggers hepatic NLRP3 inflammasome activation in a murine NASH model and primary hepatocytes. The new insights into inflammasome activation through mitophagy advance our understanding of how fatty acids elicit lipotoxicity through oxidant stress and autophagy in .

Keyword: mitochondria

Chronic administration of saturated fats and fructose differently affect SREBP activity resulting in different modulation of Nrf2 and Nlrp3 inflammasome pathways in mice liver.

The overconsumption of both saturated fats and fructose in the modern society has been related to the development of nonalcoholic fatty liver disease (NAFLD). However, the specific contribution of individual dietary components on the progression of NAFLD to nonalcoholic steatohepatitis (NASH) has been poorly investigated. Therefore, the aim of our study was to investigate the dissimilar effects of these two dietary components on selected proinflammatory and antioxidant pathways in the liver of C57BL/6 mice fed a standard (SD), a 45% saturated fat (HFAT) or a 60% fructose (HFRT) diet for 12 weeks. HFAT diet evoked systemic metabolic alterations and overweight, not observed in HFRT mice. However, HFRT mice had a greater hepatic triglyceride deposition with increased ratio of triacylglycerols containing the compared to HFAT, as assessed by liquid chromatography-mass spectrometry analysis. This effect is due to the higher activation of the SCAP/SREBP1c lipogenic pathway by HFRT feeding. In addition, we found inhibition of Keap1/Nrf2 antioxidant signaling and more robust stimulation of the Nlrp3 inflammasome pathway in the livers of HFRT-fed mice when compared with HFAT-fed mice, which is consistent with the recent finding that palmitate and SREBP1c are implicated in hepatic oxidative stress and inflammation. These effects were associated with increased hepatic inflammation, as confirmed by high expression of markers of leukocyte infiltration in the HFRT group. Thus, we hypothesize an amplifying loop among lipogenesis, palmitate, Nrf2 and Nlrp3 that leads to a higher risk of NAFLD progression to NASH in a high-fructose diet compared to a high-saturated fat intake.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Ursodeoxycholyl Lysophosphatidylethanolamide modifies aberrant lipid profiles in NAFLD.

Hepatic fat accumulation with disturbed lipid homoeostasis is a hallmark of nonalcoholic fatty liver disease (NAFLD). The bile phospholipid conjugate Ursodeoxycholyl lysophosphatidylethanolamide (UDCA-LPE) is a novel anti-inflammatory agent with hepatoprotective effects in murine high-fat-diet (HFD)-induced NAFLD. The aim of this work was to study changes in the hepatic lipidome due to UDCA-LPE.High fat diet mouse model, mass spectometry, RT-PCR.Hepatic lipid extracts of HFD mice were analysed by mass spectrometry. The results determined higher levels of total, saturated, mono- and diunsaturated fatty acids (FA) in HFD mice, which were decreased by UDCA-LPE predominantly by the reducing the most abundant FA species and oleic . Unlike other FA species, levels of long-chain polyunsaturated fatty acids (LCPUFA), which are composed of arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA), were increased in HFD mice upon UDCA-LPE treatment, mainly due to elevated hepatic ARA pools. Analysis of hepatic phospholipids species showed a decrease in total phosphatidylcholine (PC), especially monounsaturated PC (PUFA-PC) levels in HFD mice. Loss of total PC was reversed due to UDCA-LPE by increasing hepatic PUFA-PC pools. Gene expression analysis showed that UDCA-LPE upregulated PPARα, a key transcriptional regulator of fatty oxidation, as well as downstream target genes CPT1α and AOX, which are crucially involved in mitochondrial and peroxisomal fatty oxidation.UDCA-LPE modulates defective fatty metabolism during experimental NAFLD thereby restoring altered lipid profiles in addition to its pronounced anti-inflammatory effects. Thus, UDCA-LPE may be a promising drug candidate for the management of NAFLD.© 2015 Stichting European Society for Clinical Investigation Journal Foundation.

Keyword: mitochondria

Oleate prevents palmitate-induced mitochondrial dysfunction, insulin resistance and inflammatory signaling in neuronal cells.

Elevated circulating levels of saturated free fatty acids (sFFAs; e.g. palmitate) are known to provoke inflammatory responses and cause insulin resistance in peripheral tissue. By contrast, mono- or poly-unsaturated FFAs are protective against sFFAs. An excess of sFFAs in the brain circulation may also trigger neuroinflammation and insulin resistance, however the underlying signaling changes have not been clarified in neuronal cells. In the present study, we examined the effects of palmitate on mitochondrial function and viability as well as on intracellular insulin and nuclear factor-κB (NF-κB) signaling pathways in Neuro-2a and primary rat cortical neurons. We next tested whether oleate preconditioning has a protective effect against palmitate-induced toxicity. Palmitate induced both mitochondrial dysfunction and insulin resistance while promoting the phosphorylation of mitogen-activated protein kinases and nuclear translocation of NF-κB p65. Oleate pre-exposure and then removal was sufficient to completely block subsequent palmitate-induced intracellular signaling and metabolic derangements. Oleate also prevented ceramide-induced insulin resistance. Moreover, oleate stimulated ATP while decreasing mitochondrial superoxide productions. The latter were associated with increased levels of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α). Inhibition of protein kinase A (PKA) attenuated the protective effect of oleate against palmitate, implicating PKA in the mechanism of oleate action. Oleate increased triglyceride and blocked palmitate-induced diacylglycerol accumulations. Oleate preconditioning was superior to docosahexaenoic (DHA) or linoleate in the protection of neuronal cells against palmitate- or ceramide-induced cytotoxicity. We conclude that oleate has beneficial properties against sFFA and ceramide models of insulin resistance-associated damage to neuronal cells.Copyright © 2014 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Furanoid F- F6 Uniquely Induces NETosis Compared to C16 and C18 Fatty Acids in Human Neutrophils.

Various biomolecules induce neutrophil extracellular trap (NET) formation or NETosis. However, the effect of fatty acids on NETosis has not been clearly established. In this study, we focused on the NETosis-inducing ability of several molecules. We extracted the molecules present in Arabian Gulf catfish (, Val) skin gel, which has multiple therapeutic activities. Gas chromatography⁻mass spectrometry (GC-MS) analysis of the fraction-3 from the gel with NETosis-inducing activity contained fatty acids including a furanoid F- (F6; 12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic ) and common long-chain fatty acids such as (PA; C16:0), palmitoleic (PO; C16:1), stearic (SA; C18:0), and oleic (OA; C18:1). Using pure molecules, we show that all of these fatty acids induce NETosis to different degrees in a dose-dependent fashion. Notably, F6 induces a unique form of NETosis that is rapid and induces reactive oxygen species (ROS) production by both NADPH oxidase (NOX) and mitochondria. F6 also induces citrullination of histone. By contrast, the common fatty acids (PA, PO, SA, and OA) only induce NOX-dependent NETosis. The activation of the kinases such as ERK (extracellular signal-regulated kinase) and JNK (c-Jun N-terminal kinase) is important for long-chain fatty -induced NETosis, whereas, in F--induced NETosis, Akt is additionally needed. Nevertheless, NETosis induced by all of these compounds requires the final chromatin decondensation step of transcriptional firing. These findings are useful for understanding F-- and other fatty -induced NETosis and to establish the active ingredients with therapeutic potential for regulating diseases involving NET formation.

Keyword: mitochondria

Genomics of -laden human hepatocyte cultures enables drug target screening for the treatment of non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a major health burden in need for new medication. To identify potential drug targets a genomic study was performed in -laden primary human hepatocyte (PHH) and human hepatoma cell cultures.PHH, HuH7 and HepG2 hepatoma cell cultures were treated with lipids and/or TNFα. Intracellular load was quantified with the ORO assay. The Affymetrix HG-U133+ array system was employed to perform transcriptome analysis. The droplet (LD) growth and fusion was determined by fluorescence microscopy. LD associated proteins were imaged by confocal immunofluorescence microscopy and confirmed by Western immunoblotting. Bioinformatics defined perturbed metabolic pathways.Whole genome expression profiling identified 227, 1031 and 571 significant regulated genes. Likewise, the combined and TNFα treatment of PHH, HuH7 and HepG2 cell cultures revealed 154, 1238 and 278 differentially expressed genes. Although genomic responses differed among in-vitro systems, commonalities were ascertained by filtering the data for LD associated gene regulations. Among others the LD-growth and fusion associated cell death inducing DFFA like effector C (CIDEC), perilipins (PLIN2, PLIN3), the synaptosome-associated-protein 23 and the vesicle associated membrane protein 3 were strongly up-regulated. Likewise, the PPAR targets pyruvate-dehydrogenase-kinase-4 and angiopoietin-like-4 were up-regulated as was hypoxia-inducible droplet-associated (HILPDA), flotilin and FGF21. Their inhibition ameliorates triglyceride and cholesterol accumulation. TNFα treatment elicited strong induction of the chemokine CXCL8, the kinases MAP3K8, MAP4K4 and negative regulators of cytokine signaling, i.e. SOCS2&SOCS3. Live cell imaging of DsRED calreticulin plasmid transfected HuH7 cells permitted an assessment of LD growth and fusion and confocal immunofluorescence microscopy evidenced induced LD-associated PLIN2, CIDEC, HIF1α, HILPDA, JAK1, PDK4 and ROCK2 expression. Notwithstanding, CPT1A protein was repressed to protect mitochondria from overload. Pharmacological inhibition of the GTPase-dynamin and the fatty transporter-2 reduced uptake by 28.5 and 35%, respectively. Finally, a comparisons of in-vitro/NAFLD patient biopsy findings confirmed common gene regulations thus demonstrating clinical relevance.The genomics of -laden hepatocytes revealed LD-associated gene regulations and perturbed metabolic pathways. Immunofluorescence microscopy confirmed expression of coded proteins to provide a rationale for therapeutic intervention strategies. Collectively, the in-vitro system permits testing of drug candidates.

Keyword: mitochondria

Silymarin protects against renal injury through normalization of lipid metabolism and mitochondrial biogenesis in high fat-fed mice.

Obesity is associated with an increased risk of chronic kidney diseases and the conventional treatment with renin-angiotensin-aldosterone system (RAAS) inhibitors is not enough to prevent renal injury and prolong the progression of disease. Recently, silymarin has shown protective effects on renal tissue injury, but the underlying mechanisms remain elusive. The goal of this study was to investigate the potential capacity of silymarin to prevent renal injury during obesity induced by high fat diet (HFD) in mice. In vivo, male C57BL/6 mice received HFD (60% of total calories) for 12 weeks, randomized and treated orally with vehicle saline or silymarin (30mg/kg body weight/d) for 4 weeks. In vitro, human proximal tubular epithelial cells (HK2) were exposed to 300μM (PA) for 36h followed by silymarin administration at different concentrations. The administration of silymarin significantly ameliorated HFD induced glucose metabolic disorders, oxidative stress and pathological alterations in the kidney. Silymarin significantly mitigated renal lipid accumulation, fatty β-oxidation and mitochondrial biogenesis in HFD mice and PA treated HK2 cells. Furthermore, silymarin partly restored mitochondrial membrane potential of HK2 cells after PA exposure. In conclusion, silymarin can improve oxidative stress and preserve mitochondrial dysfunction in the kidney, potentially via preventing accumulation of renal lipids and fatty β-oxidation.Copyright © 2017. Published by Elsevier Inc.

Keyword: mitochondria

Inhibition of NLRP3 inflammasome by thioredoxin-interacting protein in mouse Kupffer cells as a regulatory mechanism for non-alcoholic fatty liver disease development.

NOD-like receptor (NLR) NLRP3 inflammasome activation has been implicated in the progression of non-alcoholic fatty liver disease (NAFLD) from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH). It has been also shown that (PA) activates NLRP3 inflammasome and promotes interleukin-1β (IL-1β) secretion in Kupffer cells (KCs). However, the specific mechanism of the NLRP3 inflammasome activation is unclear. We studies the molecular mechanisms by investigating the roles of Thioredoxin-interacting protein (TXNIP) and NLRP3 on NAFLD development in patients, high-fat diet (HFD)-induced NAFL and methionine choline deficient (MCD) diet-induced NASH in wild type (WT), TXNIP-/- (thioredoxin-interacting protein) and NLRP3-/- mice, and isolated KCs. We found that the expressions of NLRP3 and TXNIP in human liver tissues were higher in NASH group than in NAFL group. Furthermore, co-immunoprecipitation analyses show that activation of the TXNIP-NLRP3 inflammasome protein complex occurred in KCs of NASH WT mice rather than NAFL WT mice, thus suggesting that the formation and activation of this protein complex is mainly involved in the development of NASH. NLRP3-/- mice exhibited less severe NASH than WT mice in MCD diet model, whereas TXNIP deficiency enhanced NLRP3 inflammasome activation and exacerbated liver injury. PA triggered the activation and co-localization of the NLRP3 inflammasome protein complex in KCs isolated from WT and TXNIP-/- but not NLRP3-/- mice, and most of the complex co-localized with of KCs following PA stimulation. Taken together, our novel findings indicate that TXNIP plays a protective and anti-inflammatory role in the development of NAFLD through binding and suppressing NLRP3.

Keyword: mitochondria

Nrf2 affects the efficiency of mitochondrial fatty oxidation.

Transcription factor Nrf2 (NF-E2 p45-related factor 2) regulates the cellular redox homoeostasis and cytoprotective responses, allowing adaptation and survival under conditions of stress. The significance of Nrf2\xa0in intermediary metabolism is also beginning to be recognized. Thus this transcription factor negatively affects fatty synthesis. However, the effect of Nrf2 on fatty oxidation is currently unknown. In the present paper, we report that the mitochondrial oxidation of long-chain () and short-chain (hexanoic) fatty acids is depressed in the absence of Nrf2 and accelerated when Nrf2 is constitutively active. Addition of fatty acids stimulates respiration in heart and liver isolated from wild-type mice. This effect is significantly weaker when Nrf2 is deleted, whereas it is stronger when Nrf2 activity is constitutively high. In the absence of glucose, addition of fatty acids differentially affects the production of ATP in mouse embryonic fibroblasts from wild-type, Nrf2-knockout and Keap1 (Kelch-like ECH-associated protein 1)-knockout mice. In acute tissue slices, the rate of regeneration of FADH2 is reduced when Nrf2 is absent. This metabolic role of Nrf2 on fatty oxidation has implications for chronic disease conditions including cancer, metabolic syndrome and neurodegeneration.

Keyword: mitochondria

Brefeldin A impairs porcine oocyte meiotic maturation via interruption of organelle dynamics.

Brefeldin A (BFA) is a lactone antibiotic synthesized from by several fungi that could block anterograde transport of proteins from endoplasmic reticulum to Golgi apparatus by reversible disruption of the Golgi complex. Previous investigations have shown that BFA induces the apoptosis of cancer cells in mitosis and impairs asymmetric spindle positioning in meiosis. Here, we document that exposure to BFA in porcine oocytes compromises the meiotic maturation via disrupting both nuclear and cytoplasmic maturation. We found that BFA exposure collapsed the cytoskeleton assembly by showing the aberrant spindle organization with misaligned chromosomes and defective actin dynamics. Furthermore, the distribution of both and cortical granules (CGs), two important indexes of cytoplasmic maturation of oocytes, was disturbed following BFA exposure. We finally validated that the localization of ovastacin, a component of CGs that is essential for the postfertilization removal of sperm-binding sites in the zona pellucida, was also perturbed in BFA-exposed oocytes, which might weaken their fertilization capacity. Collectively, these findings indicate that Golgi-mediated protein transport is indispensable for the porcine oocyte meiotic maturation.© 2019 Wiley Periodicals, Inc.

Keyword: mitochondria

Challenging of AS160/TBC1D4 Alters Intracellular Lipid milieu in L6 Myotubes Incubated With Palmitate.

The Akt substrate of 160\u2009kDa (AS160) is a key regulator of GLUT4 translocation from intracellular depots to the plasma membrane in myocytes. Likely, AS160 also controls LCFAs transport, which requires relocation of fatty transporters. The aim of the present study was to determine the impact of AS160 knockdown on lipid milieu in L6 myotubes incubated with palmitate (PA). Therefore, we compared two different settings, namely: 1) AS160 knockdown prior to palmitate incubation (pre-PA-silencing, AS160 /PA); 2) palmitate incubation with subsequent AS160 knockdown (post-PA-silencing, PA/AS160 ). The efficiency of AS160 silencing was checked at mRNA and protein levels. The expression and localization of FA transporters were determined using Western Blot and immunofluorescence analyses. Intracellular lipid content (FFA, DAG, TAG, and PL) and FA composition were estimated by GLC, whereas basal palmitate uptake was analyzed by means of scintigraphy. Both groups with silenced AS160 were characterized by a greater expression of FA transporters (FAT/CD36, FATP-1, 4) which had contributed to an increased FA cellular influx. Accordingly, we observed that post-PA-silencing of AS160 resulted in a marked decrement in DAG, TAG, and PL contents, but increased FFA content (PA/AS160 vs. PA). The opposite effect was observed in the group with pre-PA-silencing of AS160 in which AS160 knockdown did not affect the lipid pools (AS160 /PA vs. PA). Our results indicate that post-PA-silencing of AS160 has a capacity to decrease the lipotoxic effect(s) of PA by decreasing the content of lipids (DAG and PL) that promote insulin resistance in myotubes. J. Cell. Physiol. 232: 2373-2386, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc.© 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc.

Keyword: mitochondria

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial energy supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in lipid metabolism during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin metabolism. We further demonstrated the impact of cellular lipid metabolism, and particularly cardiolipin metabolism, during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: mitochondria

Hepatocyte miR-33a mediates mitochondrial dysfunction and hepatosteatosis by suppressing NDUFA5.

Emerging evidence suggests that microRNAs (miRNAs) are essential for metabolic haemostasis of liver tissues. Among them, miR-33a is supposed to modulate the cholesterol export and fatty oxidation, but whether miR-33a involves in the process of fatty liver disease is unclear. To disclose the hypothesis, we utilized miR-33a mimic and antisense to explore their effects in primary hepatocytes or high- diet (HFD)-fed mice. Treatment with (PA) or HFD significantly increased the expression of miR-33a in hepatocytes or liver tissues. In primary hepatocytes, miR-33a mimic decreased mitochondrial function, including reduction of ATP production and oxygen consumption, whereas miR-33a inhibition protected PA-induced mitochondrial dysfunction. Interestingly, miR-33a selectively suppressed mitochondrial complex I activity and protein expression, but not other complexes. Through bioinformatics prediction, we found miR-33a directly targeted on the 3\'-UTR of NDUFA5. Dual-luciferase reporter analysis further confirmed the direct suppression of miR-33a on NDUFA5 expression. More importantly, administration of miR-33a antisense could effectively restore HFD-induced mitochondrial dysfunction through up-regulation of NDUFA5 levels. Mice treated with miR-33a antisense also exhibited improved liver function and structural disorders under obese status. Taken together, miR-33a was an important mediator of hepatocyte mitochondrial function, and the therapeutic benefits implied miR-33a antisense had the potential clinical application in combating the fatty liver disease.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: mitochondria

AIF loss deregulates hematopoiesis and reveals different adaptive metabolic responses in bone marrow cells and thymocytes.

Mitochondrial metabolism is a tightly regulated process that plays a central role throughout the lifespan of hematopoietic cells. Herein, we analyze the consequences of the mitochondrial oxidative phosphorylation (OXPHOS)/metabolism disorder associated with the cell-specific hematopoietic ablation of apoptosis-inducing factor (AIF). AIF-null (AIF ) mice developed pancytopenia that was associated with hypocellular bone marrow (BM) and thymus atrophy. Although myeloid cells were relatively spared, the B-cell and erythroid lineages were altered with increased frequencies of precursor B cells, pro-erythroblasts I, and basophilic erythroblasts II. T-cell populations were dramatically reduced with a thymopoiesis blockade at a double negative (DN) immature state, with DN1 accumulation and delayed DN2/DN3 and DN3/DN4 transitions. In BM cells, the OXPHOS/metabolism dysfunction provoked by the loss of AIF was counterbalanced by the augmentation of the mitochondrial biogenesis and a shift towards anaerobic glycolysis. Nevertheless, in a caspase-independent process, the resulting excess of reactive oxygen species compromised the viability of the hematopoietic stem cells (HSC) and progenitors. This led to the progressive exhaustion of the HSC pool, a reduced capacity of the BM progenitors to differentiate into colonies in methylcellulose assays, and the absence of cell-autonomous HSC repopulating potential in vivo. In contrast to BM cells, AIF thymocytes compensated for the OXPHOS breakdown by enhancing fatty β-oxidation. By over-expressing CPT1, ACADL and PDK4, three key enzymes facilitating fatty β-oxidation (e.g., assimilation), the AIF thymocytes retrieved the ATP levels of the AIF cells. As a consequence, it was possible to significantly reestablish AIF thymopoiesis in vivo by feeding the animals with a high-fat diet complemented with an antioxidant. Overall, our data reveal that the mitochondrial signals regulated by AIF are critical to hematopoietic decision-making. Emerging as a link between mitochondrial metabolism and hematopoietic cell fate, AIF-mediated OXPHOS regulation represents a target for the development of new immunomodulatory therapeutics.

Keyword: mitochondria

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: mitochondria

Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells.

Acetyl-CoA carboxylases (ACC) 1 and 2 catalyze the carboxylation of acetyl-CoA to malonyl-CoA and depend on biotin as a coenzyme. ACC1 localizes in the cytoplasm and produces malonyl-CoA for fatty (FA) synthesis. ACC2 localizes in the outer mitochondrial membrane and produces malonyl-CoA that inhibits FA import into for subsequent oxidation. We hypothesized that ACCs are checkpoints in adipocyte differentiation and tested this hypothesis using the ACC1 and ACC2 inhibitor soraphen A (SA) in murine 3T3-L1 preadipocytes. When 3T3-L1 cells were treated with 100nM SA for 8 days after induction of differentiation, the expression of PPARγ mRNA and FABP4 mRNA decreased by 40% and 50%, respectively, compared with solvent controls; the decrease in gene expression was accompanied by a decrease in FABP4 protein expression and associated with a decrease in lipid droplet accumulation. The rate of FA oxidation was 300% greater in SA-treated cells compared with vehicle controls. Treatment with exogenous palmitate restored PPARγ and FABP4 mRNA expression and FABP4 protein expression in SA-treated cells. In contrast, SA did not alter lipid accumulation if treatment was initiated on day eight after induction of differentiation. We conclude that loss of ACC1-dependent FA synthesis and loss of ACC2-dependent inhibition of FA oxidation prevent lipid accumulation in adipocytes and inhibit early stages of adipocyte differentiation.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Altered lipid metabolism in residual white adipose tissues of Bscl2 deficient mice.

Mutations in BSCL2 underlie human congenital generalized lipodystrophy type 2 disease. We previously reported that Bscl2 (-/-) mice develop lipodystrophy of white adipose tissue (WAT) due to unbridled lipolysis. The residual epididymal WAT (EWAT) displays a browning phenotype with much smaller lipid droplets (LD) and higher expression of brown adipose tissue marker proteins. Here we used targeted lipidomics and gene expression profiling to analyze lipid profiles as well as genes involved in lipid metabolism in WAT of wild-type and Bscl2(-/-) mice. Analysis of total saponified fatty acids revealed that the residual EWAT of Bscl2(-/-) mice contained a much higher proportion of oleic 18:1n9 concomitant with a lower proportion of 16:0 , as well as increased n3- polyunsaturated fatty acids (PUFA) remodeling. The acyl chains in major species of triacylglyceride (TG) and diacylglyceride (DG) in the residual EWAT of Bscl2(-/-) mice were also enriched with dietary fatty acids. These changes could be reflected by upregulation of several fatty elongases and desaturases. Meanwhile, Bscl2(-/-) adipocytes from EWAT had increased gene expression in lipid uptake and TG synthesis but not de novo lipogenesis. Both and peroxisomal β-oxidation genes were also markedly increased in Bscl2(-/-) adipocytes, highlighting that these machineries were accelerated to shunt the lipolysis liberated fatty acids through uncoupling to dissipate energy. The residual subcutaneous white adipose tissue (ScWAT) was not browning but displays similar changes in lipid metabolism. Overall, our data emphasize that, other than being essential for adipocyte differentiation, Bscl2 is also important in fatty remodeling and energy homeostasis.

Keyword: mitochondria

Palmitoyl-ceramide accumulation with necrotic cell death in A549 cells, followed by a steep increase in sphinganine content.

Ceramides (Cers) have recently been identified as key signaling molecules that mediate biological functions such as cell growth, differentiation, senescence, apoptosis, and autophagy. However, the functions of Cer accumulation in necrotic cell death remain unknown. The aim of this study was to clarify the relationship between Cer accumulation with inhibition of the conversion pathway of Cer and concomitant necrotic cell death. In order to minimize the effect of apoptosis against necrotic cell death, A549 cells having the inhibiting effect of caspase 9 by survivin were used in this study. Consequently, Cer accumulation in A549 cells would likely be associated with a pathway other than the mitochondrial caspase-dependent pathway of apoptosis. Here, we showed that the dual addition of a glucosyl-Cer synthase inhibitor and a ceramidase inhibitor to A549 cell culture induced palmitoyl-Cer accumulation with Cer synthase 5 expression and necrotic cell death with lysosomal rupture together with leakage of cathepsin B/alkalization after 2-3\xa0h, although it is unknown in this study whether the necrotic cell death was caused by the lysosomal rupture. This Cer accumulation was followed by a steep increase in sphinganine base levels via the activation of serine palmitoyltransferase activity brought about by the increase in palmitoyl-coenzyme A concentration as a substrate after 5-6\xa0h. The increase in palmitoyl-coenzyme A concentration was achieved by activation of the fatty synthetic pathway from acetyl coenzyme A.

Keyword: mitochondria

Lipid composition of the canine sperm plasma membrane as markers of sperm motility.

The fatty composition of the sperm membrane is an important factor involved in the overall sperm quality, including motility. However, in the canine species, the exact composition of the plasma membrane is still unknown. Therefore, the purpose of this study was to evaluate the plasma membrane lipid composition of motile sperm cells and to compare it with asthenospermic samples, as an attempt to determine possible involvements of membrane lipids in dog sperm cell motility. The sperm-rich fraction of ten mature dogs was collected, and samples were subjected to density gradient centrifugation by Percoll , in order to separate motile and asthenospermic samples. Processed semen samples were evaluated for sperm motility, plasma and acrosome membrane integrity, mitochondrial activity and susceptibility to oxidative stress. Lipid plasma membrane composition was identified by mass spectrometry (MALDI-MS). The motile sperm samples presented the following phospholipids in a high frequency in the plasma membrane: phosphatidylcholine 38:4 (composed of stearic and arachidonic fatty acids), phosphatidylcholine 36:1 (stearic and oleic fatty acids), phosphatidylethanolamine 34:4 (myristic and arachidonic fatty acids), glycerophosphatidic 36:4 ( and arachidonic fatty acids), phosphatidylcholine 40:4 plasmanyl and phosphatidylcholine 40:5 plasmenyl. Furthermore, no lipid markers were found in the asthenospermic samples. Results also indicate that differences on plasma membrane composition between motile and asthenospermic samples are crucial factors for determining sperm motility, sperm functionality and susceptibility to oxidative stress. In conclusion, plasma membrane lipid composition varies considerable between motile and asthenospermic samples. Therefore, lipid markers of sperm motility can be considered, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine plasmanyl, phosphatidylcholine plasmenyl and phosphatidic .© 2016 Blackwell Verlag GmbH.

Keyword: mitochondria

Fluxomic evidence for impaired contribution of short-chain acyl-CoA dehydrogenase to mitochondrial palmitate β-oxidation in symptomatic patients with ACADS gene susceptibility variants.

Despite ACADS (acyl-CoA dehydrogenase, short-chain) gene susceptibility variants (c.511C>T and c.625G>A) are considered to be non-pathogenic, encoded proteins are known to exhibit altered kinetics. Whether or not, they might affect overall fatty β-oxidation still remains, however, unclear.De novo biosynthesis of acylcarnitines by whole blood samples incubated with deuterated palmitate (16-H,15-H-palmitate) is suitable as a fluxomic exploration to distinguish between normal and disrupted β-oxidation, abnormal profiles and ratios of acylcarnitines with different chain-lengths being indicative of the site for enzymatic blockade. Determinations in 301 control subjects of ratios between deuterated butyrylcarnitine and sum of deuterated C2 to C14 acylcarnitines served here as reference values to state specifically functional SCAD impairment in patients addressed for clinical and/or biological suspicion of a β-oxidation disorder.Functional SCAD impairment was found in 39 patients. The 27 patients accepting subsequent gene studies were all positive for ACADS mutations. Twenty-six of 27 patients were positive for c.625G>A variant. Twenty-three of 27 patients harbored susceptibility variants as sole ACADS alterations (18 homozygous and 3 heterozygous for c.625G>A, 2 compound heterozygous for c.625G>A/c.511C>T).Our present fluxomic assessment of SCAD suggests a link between ACADS susceptibility variants and abnormal β-oxidation consistent with known altered kinetics of these variants.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Ca(2+)-dependent permeabilization of and liposomes by and oleic acids: a comparative study.

In the present work, we examine and compare the effects of saturated () and unsaturated (oleic) fatty acids in relation to their ability to cause the Ca(2+)-dependent membrane permeabilization. The results obtained can be summarized as follows. (1) Oleic (OA) permeabilizes liposomal membranes at much higher concentrations of Ca(2+) than (PA): 1mM versus 100μM respectively. (2) The OA/Ca(2+)-induced permeabilization of liposomes is not accompanied by changes in the phase state of lipid bilayer, in contrast to what is observed with PA and Ca(2+). (3) The addition of Ca(2+) to the PA-containing vesicles does not change their size; in the case of OA, it leads to the appearance of larger and smaller vesicles, with larger vesicles dominating. This can be interpreted as a result of fusion and fission of liposomes. (4) Like PA, OA is able to induce a Ca(2+)-dependent high-amplitude swelling of , yet it requires higher concentrations of Ca(2+) (30 and 100μM for PA and OA respectively). (5) In contrast to PA, OA is unable to cause the Ca(2+)-dependent high-amplitude swelling of mitoplasts, suggesting that the cause of OA/Ca(2+)-induced permeability transition in may be the fusion of the inner and outer mitochondrial membranes. (6) The presence of OA enhances PA/Ca(2+)-induced permeabilization of liposomes and . The paper discusses possible mechanisms of PA/Ca(2+)- and OA/Ca(2+)-induced membrane permeabilization, the probability of these mechanisms to be realized in the cell, and their possible physiological role.Copyright © 2014 Elsevier B.V. All rights reserved.

Keyword: mitochondria

[THE UNESTERIFIED FATTY ACIDS IN BLOOD PLASMA AND INTERCELLULAR MEDIUM: EFFECT OF INSULIN AND ALBUMIN (THE LECTURE)].

The high content of saturated fatty , triglycerides in food, the large amount of lipoproteins of very low density of the same name in blood, obvious insufficient amount of unesterified fatty acids releasing under lipolysis in blood to meet in vivo requirements in biotransforming energy of ATP are the causes of biological malfunction of homeostasis. As a rule, for every cell in vivo everything is always to be enough. The deficiency of synthesis of ATP by reason of non-optimal substratum for acquirement of ATP by is followed by activation also phylogenetically earlier biological function of adaptation, biological reaction of stress. Thus, surplus of unesterified fatty after every food intake forms in vivo biological reaction of "metabolic" stress, deficiency of energy by reason of realization by in vivo non-optimal exogenous substratum- unesterified fatty , deficiency of acyl- and acetyl-KoA and prognostically formation of potentially ineffective alternative of metabolism of fatty acids. The deficiency of unesterified fatty acids in biological reaction of exotrophy after every intake of food compensates biological reaction of stress, activation of releasing of unesterified fatty acids from visceral fatty cells of gland as it physiologically occurs in biological reaction of endotrophy. At that, adrenalin increases lipolysis in visceral fatty cells of gland and physiologically late insulin can\'t inhibit lipolysis in phylogenetically early visceral fatty cells. Increasing of content of unesterifed fatty acids in blood plasma, as it always occurs in vivo, stops absorption of glucose by cells initiating hyperglycemia, hyperinsulinemia, and syndrome of resistance to insulin. The result of such a compensation of biological reaction of exotrophy is biological reaction of endotrophy, condition of "metabolic" stress, depletion of function of β-cells of islets with formation of diabetes mellitus type I, deficiency in vivo of insulin synthesis. The biological role of albumin - transfer of fatty acids in intercellular medium inform of unesterifed fatty acids and prevention of formation of pool of free fatty acids effecting aphysiologically.

Keyword: mitochondria

Dependence of brown adipose tissue function on CD36-mediated coenzyme Q uptake.

Brown adipose tissue (BAT) possesses the inherent ability to dissipate metabolic energy as heat through uncoupled mitochondrial respiration. An essential component of the mitochondrial electron transport chain is coenzyme Q (CoQ). While cells synthesize CoQ mostly endogenously, exogenous supplementation with CoQ has been successful as a therapy for patients with CoQ deficiency. However, which tissues depend on exogenous CoQ uptake as well as the mechanism by which CoQ is taken up by cells and the role of this process in BAT function are not well understood. Here, we report that the scavenger receptor CD36 drives the uptake of CoQ by BAT and is required for normal BAT function. BAT from mice lacking CD36 displays CoQ deficiency, impaired CoQ uptake, hypertrophy, altered lipid metabolism, mitochondrial dysfunction, and defective nonshivering thermogenesis. Together, these data reveal an important new role for the systemic transport of CoQ to BAT and its function in thermogenesis.Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: mitochondria

Aleuritolic Impaired Autophagic Flux and Induced Apoptosis in Hepatocellular Carcinoma HepG2 Cells.

Aleuritolic (AA) is a triterpene that is isolated from the root of Geisel. In the present study, the cytotoxic effects of AA on hepatocellular carcinoma cells were evaluated. AA exerted dose- and time-dependent cytotoxicity by inducing -dependent apoptosis in the hepatocellular carcinoma cell line, HepG2. Meanwhile, treatment with AA also caused dysregulation of autophagy, as evidenced by enhanced conversion of LC3-I to LC3-II, p62 accumulation, and co-localization of GFP and mCherry-tagged LC3 puncta. Notably, blockage of autophagosome formation by ATG5 knockdown or inhibitors of phosphatidylinositol 3-kinase (3-MA or Ly294002), significantly reversed AA-mediated cytotoxicity. These data indicated that AA retarded the clearance of autophagic cargos, resulting in the production of cytotoxic factors and led to apoptosis in hepatocellular carcinoma cells.

Keyword: mitochondria

Lipotoxicity augments glucotoxicity-induced mitochondrial damage in the development of diabetic retinopathy.

Although hyperglycemia is the main instigator in the development of diabetic retinopathy, dyslipidemia is also considered to play an important role. In the pathogenesis of diabetic retinopathy, cytosolic NADPH oxidase 2 (Nox2) is activated before retinal are damaged. Our aim was to investigate the effect of lipids in the development of diabetic retinopathy.Reactive oxygen species (ROS, by 2\',7\'-dichlorofluorescein diacetate) and activities of Nox2 (by a lucigenin-based method) and Rac1 (by G-LISA) were quantified in retinal endothelial cells incubated with 50 μM palmitate in 5 mM glucose (lipotoxicity) or 20 mM glucose (glucolipotoxicity) for 6 to 96 hours. Mitochondrial DNA (mtDNA) damage was evaluated by extended-length PCR and its transcription by quantifying cytochrome b transcripts.Within 6 hours of exposure of endothelial cells to lipotoxicity, or glucotoxicity (20 mM glucose, without palmitate), significant increase in ROS, Nox2, and Rac1 was observed, which was exacerbated by glucolipotoxic insult. At 48 hours, neither lipotoxicity nor glucotoxicity had any effect on mtDNA and its transcription, but glucolipotoxicity significantly damaged mtDNA and decreased cytochrome b transcripts, and at 96 hours, glucotoxicity and glucolipotoxicity produced similar detrimental effects on mitochondrial damage.Although during initial exposure, lipotoxic or glucotoxic insult produces similar increase in ROS, addition of lipotoxicity in a glucotoxic environment further exacerbates ROS production, and also accelerates their damaging effects on mitochondrial homeostasis. Thus, modulation of Nox2 by pharmacological agents in prediabetic patients with dyslipidemia could retard the development of retinopathy before their hyperglycemia is observable.

Keyword: mitochondria

Smad3 deficiency protects mice from obesity-induced podocyte injury that precedes insulin resistance.

Signaling by TGF-β/Smad3 plays a key role in renal fibrosis. As obesity is one of the major risk factors of chronic and end-stage renal disease, we studied the role of Smad3 signaling in the pathogenesis of obesity-related renal disease. After switching to a high fat diet, the onset of Smad3 C-terminal phosphorylation, increase in albuminuria, and the early stages of peripheral and renal insulin resistance occurred at 1 day, and 4 and 8 weeks, respectively, in C57BL/6 mice. The loss of synaptopodin, a functional marker of podocytes, and phosphorylation of the Smad3 linker region (T179 and S213) appeared after 4 weeks of the high fat diet. This suggests a temporal pattern of Smad3 signaling activation leading to kidney injury and subsequent insulin resistance in the development of obesity-related renal disease. In vivo, Smad3 knockout attenuated the high fat diet-induced proteinuria, renal fibrosis, overall podocyte injury, and mitochondrial dysfunction in podocytes. In vitro palmitate caused a rapid activation of Smad3 in 30\u2009min, loss of synaptopodin in 2 days, and impaired insulin signaling in 3 days in isolated mouse podocytes. Blockade of either Smad3 phosphorylation by SIS3 (a Smad3 inhibitor) or T179 phosphorylation by flavopiridol (a CDK9 inhibitor) prevented the palmitate-induced loss of synaptopodin and mitochondrial function in podocytes. Thus, Smad3 signaling plays essential roles in obesity-related renal disease and may be a novel therapeutic target.

Keyword: mitochondria

Insulin acutely improves mitochondrial function of rat and human skeletal muscle by increasing coupling efficiency of oxidative phosphorylation.

Insulin is essential for the regulation of fuel metabolism and triggers the uptake of glucose by skeletal muscle. The imported glucose is either stored or broken down, as insulin stimulates glycogenesis and ATP synthesis. The mechanism by which ATP production is increased is incompletely understood at present and, generally, relatively little functional information is available on the effect of insulin on mitochondrial function. In this paper we have exploited extracellular flux technology to investigate insulin effects on the bioenergetics of rat (L6) and human skeletal muscle myoblasts and myotubes. We demonstrate that a 20-min insulin exposure significantly increases (i) the cell respiratory control ratio, (ii) the coupling efficiency of oxidative phosphorylation, and (iii) the glucose sensitivity of anaerobic glycolysis. The improvement of mitochondrial function is explained by an insulin-induced immediate decrease of mitochondrial proton leak. Palmitate exposure annuls the beneficial mitochondrial effects of insulin. Our data improve the mechanistic understanding of insulin-stimulated ATP synthesis, and reveal a hitherto undisclosed insulin sensitivity of cellular bioenergetics that suggests a novel way of detecting insulin responsiveness of cells.© 2013.

Keyword: mitochondria

Palmitate induces mitochondrial superoxide generation and activates AMPK in podocytes.

Studies have shown that high levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 diabetes. (PA) is the predominant circulating saturated FFA, yet its role in the pathogenesis of diabetic nephropathy (DN) is not clear. Recently, one study suggested that mitochondrial superoxide production is related to AMP-activated protein kinase (AMPK) activity in diabetic mice kidneys. To elucidate the link between PA and oxidative stress and AMPK activity in DN, we compared the cultured murine podocytes exposed to PA and oleic (OA). Incubation of cells with 250\u2009μM PA or OA induced a translocation of CD36, a fatty transport protein, with intracellular lipid accumulation. PA, but not OA, induced mitochondrial superoxide and hydrogen peroxide (H O ) generation in podocytes, as shown by enhanced fluorescence of MitoSOX Red and dichlorofluorescein (DCF), respectively. Costimulation of PA-treated cells with the H O scavenger catalase abolished the PA-induced DCF fluorescence. Only PA induced mitochondrial damage as shown by electron microscopy. The AMPK activity was determined by immunoblotting, measuring the ratio of phosphorylated AMPK (p-AMPK) to total AMPK. Only PA significantly increased the p-AMPK levels compared with controls. Addition of catalase to PA-treated cells did not affect the PA-stimulated p-AMPK levels. Collectively, our results indicate that PA induces mitochondrial superoxide and H O generation in cultured podocytes, which may not be directly linked to AMPK activation. Given that, PA seems to play an important role in the pathogenesis of DN through lipotoxicity initiated by mitochondrial superoxide overproduction.© 2017 Wiley Periodicals, Inc.

Keyword: mitochondria

Association of low GLP-1 with oxidative stress is related to cardiac disease and outcome in patients with type 2 diabetes mellitus: a pilot study.

Oxidative stress (OS) contributes to cardiovascular damage in type 2 diabetes mellitus (T2DM). The peptide glucagon-like peptide-1 (GLP-1) inhibits OS and exerts cardiovascular protective actions. Our aim was to investigate whether cardiac remodeling (CR) and cardiovascular events (CVE) are associated with circulating GLP-1 and biomarkers of OS in T2DM patients. We also studied GLP-1 antioxidant effects in a model of cardiomyocyte lipotoxicity. We examined 72 T2DM patients with no coronary or valve heart disease and 14 nondiabetic subjects. A median of 6 years follow-up information was obtained in 60 patients. Circulating GLP-1, dipeptidyl peptidase-4 activity, and biomarkers of OS were quantified. In T2DM patients, circulating GLP-1 decreased and OS biomarkers increased, compared with nondiabetics. Plasma GLP-1 was inversely correlated with serum 3-nitrotyrosine in T2DM patients. Patients showing high circulating 3-nitrotyrosine and low GLP-1 levels exhibited CR and higher risk for CVE, compared to the remaining patients. In palmitate-stimulated HL-1 cardiomyocytes, GLP-1 reduced cytosolic and mitochondrial oxidative stress, increased mitochondrial ATP synthase expression, partially restored mitochondrial membrane permeability and cytochrome c oxidase activity, blunted leakage of creatine to the extracellular medium, and inhibited oxidative damage in total and mitochondrial DNA. These results suggest that T2DM patients with reduced circulating GLP-1 and exacerbated OS may exhibit CR and be at higher risk for CVE. In addition, GLP-1 exerts antioxidant effects in HL-1 palmitate-overloaded cardiomyocytes. It is proposed that therapies aimed to increase GLP-1 may counteract OS, protect from CR, and prevent CVE in patients with T2DM.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Decreasing mitochondrial fission alleviates hepatic steatosis in a murine model of nonalcoholic fatty liver disease.

produce the majority of cellular ATP through oxidative phosphorylation, and their capacity to do so is influenced by many factors. Mitochondrial morphology is recently suggested as an important contributor in controlling mitochondrial bioenergetics. divide and fuse continuously, which is affected by environmental factors, including metabolic alterations. Underscoring its bioenergetic influence, altered mitochondrial morphology is reported in tissues of patients and in animal models of metabolic dysfunction. In this study, we found that mitochondrial fission plays a vital role in the progression of nonalcoholic fatty liver disease (NAFLD). The development of hepatic steatosis, oxidative/nitrative stress, and hepatic tissue damage, induced by a high-fat diet, were alleviated in genetically manipulated mice suppressing mitochondrial fission. The alleviation of steatosis was recapitulated in primary hepatocytes with the inhibition of mitochondrial fission. Mechanistically, our study indicates that fission inhibition enhances proton leak under conditions of free fatty incubation, implicating bioenergetic change through manipulating mitochondrial fission. Taken together, our results suggest a mechanistic role for mitochondrial fission in the etiology of NAFLD. The efficacy of decreasing mitochondrial fission in the suppression of NAFLD suggests that mitochondrial fission represents a novel target for therapeutic treatment of NAFLD.Copyright © 2014 the American Physiological Society.

Keyword: mitochondria

Toxicity generated through inhibition of pyruvate carboxylase and carnitine palmitoyl transferase-1 is similar to high glucose/palmitate-induced glucolipotoxicity in INS-1 beta cells.

This work was initiated to determine whether toxicity generated through inhibition of mitochondrial fuel metabolism is similar to high glucose/palmitate (HG/PA)-induced glucolipotoxicity. Influx of glucose and free fatty acids into the tricarboxylic (TCA) cycle was inhibited by treatment with the pyruvate carboxylase (PC) inhibitor phenylacetic (PAA) and carnitine palmitoyl transferase-1 (CPT-1) inhibitor etomoxir (Eto), or knockdown of PC and CPT-1. Treatment of PAA/Eto or knockdown of PC/CPT-1 induced apoptotic death in INS-1 beta cells. Similar to HG/PA treatment, PAA/Eto increased endoplasmic reticulum stress responses but decreased the Akt signal. JNK inhibitor or chemical chaperone was protective against both PAA/Eto- and HG/PA-induced cell death. All attempts to reduce [Ca²⁺](i), stimulate lipid metabolism, and increase the TCA cycle intermediate pool protected PAA/Eto-induced death as well as HG/PA-induced death. These data suggest that signals induced from impaired mitochondrial fuel metabolism play a critical role in HG/PA-induced glucolipotoxicity.Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Keyword: mitochondria

Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high- diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high- diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the liver, liver/body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high- diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high- diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Bcl-2 maintains the mitochondrial membrane potential, but fails to affect production of reactive oxygen species and endoplasmic reticulum stress, in sodium palmitate-induced β-cell death.

Sodium palmitate causes apoptosis of β-cells, and the anti-apoptotic protein Bcl-2 has been shown to counteract this event. However, the exact mechanisms that underlie palmitate-induced pancreatic β-cell apoptosis and through which pathway Bcl-2 executes the protective effect are still unclear.A stable Bcl-2-overexpressing RINm5F cell clone (BMG) and its negative control (B45) were exposed to palmitate for up to 8 h, and cell viability, mitochondrial membrane potential (Δψm), reactive oxygen species (ROS) generation, endoplasmic reticulum (ER) stress, and NF-κB activation were studied in time course experiments.Palmitate exposure for 8 h resulted in increased cell death rates, and this event was partially counteracted by Bcl-2. Bcl-2 overexpression promoted in parallel also a delayed induction of GADD153/CHOP and a weaker phosphorylation of BimEL in palmitate-exposed cells. At earlier time points (2-4 h) palmitate exposure resulted in increased generation of ROS, a decrease in mitochondrial membrane potential (Δψm), and a modest increase in the phosphorylation of eIF2α and IRE1α. BMG cells produced similar amounts of ROS and displayed the same eIF2α and IRE1α phosphorylation rates as B45 cells. However, the palmitate-induced dissipation of Δψm was partially counteracted by Bcl-2. In addition, basal NF-κB activity was increased in BMG cells.Our results indicate that Bcl-2 counteracts palmitate-induced β-cell death by maintaining mitochondrial membrane integrity and augmenting NF-κB activity, but not by affecting ROS production and ER stress.

Keyword: mitochondria

Induction of Ca2+-dependent cyclosporin A-insensitive nonspecific permeability of the inner membrane of liver and cytochrome c release by α,ω-hexadecanedioic in media of varying ionic strength.

In liver loaded with Ca2+ or Sr(2+), α,ω-hexadecanedioic (HDA) can induce nonspecific permeability of the inner membrane (mitochondrial pore) by the mechanism insensitive to cyclosporin A (CsA). In this work we studied the effect of ionic strength of the incubation medium on the kinetics of the processes that accompany Ca2+-dependent induction of the mitochondrial pore by fatty : organelle swelling, Ca2+ release from the matrix, changes in transmembrane potential (Δψ) and rate of oxygen consumption, and the release of cytochrome c from the intermembrane space. Two basic incubation media were used: sucrose medium and isotonic ionic medium containing KCl without sucrose. We found that 200 μM Ca2+ and 20 μM HDA in the presence of CsA effectively induce high-amplitude swelling of both in the case of sucrose and in the ionic incubation medium. In the presence of CsA, can rapidly absorb Ca2+ and retain it in the matrix for a while without reducing Δψ. Upon incubation in the ionic medium, retain most of the added Ca2+ in the matrix for a short time without reducing the Δψ. In both cases the addition of HDA to the 2 min after the introduction of Ca2+ leads to the rapid release of these ions from the matrix and total drop in Δψ. The mitochondrial swelling induced by Ca2+ and HDA in non-ionic medium is accompanied by almost maximal stimulation of respiration. Under the same conditions, but during incubation of in the ionic medium, it is necessary to add cytochrome c for significant stimulation of respiration. The mitochondrial swelling induced by Ca2+ and HDA leads to the release of cytochrome c in a larger amount in the case of ionic medium than for the sucrose medium. We conclude that high ionic strength of the incubation medium determines the massive release of cytochrome c from and liberates it from the respiratory chain, which leads to blockade of electron transport along the respiratory chain and consequently to disruption of the energy functions of the organelles.

Keyword: mitochondria

[CHANGES IN THE METABOLISM IN THE MYOCARDIUM OF RATS WITH ARTERIAL HYPERTENSION].

In the myocardium of the rats with arterial hypertension marked increase in the amount of unsaturated fatty acids and polyunsaturated fatty acids. Reducing the concentration of and increased levels of arachidonic is considered as one of the factors that lead to the development of energy deficit and oxidative stress. In rats, with hypertension myocardial lactate concentration increases in the cytoplasmic fraction and reducing the amount of ATP. The level in the cytoplasmic and mitochondrial fractions above benchmarks, indicating about the change of antioxidant systems of the body In the cytoplasm and of cardiomyocytes of the rats with arterial hypertension marked decrease in the activity of antioxidant enzymes: NO-synthase, catalase, glutathione reductase, which causes metabolic changes of the myocardium.

Keyword: mitochondria

Dihydromyricetin inhibits NLRP3 inflammasome-dependent pyroptosis by activating the Nrf2 signaling pathway in vascular endothelial cells.

Increasing evidence demonstrates that pyroptosis, pro-inflammatory programmed cell death, is linked to atherosclerosis; however, the underlying mechanisms remain to be elucidated. Dihydromyricetin (DHM), a natural flavonoid, was reported to exert anti-oxidative and anti-inflammatory bioactivities. However, the effect of DHM on atherosclerosis-related pyroptosis has not been studied. In the present study, (PA) treatment led to pyroptosis in human umbilical vein endothelial cells (HUVECs), as evidenced by caspase-1 activation, LDH release, and propidium iodide-positive staining; enhanced the maturation and release of proinflammatory cytokine IL-1β and activation of the NLRP3 inflammasome; and markedly increased intracellular reactive oxygen species (ROS) and mitochondrial ROS (mtROS) levels. Moreover, NLRP3 siRNA transfection or treatment with inhibitors efficiently suppressed PA-induced pyroptosis, and pretreatment with total ROS scavenger or mtROS scavenger attenuated PA-induced NLRP3 inflammasome activation and subsequent pyroptosis. However, DHM pretreatment inhibited PA-induced pyroptotic cell death by increasing cell viability, decreasing LDH and IL-1β release, improving cell membrane integrity, and abolishing caspase-1 cleavage and subsequent IL-1β maturation. We also found that DHM pre-treatment remarkably reduced the levels of intracellular ROS and mtROS and activated the Nrf2 signaling pathway. Moreover, knockdown of Nrf2 by siRNA abrogated the inhibitory effects of DHM on ROS generation and subsequent PA-induced pyroptosis. Together, these results indicate that the Nrf2 signaling pathway plays a role, as least in part, in the DHM-mediated improvement in PA-induced pyroptosis in vascular endothelial cells, which implies the underlying medicinal value of DHM targeting immune/inflammatory-related diseases, such as atherosclerosis.© 2017 BioFactors, 44(2):123-136, 2018.

Keyword: mitochondria

Activation of Mitochondrial Uncoupling Protein 4 and ATP-Sensitive Potassium Channel Cumulatively Decreases Superoxide Production in Insect .

It has been evidenced that mitochondrial uncoupling protein 4 (UCP4) and ATP-regulated potassium channel (mKATP channel) of insect Gromphadorhina coqereliana decrease superoxide anion production. We elucidated whether the two energy-dissipating systems work together on a modulation of superoxide level in cockroach . Our data show that the simultaneous activation of UCP4 by and mKATP channel by pinacidil revealed a cumulative effect on weakening mitochondrial superoxide formation. The inhibition of UCP4 by GTP (and/or ATP) and mKATP channel by ATP elevated superoxide production. These results suggest a functional cooperation of both energy-dissipating systems in protection against oxidative stress in insects.

Keyword: mitochondria

Uncoupling effect of palmitate is exacerbated in skeletal muscle of sea-acclimatized king penguins (Aptenodytes patagonicus).

In king penguin juveniles, the environmental transition from a terrestrial to a marine habitat, occurring at fledging, drastically stimulates lipid catabolism and the remodelling of muscle to sustain extensive swimming activity and thermoregulation in the cold circumpolar oceans. However, the exact nature of these mechanisms remains only partially resolved. Here we investigated, in vitro, the uncoupling effect of increasing doses of fatty acids in pectoralis muscle intermyofibrillar isolated, either from terrestrial never-immersed or experimentally cold water immersed pre-fledging king penguins or from sea-acclimatized fledged penguins. exhibited much greater palmitate-induced uncoupling respiration and higher maximal oxidative capacity after acclimatization to marine life. Such effects were not reproduced experimentally after repeated immersions in cold water, suggesting that the plasticity of mitochondrial characteristics may not be primarily driven by cold exposure per se but by other aspects of sea acclimatization.Copyright © 2017. Published by Elsevier Inc.

Keyword: mitochondria

Effects of high-fat diet and AMP-activated protein kinase modulation on the regulation of whole-body lipid metabolism.

Metabolic flexibility, the capacity to adapt to fuel availability for energy production, is crucial for maintaining whole-body energy homeostasis. An inability to adequately promote FA utilization is associated with lipid accumulation in peripheral tissues and contributes to the development of insulin resistance. In vivo assays to quantify whole-body lipid oxidation in mouse models of insulin resistance are lacking. We describe a method for assessing whole-body FA oxidation in vivo, as well as tissue-specific lipid uptake in conscious mice. The method relies on intravenous administration of [9,10-H(N)] combined with a non-β-oxidizable palmitate analog, [1-C]2-bromopalmitic . Pretreatment with etomoxir, a CPT1 inhibitor that prevents the shuttling of FAs into , markedly reduced the appearance of the β-oxidation product HO in circulation and reduced lipid uptake by oxidative tissues including heart and soleus muscle. Whole-body fatty oxidation was unaltered between chow- or high-fat-fed WT and transgenic mice expressing a mutant form of the AMPK γ3 subunit (AMPKγ3) in skeletal muscle. High-fat feeding increased lipid oxidation in WT and AMPKγ3 transgenic mice. In conclusion, this technique allows for the assessment of the effect of pharmaceutical agents, as well as gene mutations, on whole-body FA oxidation in mice.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: mitochondria

Polysaccharides Prevent -Evoked Apoptosis and Autophagy in Intestinal Porcine Epithelial Cell Line via Restoration of Mitochondrial Function and Regulation of MAPK and AMPK/Akt/mTOR Signaling Pathway.

polysaccharide (GLP) extracted from (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high-fat constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.

Keyword: mitochondria

Phycocyanin-Functionalized Selenium Nanoparticles Reverse -Induced Pancreatic β Cell Apoptosis by Enhancing Cellular Uptake and Blocking Reactive Oxygen Species (ROS)-Mediated Dysfunction.

Accumulation of (PA) in human bodies could cause damage to pancreatic β cells and lead to chronic diseases by generation of reactive oxygen species (ROS). Therefore, it is of great significance to search for nutrition-available agents with antioxidant activity to protect pancreatic islet cells against PA-induced damage. Phycocyanin (PC) and selenium (Se) have been reported to have excellent antioxidant activity. In this study, PC-functionalized selenium nanoparticles (PC-SeNPs) were synthesized to investigate the in vitro protective effects on INS-1E rat insulinoma β cells against PA-induced cell death. A potent protective effect was achieved by regulation of particle size and PC content. Among three PC-SeNPs (165, 235, and 371 nm), PC-SeNPs-235 nm showed the highest cellular uptake and the best protective activities. For cell cycle analysis, PC-SeNPs showed a better protective effect on PA-induced INS-1E cell apoptosis than PC or SeNPs, and PC-SeNPs-235 nm exhibited the best effect. Further mechanistic studies demonstrated that PA induced overproduction of intracellular ROS, fragmentation, activation of caspase-3, -8, and -9, and cleavage of PARP. However, pretreatment of the cells with PC-SeNPs effectively blocked these intracellular events, which suggests that PC-SeNPs could protect INS-1E cells against PA-induced cell apoptosis via attenuating oxidative stress and downstream signaling pathways. This finding provides a great promising nutritional approach for protection against diseases related to islet damage.

Keyword: mitochondria

Lipotubuloids in ovary epidermis of Ornithogalum umbellatum act as metabolons: suggestion of the name \'lipotubuloid metabolon\'.

A metabolon is a temporary, structural-functional complex formed between sequential metabolic enzymes and cellular elements. Cytoplasmic domains called lipotubuloids are present in Ornithogalum umbellatum ovary epidermis. They consist of numerous lipid bodies entwined with microtubules, polysomes, rough endoplasmic reticulum (RER), and actin filaments connected to microtubules through myosin and kinesin. A few , Golgi structures, and microbodies are also observed and also, at later development stages, autolytic vacuoles. Each lipotubuloid is surrounded by a tonoplast as it invaginates into a vacuole. These structures appear in young cells, which grow intensively reaching 30-fold enlargement but do not divide. They also become larger due to an increasing number of lipid bodies formed in the RER by the accumulation of lipids between leaflets of the phospholipid bilayer. When a cell ceases to grow, the lipotubuloids disintegrate into individual structures. Light and electron microscope studies using filming techniques, autoradiography with [(3)H], immunogold labelling with antibodies against DGAT2, phospholipase D1 and lipase, and double immunogold labelling with antibodies against myosin and kinesin, as well as experiments with propyzamide, a microtubule activity inhibitor, have shown that lipotubuloids are functionally and structurally integrated metabolons [here termed lipotubuloid metabolons (LMs)] occurring temporarily in growing cells. They synthesize lipids in lipid bodies in cooperation with microtubules. Some of these lipids are metabolized and used by the cell as nutrients, and others are transformed into cuticle whose formation is mediated by cutinsomes. The latter were discovered in planta using specific anti-cutinsome antibodies visualized by gold labelling. Moreover, LMs are able to rotate autonomously due to the interaction of microtubules, actin filaments, and motor proteins, which influence microtubules by changing their diameter.© The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Keyword: mitochondria

Tibolone attenuates inflammatory response by and preserves mitochondrial membrane potential in astrocytic cells through estrogen receptor beta.

(PA) induces several metabolic and molecular changes in astrocytes, and, it is involved in pathological conditions related to neurodegenerative diseases. Previously, we demonstrated that tibolone, a synthetic steroid with estrogenic, progestogenic and androgenic actions, protects cells from mitochondrial damage and morphological changes induced by PA. Here, we have evaluated which estrogen receptor is involved in protective actions of tibolone and analyzed whether tibolone reverses gene expression changes induced by PA. Tibolone actions on astrocytic cells were mimicked by agonists of estrogen receptor α (ERα) and β (ERβ), but the blockade of both ERs suggested a predominance of ERβ on membrane potential. Expression analysis showed a significant effect of tibolone on genes associated with inflammation such as IL6, IL1B and miR155-3p. It is noteworthy that tibolone attenuated the increased expression of TERT, TERC and DNMT3B genes induced by . Our results suggest that tibolone has anti-inflammatory effects and can modulate pathways associated with DNA methylation and telomeric complex. However, future studies are needed to elucidate the role of epigenetic mechanisms and telomere-associated proteins on tibolone actions.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: mitochondria

Hepatic stimulator substance inhibits calcium overflow through the -associated membrane compartment during nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease is considered a disorder of the endoplasmic reticulum (ER) and . Recent studies have shown that the ER and mitochondrial membranes overlap by 15-20%, a region referred to as the \'-associated ER membrane\' (MAM). Some proteins, including sarco/ER calcium ATPase (SERCA), are located in the MAM and have an important role in Ca signaling and homeostasis between the ER and the . Our previous study showed that hepatic stimulator substance (HSS) inhibits the ER stress induced by reactive oxygen species, thus reducing mitochondrial damage. However, the mechanism underlying the protective effect of HSS on the ER and ER-mitochondrial interaction remains unclear. In this study, we confirmed that the exogenous expression of HSS protected the liver from steatosis in mice with nonalcoholic steatohepatitis. More importantly, the protection provided by HSS allowed SERCA in the MAM compartment to function well, preventing the extensive influx of cytosolic free Ca to the , thus preserving the mitochondrial functions from calcium overload and relieving -induced hepatocyte steatosis. Our results suggest that the protective effect of HSS on SERCA expression is associated with the maintenance of calcium homeostasis within the MAM, thus ameliorating the disordered Ca communication between the ER and .

Keyword: mitochondria

Acetyl-CoA from inflammation-induced fatty acids oxidation promotes hepatic malate-aspartate shuttle activity and glycolysis.

Hepatic metabolic syndrome is associated with inflammation, as inflammation stimulates the reprogramming of nutrient metabolism and hepatic -generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and fatty acids, in the condition of inflammation is still unclear. Here, we used an acute inflammation model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic glycolysis and fatty oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs liver showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain glycolysis. With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived fatty oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by fatty oxidation promotes MAS activity and glycolysis via nonenzymatic acetylation during the inflammatory stress response.

Keyword: mitochondria

Mitochondrial Metabolic Reprogramming by CD36 Signaling Drives Macrophage Inflammatory Responses.

A hallmark of chronic inflammatory disorders is persistence of pro-inflammatory macrophages in diseased tissues. In atherosclerosis this is associated with dyslipidemia and oxidative stress, but mechanisms linking these phenomena to macrophage activation remain incompletely understood. To investigate mechanisms linking dyslipidemia, oxidative stress and macrophage activation through modulation of immunemetabolism, and to explore therapeutic potential targeting specific metabolic pathways. Using a combination of biochemical, immunological, and ex vivo cell metabolic studies, we report that CD36 mediates a mitochondrial metabolic switch from oxidative phosphorylation to superoxide production in response to its ligand, oxLDL. Mitochondrial-specific inhibition of superoxide inhibited oxLDL-induced NF-κB activation and inflammatory cytokine generation. RNAseq, flow cytometry, H-labeled uptake, lipidomic analysis, confocal and EM imaging, and functional energetics revealed that oxLDL upregulated effectors of long-chain fatty (FA) uptake and mitochondrial import, while downregulating FA oxidation and inhibiting ATP5A, an electron transport chain (ETC) component. The combined effect is long-chain FA accumulation, alteration of mitochondrial structure and function, repurposing of the ETC to superoxide production, and NF-κB activation. Apoe null mice challenged with high fat diet showed similar metabolic changes in circulating Ly6C monocytes and peritoneal macrophages, along with increased CD36 expression. Moreover, mitochondrial ROS was positively correlated with CD36 expression in aortic lesional macrophages. These findings reveal that oxLDL/CD36 signaling in macrophages links dys-regulated FA metabolism to oxidative stress from the , which drives chronic inflammation. Thus, targeting to CD36 and its downstream effectors may serve as potential new strategies against chronic inflammatory diseases such as atherosclerosis.

Keyword: mitochondria

Silencing miR-106b improves -induced mitochondrial dysfunction and insulin resistance in skeletal myocytes.

MicroRNA‑106b (miR‑106b) is reported to correlate closely with skeletal muscle insulin resistance. In the current study the effect of miR‑106b on (PA)‑induced mitochondrial dysfunction and insulin resistance was investigated in C2C12 myotubes via the silencing of miR‑106b. MiR‑106b expression was increased under PA treatment, while miR‑106b loss of function improved insulin sensitivity by upregulating its target mitofusin‑2 (Mfn2) in C2C12 myocytes. Furthermore, miR‑106b loss of function partly improved mitochondrial morphological lesions and increased the levels of mitochondial DNA and intracellular adenosine triphosphate that had been impaired by PA exposure in C2C12 myocytes. MiR‑106b loss of function attenuated the levels of intracellular reactive oxygen species (ROS), and upregulated the expression levels of the estrogen‑related receptor (ERR)‑α/peroxisome proliferative activated receptor γ coactivator (PGC)‑1α/Mfn2 axis under PA exposure. In addition, miR‑106b negatively regulated skeletal muscle mitochondrial function and insulin sensitivity under PA‑induced insulin resistance by targeting Mfn2, which may be associated with reduced ROS and upregulation of the ERR‑α/PGC‑1α/Mfn2 axis.

Keyword: mitochondria

Resveratrol attenuates oxidative injury in human umbilical vein endothelial cells through regulating mitochondrial fusion via TyrRS-PARP1 pathway.

Oxidative stress-induced damage in endothelial cells is a crucial initiator of atherosclerosis (AS), which is highly related to excessive reactive oxygen species (ROS) and mitochondrial dynamics. Resveratrol (RSV) exerts beneficial effects against endothelial oxidative injury, while the underlying mechanisms have not been fully elucidated. Thus, we aimed to explore the role of dynamics during the anti-oxidative activities of RSV in (PA)-stimulated human umbilical vein endothelial cells (HUVECs) and to verify whether tyrosyl transfer- RNA synthetase (TyrRS) and poly (ADP-ribose) polymerase 1 (PARP1) are targeted during this process.HUVECs were exposed to 200\u2009μM of PA for 16\u2009h before treated with 10\u2009μM of RSV for 8\u2009h. Cell viability was detected using Cell counting kit-8 (CCK-8) assay. The intracellular ROS level and membrane potential (MMP) were measured using microplate reader and flow cytometry. The malondialdehyde and superoxide dismutase were measured using the microplate reader. The mitochondrial morphology and fusion process was observed under transmission electron microscopy and confocal microscopy. TyrRS and PARP1 were knocked down with the specific small interference RNAs (siRNA), and the protein expressions of TyrRS, PARP1, and mitochondrial fusion proteins (MFN1, MFN2, and OPA1) were measured by western blot.RSV treatment suppressed the PA-induced injuries in HUVECs, including the damage to cell viability, oxidative stress, and loss of MMP. Additionally, RSV improved the protein levels of MFN1, MFN2, and OPA1 as well as inhibited the PA-induced fragmentation of . However, the effects of RSV on oxidative stress and mitochondrial fusion were abolished by the pretreatment of siRNAs of TyrRS and PARP1, indicating that these effects of RSV were dependent on the TyrRS-PARP1 pathway.RSV attenuated endothelial oxidative injury by regulating mitochondrial fusion via TyrRS-PARP1 signaling pathway.

Keyword: mitochondria

Free fatty receptor 1 (FFAR1/GPR40) signaling affects insulin secretion by enhancing mitochondrial respiration during palmitate exposure.

Fatty acids affect insulin secretion via metabolism and FFAR1-mediated signaling. Recent reports indicate that these two pathways act synergistically. Still it remains unclear how they interrelate. Taking into account the key role of in insulin secretion, we attempted to dissect the metabolic and FFAR1-mediated effects of fatty acids on mitochondrial function. One-hour culture of MIN6 cells with palmitate significantly enhanced mitochondrial respiration. Antagonism or silencing of FFAR1 prevented the palmitate-induced rise in respiration. On the other hand, in the absence of extracellular palmitate FFAR1 agonists caused a modest increase in respiration. Using an agonist of the M3 muscarinic acetylcholine receptor and PKC inhibitor we found that in the presence of the fatty mitochondrial respiration is regulated via Gαq protein-coupled receptor signaling. The increase in respiration in palmitate-treated cells was largely due to increased glucose utilization and oxidation. However, glucose utilization was not dependent on FFAR1 signaling. Collectively, these results indicate that mitochondrial respiration in palmitate-treated cells is enhanced via combined action of intracellular metabolism of the fatty and the Gαq-coupled FFAR1 signaling. Long-term palmitate exposure reduced ATP-coupling efficiency of and deteriorated insulin secretion. The presence of the FFAR1 antagonist during culture did not improve ATP-coupling efficiency, however, it resulted in enhanced mitochondrial respiration and improved insulin secretion after culture. Taken together, our study demonstrates that during palmitate exposure, integrated actions of fatty metabolism and fatty -induced FFAR1 signaling on mitochondrial respiration underlie the synergistic action of the two pathways on insulin secretion.Copyright © 2015. Published by Elsevier B.V.

Keyword: mitochondria

Glucose fluctuation increased hepatocyte apoptosis under lipotoxicity and the involvement of mitochondrial permeability transition opening.

Oxidative stress is considered to be an important factor in producing lethal hepatocyte injury associated with nonalcoholic fatty liver disease (NAFLD). Glucose fluctuation, more pronounced in patients with diabetes, has been recognized as an even stronger oxidative stress inducer than the sustained hyperglycemia. Here, we investigated the role of glucose variability in the development of the NAFLD based on hepatocyte apoptosis and possible mechanisms. To achieve this goal we studied C57BL/6J mice that were maintained on a high fat diet (HFD) and injected with glucose (3 g/kg) twice daily to induce intermittent high glucose (IHG). We also studied hepatic L02 cells incubated with (PA) to induce steatosis. The following experimental groups were compared: normal glucose (NG), sustained high glucose (SHG) and IHG with or without PA. We found that, although hepatic enzyme levels and liver lipid deposition were comparable between HFD mice injected with glucose or saline, the glucose injected mice displayed marked hepatocyte apoptosis and inflammation, accompanied by increased lipid peroxide in liver. In vitro, in the presence of PA, IHG increased L02 cell apoptosis and oxidative stress and produced pronounced mitochondrial dysfunction relative to the NG and SHG groups. Furthermore, treatment with the mitochondrial permeability transition (MPT) inhibitor, cyclosporin A (1.5 μmol/l), prevented mitochondrial dysfunction, oxidative stress and hepatocyte apoptosis. Our data suggests that IHG under lipotoxicity might contribute to the development of NAFLD by increasing oxidative stress and hepatocyte apoptosis via MPT and its related mitochondrial dysfunction.© 2015 Society for Endocrinology.

Keyword: mitochondria

2-Chlorohexadecanoic induces ER stress and mitochondrial dysfunction in brain microvascular endothelial cells.

Peripheral leukocytes induce blood-brain barrier (BBB) dysfunction through the release of cytotoxic mediators. These include hypochlorous (HOCl) that is formed via the myeloperoxidase-HO-chloride system of activated phagocytes. HOCl targets the endogenous pool of ether phospholipids (plasmalogens) generating chlorinated inflammatory mediators like e.g. 2-chlorohexadecanal and its conversion product 2-chlorohexadecanoic (2-ClHA). In the cerebrovasculature these compounds inflict damage to brain microvascular endothelial cells (BMVEC) that form the morphological basis of the BBB. To follow subcellular trafficking of 2-ClHA we synthesized a \'clickable\' alkyne derivative (2-ClHyA) that phenocopied the biological activity of the parent compound. Confocal and superresolution structured illumination microscopy revealed accumulation of 2-ClHyA in the endoplasmic reticulum (ER) and of human BMVEC (hCMEC/D3 cell line). 2-ClHA and its alkyne analogue interfered with protein palmitoylation, induced ER-stress markers, reduced the ER ATP content, and activated transcription and secretion of interleukin (IL)-6 as well as IL-8. 2-ClHA disrupted the mitochondrial membrane potential and induced procaspase-3 and PARP cleavage. The protein kinase R-like ER kinase (PERK) inhibitor GSK2606414 suppressed 2-ClHA-mediated activating transcription factor 4 synthesis and IL-6/8 secretion, but showed no effect on endothelial barrier dysfunction and cleavage of procaspase-3. Our data indicate that 2-ClHA induces potent lipotoxic responses in brain endothelial cells and could have implications in inflammation-induced BBB dysfunction.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Effects of Fatty Acids on Intracellular [Ca2+], Mitochondrial Uncoupling and Apoptosis in Rat Pachytene Spermatocytes and Round Spermatids.

The aim of this work was to explore the ability of free arachidonic , and the unsaturated fatty acids oleic and docosahexaenoic to modify calcium homeostasis and mitochondrial function in rat pachytene spermatocytes and round spermatids. In contrast to , unsaturated fatty acids produced significant increases in intracellular calcium concentrations ([Ca2+]i) in both cell types. Increases were fatty specific, dose-dependent and different for each cell type. The arachidonic effects on [Ca2+]i were higher in spermatids than in spermatocytes and persisted when residual extracellular Ca2+ was chelated by EGTA, indicating that the increase in [Ca2+]i originated from release of intracellular calcium stores. At the concentrations required for these increases, unsaturated fatty acids produced no significant changes in the plasma membrane potential of or non-specific permeability in spermatogenic cells. For the case of arachidonic , the [Ca2+]i increases were not caused by its metabolic conversion to eicosanoids or anandamide; thus we attribute this effect to the fatty itself. As estimated with fluorescent probes, unsaturated fatty acids did not affect the intracellular pH but were able to induce a progressive decrease in the mitochondrial membrane potential. The association of this decrease with reduced reactive oxygen species (ROS) production strongly suggests that unsaturated fatty acids induced mitochondrial uncoupling. This effect was stronger in spermatids than in spermatocytes. As a late event, arachidonic induced caspase 3 activation in a dose-dependent manner both in the absence and presence of external Ca2+. The concurrent but differential effects of unsaturated fatty acids on [Ca2+]i and mitochondrial functions are additional manifestations of the metabolic changes that germ cells undergo during their differentiation.

Keyword: mitochondria

Metformin Restores Parkin-Mediated Mitophagy, Suppressed by Cytosolic p53.

Metformin is known to alleviate hepatosteatosis by inducing 5\' adenosine monophosphate (AMP)-kinase-independent, sirtuin 1 (SIRT1)-mediated autophagy. Dysfunctional mitophagy in response to glucolipotoxicities might play an important role in hepatosteatosis. Here, we investigated the mechanism by which metformin induces mitophagy through restoration of the suppressed Parkin-mediated mitophagy. To this end, our ob/ob mice were divided into three groups: (1) ad libitum feeding of a standard chow diet; (2) intraperitoneal injections of metformin 300 mg/kg; and (3) 3 g/day caloric restriction (CR). HepG2 cells were treated with palmitate (PA) plus high glucose in the absence or presence of metformin. We detected enhanced mitophagy in ob/ob mice treated with metformin or CR, whereas mitochondrial spheroids were observed in mice fed ad libitum. Metabolically stressed ob/ob mice and PA-treated HepG2 cells showed an increase in expression of endoplasmic reticulum (ER) stress markers and cytosolic p53. Cytosolic p53 inhibited mitophagy by disturbing the mitochondrial translocation of Parkin, as demonstrated by immunoprecipitation. However, metformin decreased ER stress and p53 expression, resulting in induction of Parkin-mediated mitophagy. Furthermore, pifithrin-α, a specific inhibitor of p53, increased mitochondrial incorporation into autophagosomes. Taken together, these results indicate that metformin treatment facilitates Parkin-mediated mitophagy rather than mitochondrial spheroid formation by decreasing the inhibitory interaction with cytosolic p53 and increasing degradation of mitofusins.

Keyword: mitochondria

The effect of oleic and on induction of steatosis and cytotoxicity on rat hepatocytes in primary culture.

In vitro models serve as a tool for studies of steatosis. and oleic acids can induce steatosis in cultured hepatocytes. The aim of our study was to verify steatogenic and cytotoxic effects of (PA), oleic (OA) and their combinations as well as their impact on functional capacity of rat primary hepatocytes. Hepatocytes were exposed to OA or PA (0.125-2 mmol/l) or their combination at ratios of 3:1, 2:1 or 1:1 at the final concentrations of 0.5-1 mmol/l. Both OA and PA caused a dose-dependent increase in triacylglycerol content in hepatocytes. PA was more steatogenic at 0.25 and 0.5 mmol/l while OA at 0.75 and 1 mmol/l. PA exhibited a dose-dependent cytotoxic effect associated with ROS production, present markers of apoptosis and necrosis and a decrease in albumin production. OA induced a damage of the cytoplasmic membrane from 1 mM concentration. Mixture of OA and PA induced lower cytotoxicity with less weakened functional capacity than did PA alone. Extent of steatosis was comparable to that after exposure to OA alone. In conclusion, OA or combination of OA with PA is more suitable for simulation of simple steatosis than PA alone.

Keyword: mitochondria

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the -generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: mitochondria

induces ceramide accumulation, mitochondrial protein hyperacetylation, and mitochondrial dysfunction in porcine oocytes.

Low oocyte quality is a possible causal factor of obesity-induced infertility. High (PA) concentration in follicular fluid is a crucial feature noted in obese women. This study examined how high PA concentration reduced mitochondrial quality in oocytes and investigated a possible countermeasure against mitochondrial dysfunction. Cumulus cell-oocyte complexes were obtained from the ovaries of gilts, and incubated in medium containing PA (0.5 mM) or vehicle (BSA) for 44 h. Culturing oocytes at high PA concentration induced mitochondrial dysfunction determined by high reactive oxygen species and low ATP content in oocytes. Furthermore, high PA levels increased mitochondrial acetylation levels determined by a high degree of co-localization of TOMM20 and acetylated-lysine. In addition, high PA levels reduced the expression of Sirtuin 3 (SIRT3) and phosphorylated AMP-activated protein kinase (AMPK), while the AMPK activator, AICAR, restored mitochondrial function as well as oocyte ability and reduced the acetylation of mitochondrial protein. Supplementation of culture medium with dorsomorphin dihydrochloride (an AMPK inhibitor) reduced mitochondrial function and increased mitochondrial protein acetylation. Treatment of oocytes with LB100 (an inhibitor of AMPK dephosphorylation) reduced mitochondrial acetylation levels and restored mitochondrial function. Furthermore, high PA levels increased ceramide accumulation in oocytes, and addition of ceramide to the culture medium also induced mitochondrial dysfunction and increased mitochondrial acetylation. This detrimental effect of ceramide was diminished by AICAR treatment of oocytes. Our results indicated that PA induces ceramide accumulation and downregulates the AMPK/SIRT3 pathway causing mitochondrial protein hyperacetylation and dysfunction in oocytes.

Keyword: mitochondria

Loss of biliverdin reductase-A promotes lipid accumulation and lipotoxicity in mouse proximal tubule cells.

Obesity and increased lipid availability have been implicated in the development and progression of chronic kidney disease. One of the major sites of renal lipid accumulation is in the proximal tubule cells of the kidney, suggesting that these cells may be susceptible to lipotoxicity. We previously demonstrated that loss of hepatic biliverdin reductase A (BVRA) causes fat accumulation in livers of mice on a high-fat diet. To determine the role of BVRA in mouse proximal tubule cells, we generated a CRISPR targeting BVRA for a knockout in mouse proximal tubule cells (BVRA KO). The BVRA KO cells had significantly less metabolic potential and mitochondrial respiration, which was exacerbated by treatment with , a saturated fatty . The BVRA KO cells also showed increased intracellular triglycerides which were associated with higher fatty uptake gene cluster of differentiation 36 as well as increased de novo lipogenesis as measured by higher neutral lipids. Additionally, neutrophil gelatinase-associated lipocalin 1 expression, annexin-V FITC staining, and lactate dehydrogenase assays all demonstrated that BVRA KO cells are more sensitive to -induced lipotoxicity than wild-type cells. Phosphorylation of BAD which plays a role in cell survival pathways, was significantly reduced in -treated BVRA KO cells. These data demonstrate the protective role of BVRA in proximal tubule cells against saturated fatty -induced lipotoxicity and suggest that activating BVRA could provide a benefit in protecting from obesity-induced kidney injury.

Keyword: mitochondria

ANT1-mediated fatty -induced uncoupling as a target for improving myocellular insulin sensitivity.

Dissipating energy via mitochondrial uncoupling has been suggested to contribute to enhanced insulin sensitivity. We hypothesised that skeletal muscle of endurance-trained athletes have increased sensitivity for fatty (FA)-induced uncoupling, which is driven by the mitochondrial protein adenine nucleotide translocase 1 (ANT1).Capacity for FA-induced uncoupling was measured in endurance-trained male athletes (T) and sedentary young men (UT) in an observational study and also in isolated skeletal muscle from Zucker diabetic fatty (ZDF) rats and C2C12 myotubes following small interfering RNA (siRNA)-mediated gene silencing of ANT1. Thus, fuelled by glutamate/succinate (fibres) or pyruvate ( and myotubes) and in the presence of oligomycin to block ATP synthesis, increasing levels of oleate (fibres) or palmitate ( and myotubes) were automatically titrated while respiration was monitored. Insulin sensitivity was measured by hyperinsulinaemic-euglycaemic clamp in humans and via insulin-stimulated glucose uptake in myotubes.Skeletal muscle from the T group displayed increased sensitivity to FA-induced uncoupling (p\u2009=\u20090.011) compared with muscle from the UT group, and this was associated with elevated insulin sensitivity (p\u2009=\u20090.034). ANT1 expression was increased in T (p\u2009=\u20090.013). from ZDF rats displayed decreased sensitivity for FA-induced uncoupling (p\u2009=\u20090.008). This difference disappeared in the presence of the adenine nucleotide translocator inhibitor carboxyatractyloside. Partial knockdown of ANT1 in C2C12 myotubes decreased sensitivity to the FA-induced uncoupling (p\u2009=\u20090.008) and insulin-stimulated glucose uptake (p\u2009=\u20090.025) compared with controls.Increased sensitivity to FA-induced uncoupling is associated with enhanced insulin sensitivity and is affected by ANT1 activity in skeletal muscle. FA-induced mitochondrial uncoupling may help to preserve insulin sensitivity in the face of a high supply of FAs.www.trialregister.nl NTR2002.

Keyword: mitochondria

Berberine attenuates sodium palmitate-induced lipid accumulation, oxidative stress and apoptosis in grass carp(Ctenopharyngodon idella)hepatocyte in vitro.

The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, lipid accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in lipid accumulation-hepatocytes induced by sodium palmitate in vitro. The lipid accumulation-hepatocytes (induced by 0.5\u202fmM sodium palmitate for 24\u202fh) were treated with 5\u202fμM berberine for 12\u202fh. Then, the Cell viability, intracellular triglyceride (TG) content, lipid peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, lipid droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from into the cytoplasm. Beberine could improve intracellular lipid droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.Copyright © 2019. Published by Elsevier Ltd.

Keyword: mitochondria

Characterization of Acyl-CoA synthetase isoforms in pancreatic beta cells: Gene silencing shows participation of ACSL3 and ACSL4 in insulin secretion.

Long-chain acyl-CoA synthetases (ACSLs) convert fatty acids to fatty acyl-CoAs to regulate various physiologic processes. We characterized the ACSL isoforms in a cell line of homogeneous rat beta cells (INS-1 832/13\xa0cells) and human pancreatic islets. ACSL4 and ACSL3 proteins were present in the beta cells and human and rat pancreatic islets and concentrated in insulin secretory granules and less in and negligible in other intracellular organelles. ACSL1 and ACSL6 proteins were not seen in INS-1 832/13\xa0cells or pancreatic islets. ACSL5 protein was seen only in INS-1 832/13\xa0cells. With shRNA-mediated gene silencing we developed stable ACSL knockdown cell lines from INS-1 832/13\xa0cells. Glucose-stimulated insulin release was inhibited ∼50% with ACSL4 and ACSL3 knockdown and unaffected in cell lines with knockdown of ACSL5, ACLS6 and ACSL1. Lentivirus shRNA-mediated gene silencing of ACSL4 and ACSL3 in human pancreatic islets inhibited glucose-stimulated insulin release. ACSL4 and ACSL3 knockdown cells showed inhibition of ACSL enzyme activity more with arachidonate than with palmitate as a substrate, consistent with their preference for unsaturated fatty acids as substrates. ACSL4 knockdown changed the patterns of fatty acids in phosphatidylserines and phosphatidylethanolamines. The results show the involvement of ACLS4 and ACLS3 in insulin secretion.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 diabetes. Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and lipid metabolism. Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: mitochondria

DGAT1-Dependent Lipid Droplet Biogenesis Protects Mitochondrial Function during Starvation-Induced Autophagy.

Lipid droplets (LDs) provide an "on-demand" source of fatty acids (FAs) that can be mobilized in response to fluctuations in nutrient abundance. Surprisingly, the amount of LDs increases during prolonged periods of nutrient deprivation. Why cells store FAs in LDs during an energy crisis is unknown. Our data demonstrate that mTORC1-regulated autophagy is necessary and sufficient for starvation-induced LD biogenesis. The ER-resident diacylglycerol acyltransferase 1 (DGAT1) selectively channels autophagy-liberated FAs into new, clustered LDs that are in close proximity to and are lipolytically degraded. However, LDs are not required for FA delivery to but instead function to prevent acylcarnitine accumulation and lipotoxic dysregulation of . Our data support a model in which LDs provide a lipid buffering system that sequesters FAs released during the autophagic degradation of membranous organelles, reducing lipotoxicity. These findings reveal an unrecognized aspect of the cellular adaptive response to starvation, mediated by LDs.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

High-Fat Diet-Induced Lysosomal Dysfunction and Impaired Autophagic Flux Contribute to Lipotoxicity in the Kidney.

Excessive fat intake contributes to the progression of metabolic diseases cellular injury and inflammation, a process termed lipotoxicity. Here, we investigated the role of lysosomal dysfunction and impaired autophagic flux in the pathogenesis of lipotoxicity in the kidney. In mice, a high-fat diet (HFD) resulted in an accumulation of phospholipids in enlarged lysosomes within kidney proximal tubular cells (PTCs). In isolated PTCs treated with , autophagic degradation activity progressively stagnated in association with impaired lysosomal acidification and excessive lipid accumulation. Pulse-chase experiments revealed that the accumulated lipids originated from cellular membranes. In mice with induced PTC-specific ablation of autophagy, PTCs of HFD-mice exhibited greater accumulation of ubiquitin-positive protein aggregates normally removed by autophagy than did PTCs of mice fed a normal diet. Furthermore, HFD-mice had no capacity to augment autophagic activity upon another pathologic stress. Autophagy ablation also exaggerated HFD-induced mitochondrial dysfunction and inflammasome activation. Moreover, renal ischemia-reperfusion induced greater injury in HFD-mice than in mice fed a normal diet, and ablation of autophagy further exacerbated this effect. Finally, we detected similarly enhanced phospholipid accumulation in enlarged lysosomes and impaired autophagic flux in the kidneys of obese patients compared with nonobese patients. These findings provide key insights regarding the pathophysiology of lipotoxicity in the kidney and clues to a novel treatment for obesity-related kidney diseases.Copyright © 2017 by the American Society of Nephrology.

Keyword: mitochondria

Attenuated lipotoxicity and apoptosis is linked to exogenous and endogenous augmenter of liver regeneration by different pathways.

Nonalcoholic fatty liver disease (NAFLD) covers a spectrum from simple steatosis to nonalcoholic steatohepatitis (NASH) and cirrhosis. Free fatty acids (FFA) induce steatosis and lipo-toxicity and correlate with severity of NAFLD. In this study we aimed to investigate the role of exogenous and endogenous ALR (augmenter of liver regeneration) for FFA induced ER (endoplasmatic reticulum) -stress and lipoapoptosis. Primary human hepatocytes or hepatoma cells either treated with recombinant human ALR (rhALR, 15kDa) or expressing short form ALR (sfALR, 15kDa) were incubated with (PA) and analyzed for lipo-toxicity, -apoptosis, activation of ER-stress response pathways, triacylglycerides (TAG), mRNA and protein expression of lipid metabolizing genes. Both, exogenous rhALR and cytosolic sfALR reduced PA induced caspase 3 activity and Bax protein expression and therefore lipotoxicity. Endogenous sfALR but not rhALR treatment lowered TAG levels, diminished activation of ER-stress mediators C-Jun N-terminal kinase (JNK), X-box binding protein-1 (XBP1) and proapoptotic transcription factor C/EBP-homologous protein (CHOP), and reduced death receptor 5 protein expression. Cellular ALR exerts its lipid lowering and anti-apoptotic actions by enhancing FABP1, which binds toxic FFA, increasing mitochondrial β-oxidation by elevating the mitochondrial FFA transporter CPT1α, and decreasing ELOVL6, which delivers toxic FFA metabolites. We found reduced hepatic mRNA levels of ALR in a high fat diet mouse model, and of ALR and FOXA2, a transcription factor inducing ALR expression, in human steatotic as well as NASH liver samples, which may explain increased lipid deposition and reduced β-oxidation in NASH patients. Present study shows that exogenous and endogenous ALR reduce PA induced lipoapoptosis. Furthermore, cytosolic sfALR changes mRNA and protein expression of genes regulating lipid metabolism, reduces ER-stress finally impeding progression of NASH.

Keyword: mitochondria

ACSL5 genotype influence on fatty metabolism: a cellular, tissue, and whole-body study.

Acyl-CoA Synthetase Long Chain 5 (ACSL5) gene\'s rs2419621 T/C polymorphism was associated with ACSL5 mRNA expression and response to lifestyle interventions. However, the mechanistic understanding of the increased response in T allele carriers is lacking. Study objectives were to investigate the effect of rs2419621 genotype and ACSL5 human protein isoforms on fatty oxidation and respiration.Human ACSL5 overexpression in C2C12 mouse myoblasts was conducted to measure C oxidation and protein isoform localization in vitro. C oxidation studies and Western blot analysis of ACSL5 proteins were carried out in rectus abdominis primary myotubes from 5 rs2419621 T allele carriers and 4 non-carriers. In addition, mitochondrial high-resolution respirometry was conducted on vastus lateralis muscle biopsies from 4 rs2419621 T allele carriers and 4 non-carriers. Multiple linear regression analysis was conducted to test the association between rs2419621 genotype and respiratory quotient related pre- and post-lifestyle intervention measurements in postmenopausal women with overweight or obesity.In comparison to rs2419621 non-carriers, T allele carriers displayed higher levels of i) 683aa ACSL5 isoform, localized mainly in the , playing a greater role in fatty oxidation in comparison to the 739aa protein isoform ii) in vitro CO production in rectus abdominis primary myotubes iii) in vivo fatty oxidation and lower carbohydrate oxidation post-intervention iv) ex vivo complex I and II tissue respiration in vastus lateralis muscle.These results support the conclusion that rs2419621 T allele carriers, are more responsive to lifestyle interventions partly due to an increase in the short ACSL5 protein isoform, increasing cellular, tissue and whole-body fatty utilization. With the increasing effort to develop personalized medicine to combat obesity, our findings provide additional insight into genotypes that can significantly affect whole body metabolism and response to lifestyle interventions.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High-Fat-Diet-Induced Cardiac Dysfunction and Insulin Resistance by Elevating Tafazzin in Mouse Hearts.

High fat (HF)-diet-induced insulin resistance is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac insulin signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low fat (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs insulin signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the . Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores insulin sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced insulin resistance further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to insulin resistance via mediating Drp-1 translocation to the , and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates insulin resistance.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: mitochondria

Crucial Roles of 5-HT and 5-HT2 Receptor in Diabetes-Related Lipid Accumulation and Pro-Inflammatory Cytokine Generation in Hepatocytes.

Previously, we confirmed that liver-synthesized 5-HT rather than non-liver 5-HT, acting on the 5-HT2 receptor (5-HT2R), modulates lipid-induced excessive lipid synthesis (ELS). Here, we further revealed the effects of the hepatocellular 5-HT system in diabetes-related disorders.Studies were conducted in male ICR mice, human HepG2 cells, and primary mouse hepatocytes (PMHs) under gene or chemical inhibition of the 5-HT system, key lipid metabolism, and inflammation-related factors. Protein and messenger RNA expression and levels of the factors were determined via western blotting, reverse transcription PCR, and quantitative assay kits, respectively. Hepatic steatosis with inflammation and fibrosis, intracellular lipid droplet accumulation (LDA), and reactive oxygen species (ROS) location were determined via hematoxylin and eosin, Masson\'s trichrome, Oil red O, and fluorescent-specific staining, respectively. induced the activation of the 5-HT system: the activation of 5-HT2R, primarily 5-HT2AR, in addition to upregulating monoamine oxidase A (MAO-A) expression and 5-HT synthesis, by activating the G protein/ phospholipase C pathway modulated PKCε activation, resulting in ELS with LDA; the activation of NF-κB, which mediates the generation of pro-inflammatory cytokines, was primarily due to ROS generation in the induced by MAO-A-catalyzed 5-HT degradation, and secondarily due to the activation of PKCε. These effects of the 5-HT system were also detected in - or high glucose-treated PMHs and regulated multiple inflammatory signaling pathways. In diabetic mice, co-treatment with antagonists of both 5-HT synthesis and 5-HT2R significantly abolished hepatic steatosis, inflammation, and fibrosis as well as hyperglycemia and dyslipidemia.Activation of the hepatocellular 5-HT system plays a crucial role in inducing diabetes-related hepatic dysfunction and is a potential therapeutic target.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: mitochondria

Metabolic Signature of MELAS/Leigh Overlap Syndrome in Patient-specific Induced Pluripotent Stem Cells Model.

Mitochondrial myopathy, Encephalopathy, Lactic Acidosis, Stroke-like episodes/Leigh overlap syndrome (MELAS) is caused by defects in the mitochondrial respiratory chain. It is still largely unknown how these mitochondrial respiratory chain defects affect cellular metabolisms and lead to variable clinical phenotypes. Here, we analyzed metabolic signatures in a cellular model of MELAS/ Leigh overlap syndrome using untargeted gas chromatography coupled to mass spectrometry (GC-MS). .We obtained fibroblasts from a MELAS/Leigh overlap syndrome patient carrying the heteroplasmic m.10191T>C mutation, and generated induced pluripotent stem cells (iPSCs) from these fibroblast. Isogenic iPSC clones carrying two different loads of the heteroplasmic mutation (ND3hig-iPSC, ND3"*w- iPSC-) were subjected to metabolome analysis. Metabolite profiles, which were identified by GC-MS, were analyzed by principal component analysis (PCA).We were able to identify about 40 metabolites in control fibroblasts and iPSCs. Upon comparative metabolome analysis between fibroblasts and iPSCs, lactic and proline were distinct between the two groups. When we compared patient fibroblasts and control fibroblasts, no significant distinct metabolites were found. On the other hand, patient specific iPSC with high mutational load (ND3high_ iPSC) showed a distinct metabolite profile compared with ND3""-iPSC and control-iPSCs. Metabolites that contributed to this distinction were pyruvate, malic , , stearic , and lactic . This metabolomic signature was only seen in the undifferentiated state of iPSCs and was lost upon differentiationThese findings suggest that patient specific iPSC technology is useful to elucidate unique pathogenic metabolic pathways ,6mitochondrial chain diseases.

Keyword: mitochondria

Proteomic changes in oocytes after in vitro maturation in lipotoxic conditions are different from those in cumulus cells.

Maternal lipolytic metabolic disorders result in a lipotoxic microenvironment in the ovarian follicular fluid (FF) which deteriorates oocyte quality. Although cellular stress response mechanisms are well defined in somatic cells, they remain largely unexplored in oocytes, which have distinct organelle structure and nuclear transcription patterns. Here we used shotgun proteomic analyses to study cellular responses of bovine oocytes and cumulus cells (CCs) after in vitro maturation under lipotoxic conditions; in the presence of pathophysiological (PA) concentration as a model. Differentially regulated proteins (DRPs) were mainly localized in the endoplasmic reticulum, and nuclei of CCs and oocytes, however the DRPs and their direction of change were cell-type specific. Proteomic changes in PA-exposed\xa0CCs were predominantly pro-apoptotic unfolded protein responses (UPRs), mitochondrial and metabolic dysfunctions, and apoptotic pathways. This was also functionally confirmed. Interestingly, although the oocytes were enclosed by CCs during PA exposure, elevated cellular stress levels were also evident. However, pro-survival UPRs, redox regulatory and compensatory metabolic mechanisms were prominent despite evidence of mitochondrial dysfunction, oxidative stress, and reduced subsequent embryo development. The data provides a unique insight that enriches the understanding of the cellular stress responses in metabolically-compromised oocytes and forms a fundamental base to identify new targets for fertility treatments as discussed within.

Keyword: mitochondria

Myristic potentiates -induced lipotoxicity and steatohepatitis associated with lipodystrophy by sustaning de novo ceramide synthesis.

(PA) induces hepatocyte apoptosis and fuels de novo ceramide synthesis in the endoplasmic reticulum (ER). Myristic (MA), a free fatty highly abundant in copra/palmist oils, is a predictor of nonalcoholic steatohepatitis (NASH) and stimulates ceramide synthesis. Here we investigated the synergism between MA and PA in ceramide synthesis, ER stress, lipotoxicity and NASH. Unlike PA, MA is not lipotoxic but potentiated PA-mediated lipoapoptosis, ER stress, caspase-3 activation and cytochrome c release in primary mouse hepatocytes (PMH). Moreover, MA kinetically sustained PA-induced total ceramide content by stimulating dehydroceramide desaturase and switched the ceramide profile from decreased to increased ceramide 14:0/ceramide16:0, without changing medium and long-chain ceramide species. PMH were more sensitive to equimolar ceramide14:0/ceramide16:0 exposure, which mimics the outcome of PA plus MA treatment on ceramide homeostasis, than to either ceramide alone. Treatment with myriocin to inhibit ceramide synthesis and tauroursodeoxycholic to prevent ER stress ameliorated PA plus MA induced apoptosis, similar to the protection afforded by the antioxidant BHA, the pan-caspase inhibitor z-VAD-Fmk and JNK inhibition. Moreover, ruthenium red protected PMH against PA and MA-induced cell death. Recapitulating in vitro findings, mice fed a diet enriched in PA plus MA exhibited lipodystrophy, hepatosplenomegaly, increased liver ceramide content and cholesterol levels, ER stress, liver damage, inflammation and fibrosis compared to mice fed diets enriched in PA or MA alone. The deleterious effects of PA plus MA-enriched diet were largely prevented by in vivo myriocin treatment. These findings indicate a causal link between ceramide synthesis and ER stress in lipotoxicity, and imply that the consumption of diets enriched in MA and PA can cause NASH associated with lipodystrophy.

Keyword: mitochondria

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro diabetes research.

The interplay between hyper-glycemia and -lipidemia in diabetes mellitus (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Down-regulation of stearoyl-CoA desaturase-1 increases susceptibility to -induced lipotoxicity in human trophoblast cells.

In early pregnancy, adequate dietary factors are important for the growth of human trophoblast cells, followed by placental development. Although stearoyl-CoA desaturase 1 (SCD1) is expected to relieve (PA)-induced lipotoxicity by regulating diacylglycerol and ceramide, its function is unclear in human trophoblast cells. The aim was to investigate inhibitory effects of SCD1 activity on PA-induced trophoblast cell death. PA induces cell death and inhibits the invasion of human trophoblast cells (HTR8/SVneo). In addition, we demonstrate that SCD1 has a protective role against PA in human trophoblast cells by regulating AKT-mediated signaling pathway and mitochondrial membrane potential. The knockdown of SCD1 enhances the proapoptotic activity of PA in HTR8/SVneo cells. Lastly, we investigated microRNA expression predicted to target SCD1 and diacylglycerol O-acyltransferase 1 (DGAT1) by PA. Collectively, the results suggest potential roles of SCD1 and DGAT1 in alleviating the toxicity of PA and maintaining lipid homeostasis for normal placentation.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: mitochondria

High-Protein Intake during Weight Loss Therapy Eliminates the Weight-Loss-Induced Improvement in Insulin Action in Obese Postmenopausal Women.

High-protein (HP) intake during weight loss (WL) therapy is often recommended because it reduces the loss of lean tissue mass. However, HP intake could\xa0have adverse effects on metabolic function, because\xa0protein ingestion reduces postprandial insulin sensitivity. In this study, we compared the effects of ∼10% WL with a hypocaloric diet containing 0.8\xa0g protein/kg/day and a hypocaloric diet containing 1.2\xa0g protein/kg/day on muscle insulin action in\xa0postmenopausal women with obesity. We found that HP intake reduced the WL-induced decline in lean tissue mass by ∼45%. However, HP intake also prevented the WL-induced improvements in muscle insulin signaling and insulin-stimulated glucose uptake, as well as the WL-induced adaptations in oxidative stress and cell structural biology pathways. Our data demonstrate that the protein content of a WL diet can have profound effects on metabolic function and underscore the importance of considering dietary macronutrient composition during WL therapy for people with obesity.Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

Keyword: mitochondria

Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.

Mitochondrial trafficking plays a central role in dorsal root ganglion (DRG) neuronal cell survival and neurotransmission by transporting from the neuronal cell body throughout the bundles of DRG axons. In type 2 diabetes (T2DM), dyslipidemia and hyperglycemia damage DRG neurons and induce mitochondrial dysfunction; however, the impact of free fatty acids and glucose on mitochondrial trafficking in DRG neurons remains unknown. To evaluate the impact of free fatty acids compared to hyperglycemia on mitochondrial transport, primary adult mouse DRG neuron cultures were treated with physiologic concentrations of palmitate and glucose and assessed for alterations in mitochondrial trafficking, mitochondrial membrane potential, and mitochondrial bioenergetics. Palmitate treatment significantly reduced the number of motile in DRG axons, but physiologic concentrations of glucose did not impair mitochondrial trafficking dynamics. Palmitate-treated DRG neurons also exhibited a reduction in mitochondrial velocity, and impaired mitochondrial trafficking correlated with mitochondrial depolarization in palmitate-treated DRG neurons. Finally, we found differential bioenergetic effects of palmitate and glucose on resting and energetically challenged in DRG neurons. Together, these results suggest that palmitate induces DRG neuron mitochondrial depolarization, inhibiting axonal mitochondrial trafficking and altering mitochondrial bioenergetic capacity.-Rumora, A. E., Lentz, S. I., Hinder, L. M., Jackson, S. W., Valesano, A., Levinson, G. E., Feldman, E. L. Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.© FASEB.

Keyword: mitochondria

Massive endocytosis triggered by surface membrane palmitoylation under mitochondrial control in BHK fibroblasts.

Large Ca transients cause massive endocytosis (MEND) in BHK fibroblasts by nonclassical mechanisms. We present evidence that MEND depends on mitochondrial permeability transition pore (PTP) openings, followed by coenzyme A (CoA) release, acyl CoA synthesis, and membrane protein palmitoylation. MEND is blocked by inhibiting mitochondrial Ca uptake or PTP openings, depleting fatty acids, blocking acyl CoA synthesis, metabolizing CoA, or inhibiting palmitoylation. It is triggered by depolarizing or promoting PTP openings. After mitochondrial MEND blockade, MEND is restored by cytoplasmic acyl CoA or CoA. MEND is blocked by siRNA knockdown of the plasmalemmal acyl transferase, DHHC5. When acyl CoA is abundant, transient H2O2 oxidative stress or PKC activation initiates MEND, but the immediate presence of H2O2 prevents MEND. The PTP inhibitor, NIM811, significantly increases plasmalemma in normally growing cells. Thus, the MEND pathway may contribute to constitutive as well as pathological plasmalemma turnover in dependence on mitochondrial stress signaling. DOI: http://dx.doi.org/10.7554/eLife.01293.001.

Keyword: mitochondria

Myricanol modulates skeletal muscle-adipose tissue crosstalk to alleviate high-fat diet-induced obesity and insulin resistance.

Skeletal muscle is the predominant site for glucose disposal and fatty consumption. Emerging evidence indicates that the crosstalk between adipose tissue and skeletal muscle is critical in maintaining insulin sensitivity and lipid homeostasis. The current study was designed to investigate whether myricanol improves insulin sensitivity and alleviates adiposity through modulating skeletal muscle-adipose tissue crosstalk.The therapeutic effect of myricanol was evaluated on (PA)-treated C2C12 myotubes and high-fat diet (HFD)-fed mice. The crosstalk between myotubes and adipocytes was evaluated using Transwell assay. The cellular lipid content was examined by Nile red staining. The mitochondrial content was assessed by MitoTracker Green staining and citrate synthase activity, and the mitochondrial function was examined by Seahorse assay. Expression of mitochondria-related and insulin signalling pathway proteins was analysed by Western blot, and the irisin level was determined by elisa kit.Myricanol increased mitochondrial quantity and function through activating AMP-activated protein kinase, resulting in reduced lipid accumulation and enhanced insulin-stimulated glucose uptake, in PA-treated C2C12 myotubes. Furthermore, myricanol stimulated irisin production and secretion from myotubes to reduce lipid content in 3T3-L1 adipocytes. In HFD-fed mice, myricanol treatment alleviated adiposity and insulin resistance through enhancing lipid utilization and irisin production in skeletal muscle and inducing of inguinal fat.Myricanol modulates skeletal muscle-adipose tissue crosstalk, to stimulate of adipose tissue and improve insulin sensitivity in skeletal muscle. Myricanol might be a potential candidate for treating insulin resistance and obesity.© 2019 The British Pharmacological Society.

Keyword: mitochondria

[Insulin: initiation of pool of insulin-dependent cells, targeted transfer of triglycerides and increase of kinetic parameters of oxidation of fatty acids].

The insulin, to provide with energy the biological function of locomotion, formed: a) pool of phylogenetically late insulin-dependent cells; b) highly productive vector variant of transfer of saturated and mono unsaturated fatty acids only to insulin-dependent cells; c) new variant of active absorption of substrates for acquiring energy by cells--apoE/B-100-receptor endocytosis; d) transformation of all endogenically synthesized saturated fatty in oleic mono saturated fatty and e) replacement of potentially ineffective variant of formation of energy in vivo with potentially high-performance oleic variant of metabolism of substrates for turning out of ATP. The insulin expressed synthesis of apoE glucose carrier 4 and stearyl-KoA-desaturase. These occurrences confirm that syndrome of insulin resistance primarily is the pathology of metabolism of fatty acids and only secondary the pathology metabolism of glucose. The multi-functional fatty cells of visceral areolar tissue and specialized adipocytes of subcutaneous fat depots are phylogenetically, regulatory and functionally different cells. They are formed under development of different biological functions: the first ones under realization of biological function of trophology and second ones under realization of biological function of locomotion. At the level of organism, the mechanisms of hypothalamus-fatty cells feedback are realized by peptide leptin and in case of hypothalamus-adipocytes feedback--peptide adiponectin. The potential possibilities of in synthesis of ATP are high and are conditioned only by amount of substrate of acetyl-KoA. This shortage can be chronic as in cases of disorder of insulin function and variant of metabolism of substrates for acquiring energy by cells. The deficiency of acetyl-KoA can be acute as is the case of diabetic coma when surplus amount of ketonic bodies follows the expressed deficiency of acetyl-KoA formed from glucose and fatty acids. Can the intravenous injection of acetyl-KoA be effective under diabetic ketoacidosic coma?

Keyword: mitochondria

Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis.

Autophagy, which is critical for the proper turnover of organelles such as endoplasmic reticulum and , affects diverse aspects of metabolism, and its dysregulation has been incriminated in various metabolic disorders. However, the role of autophagy of myeloid cells in adipose tissue inflammation and type 2 diabetes has not been addressed. We produced mice with myeloid cell-specific deletion of Atg7 (autophagy-related 7), an essential autophagy gene (Atg7 conditional knockout [cKO] mice). While Atg7 cKO mice were metabolically indistinguishable from control mice, they developed diabetes when bred to ob/w mice (Atg7 cKO-ob/ob mice), accompanied by increases in the crown-like structure, inflammatory cytokine expression and inflammasome activation in adipose tissue. Mφs (macrophages) from Atg7 cKO mice showed significantly higher interleukin 1 β release and inflammasome activation in response to a plus lipopolysaccharide combination. Moreover, a decrease in the NAD(+):NADH ratio and increase in intracellular ROS content after treatment with in combination with lipopolysaccharide were more pronounced in Mφs from Atg7 cKO mice, suggesting that mitochondrial dysfunction in autophagy-deficient Mφs leads to an increase in lipid-induced inflammasome and metabolic deterioration in Atg7 cKO-ob/ob mice. Atg7 cKO mice were more susceptible to experimental colitis, accompanied by increased colonic cytokine expression, T helper 1 skewing and systemic bacterial invasion. These results suggest that autophagy of Mφs is important for the control of inflammasome activation in response to metabolic or extrinsic stress, and autophagy deficiency in Mφs may contribute to the progression of metabolic syndrome associated with lipid injury and colitis.

Keyword: mitochondria

Skeletal muscle miR-34a/SIRT1:AMPK axis is activated in experimental and human non-alcoholic steatohepatitis.

Non-alcoholic fatty liver disease (NAFLD) pathogenesis associates with intramyocellular lipid deposition and mitochondrial dysfunction. microRNAs (miRs), including pro-apoptotic miR-34a, are modulated during disease progression in liver tissue and plasma. We aimed to investigate the functional role of the miR-34a/SIRT1:AMP-activated protein kinase (AMPK) pathway in modulating local mitochondrial dysfunction in the skeletal muscle of human and experimental non-alcoholic steatohepatitis. Muscle biopsies were obtained from morbid obese NAFLD patients undergoing bariatric surgery. C57BL/6N mice were fed different NAFLD-inducing diets and C2C12 muscle cells incubated with (PA) in the presence or absence of an AMPK activator, or upon miR-34a functional modulation. Several muscle miRNAs, including miR-34a, were found increased with human NAFLD progression. Activation of the miR-34a/SIRT1:AMPK pathway, concomitant with impairment in insulin signalling mediators and deregulation of mitochondrial-shaping proteins, was evident in C2C12 cells incubated with PA, as well as in the skeletal muscle of all three diet-induced NAFLD mice models. Functional studies established the association between miR-34a- and PA-induced muscle cell deregulation. Of note, activation of AMPK almost completely prevented miR-34a- and PA-induced cellular stress. In addition, the miR-34a/SIRT1:AMPK pathway and mitochondrial dynamics dysfunction were also found amplified in muscle of human NAFLD. Finally, muscle miR-34a expression and mitofusin 2 (Mfn2) protein levels correlated with hallmarks of NAFLD and disease progression. Our results indicate that activation of the miR-34a/SIRT1:AMPK pathway leads to mitochondrial dynamics dysfunction in skeletal muscle of human and experimental NAFLD, representing an appealing prospective target in metabolic syndrome. KEY MESSAGES: Skeletal muscle microRNAs are modulated during NAFLD progression. -induced muscle cell dysfunction occurs, at least in part, through activation of the miR-34a/SIRT1:AMPK pathway. miR-34a/SIRT1:AMPK activation associates with dynamics dysfunction in human NAFLD.

Keyword: mitochondria

[Hypoglycemic activity of hypolipidemic preparations].

The system of glucose metabolism regulation is millions of years older than the insilin system and locomotor function. For this reason, the hypoglycemic activity of the hormone is mediated through fatty (FA) metabolism. Insulin blocks the ability of to oxidize ketone bodies, short-, medium- and long-chain FA and makes them oxidize glucose, i.e. a physiologically unoptimal substrate. The relationship between FA and glucose in the Rnadle cycle is apparent only on the autocrine level (in the cell); they determine alternation of nutritive function (trophology) and biological exo/endo-reactions (after and without feeding respectively). Most antidiabetic medicines exhibit hypoglycemic activity, like insulin; they reduce the level of lipid substrates of oxidation in cytosol and have to oxidize glucose. In these conditions, insulin increases glucose uptake by the cells mediated through GLUT4 transporters. Sulfonylurea derivatives enhance secretion of insulin by beta-cell. Biguanides covalently and irreversibly bind ketone bodies thereby preventing their oxidation by . Fibrates, glitazones, flavonides, flavones, lipoic thio-FA, endogenous eicosanoids, derivatives of Ω-3 and Ω-6 essential polyenic FA, and conjugated unsaturated FA are agonists of peroxisome proliferator-activated receptors. They stimulate α-, β-, and Ω-oxidation of all exogenous a physiological FA and excessive saturated FA in peroxisomes which leads to cytosol hypolipidemia. Hypoglycemic preparations with the activity of oxidation beta-blockers arrest FA uptake by . Hypoglycemic Ω-3 essential polyenoic FA activate GLUT4 function. Type 2 diabetes in middle-aged patients is a symptom of atherosclerosis, i.e. deficit of essential polyenoic FA caused by disturbed phospholipid synthesis and GLUT4 function. Diabetes should be in the first place considered as pathological FA metabolism and only in the second place as glucose pathology. This inference needs to be taken into account in the treatment of diabetes and in strategic programs of its prophylaxis.

Keyword: mitochondria

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving lipid metabolism, antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis (NASH) is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high fat high fructose (HFHF) diet fed C57BL/6J mice, used as model of NASH. The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved lipid homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, are intracellular source of reactive oxygen species (ROS) and important sites of lipid metabolism, we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti-NASH potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: mitochondria

Mutation of IDH1 aggravates the fatty ‑induced oxidative stress in HCT116 cells by affecting the mitochondrial respiratory chain.

Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the metabolic reprogramming of cancer cells; however their functions in lipid metabolism remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic (OA) or (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive oxygen species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of , immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty ‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.

Keyword: mitochondria

, but not high-glucose, induced myocardial apoptosis is alleviated by N‑acetylcysteine due to attenuated mitochondrial-derived ROS accumulation-induced endoplasmic reticulum stress.

Pharmacological inhibition of reactive oxygen species (ROS) is a potential strategy to prevent diabetes-induced cardiac dysfunction. This study was designed to investigate precise effects of antioxidant N‑acetylcysteine (NAC) in alleviating diabetic cardiomyopathy (DCM). Echocardiography and histologic studies were performed 12 weeks after streptozocin injection. Protein levels involved in endoplasmic reticulum stress (ERS) and apoptosis were analyzed by western blotting in diabetic hearts or high-glucose (HG, 30\u2009mM)- and (PA, 300\u2009μM)-cultured neonatal rat cardiomyocytes (NRCMs). ROS generation and structural alterations of were also assessed. We report that NAC alleviated diabetes-induced cardiac abnormality, including restored ejection fraction (EF %), fraction shortening (FS %), peak E to peak A ratio (E/A) and reduced cardiac hypertrophy and fibrosis. These effects were concomitant with blocked ERS and apoptosis, as evidenced by inactivation of phosphorylated inositol-requiring enzyme-1α (IRE1α)/spliced X-box binding protein 1 (XBP1), phosphorylated protein kinase-like kinase (PERK)/phosphorylated eukaryotic initiation factor 2α (eIF2α) and glucose-regulated protein 78 (GRP78)/activating transcription factor 6 (ATF6α)/C/EBP homologous protein (CHOP) pathways, as well as suppressed Bcl-2-associated X protein (BAX)/B-cell lymphoma-2 (Bcl-2) and cleaved caspase 3 expressions. Mechanistically, PA mediated excessive mitochondrial ROS generation and oxidative stress, which were antagonized by NAC and Mito-TEMPO, a mitochondrial ROS inhibitor. No effects were noted by addition of apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, and NADPH oxidase 4 (NOX 4) and NOX 2 expressions were not altered, indicating that PA-induced ROS generation is independent of NADPH oxidases. Most intriguingly, HG failed to promote ROS production despite its ability to promote ERS and apoptosis in NRCMs. Collectively, these findings indicate that NAC primarily abrogates PA-mediated mitochondrial ROS through ERS and therefore alleviates myocardial apoptosis but has little effect on HG-induced cardiac injury. This uncovers a potential role for NAC in formulating novel cardioprotective strategies in DCM patients.

Keyword: mitochondria

Hepatic FTO expression is increased in NASH and its silencing attenuates -induced lipotoxicity.

Non-alcoholic steatohepatitis (NASH) is one of the most common causes of liver failure worldwide. It is characterized by excess fat accumulation, inflammation, and increased lipotoxicity in hepatocytes. Currently, there are limited treatment options for NASH due to lack of understanding of its molecular etiology. In the present study, we demonstrate that the expression of fat mass and obesity associated gene (FTO) is significantly increased in the livers of NASH patients and in a rodent model of NASH. Furthermore, using human hepatic cells, we show that genetic silencing of FTO protects against palmitate-induced oxidative stress, mitochondrial dysfunction, ER stress, and apoptosis in\xa0vitro. Taken together, our results show that FTO may have a deleterious role in hepatic cells during lipotoxic conditions, and strongly suggest that up-regulation of FTO may contribute to the increased liver damage in NASH.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: mitochondria

Palmitate activation by fatty transport protein 4 as a model system for hepatocellular apoptosis and steatosis.

Fatty transport protein (FATP) 4 is a minor FATP in the liver but it has some activity towards palmitate 16:0 (Pal). We here chose FATP4 as a representative model enzyme for acyl-CoA synthetases (ACSs), and FATPs to determine whether Pal activation would lead to apoptosis and alteration in lipid metabolism. By using FATP4 overexpressed (FATP4) Huh-7 cells, we showed that FATP4 was localized in the endoplasmic reticulum (ER) and of FATP4 cells. FATP4 cells were more responsive to Pal than the control GFP cells in increasing palmitoyl-CoA and oleoyl-CoA activities as well as apoptosis by ~2-3 folds. The lipoapoptosis susceptibility by FATP4 was coupled with the increased JNK, PUMA, caspase3, PARP-1 activation as well as Rac-1-mediated cytoskeletal reorganization, and decreased insulin sensitivity. This was associated with increased contents of neutral lipids and significant alteration in composition of phospholipids and sphingolipids including increased lysophosphatidylcholine (LPC), ceramide, and hexosylceramide, as well as an increase of saturated:polyunsaturated fatty ratio in LPC and PC, but a decrease of this ratio in phosphatidylethanolamine pool. By use of ceramide synthase inhibitors, our results showed that FATP4-sensitized lipoapoptosis was not mediated by ceramides. Moreover, FATP4 expression was increased in fatty livers in vivo. Thus, our model system has provided a clue that Pal activation FATP4 triggers hepatocellular apoptosis via altered phospholipid composition and steatosis by acylation into complex lipids. This may be a redundant mechanism for other ER-localizing ACSs and FATPs in the liver, and hence their involvement in the development of fatty liver disease.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: mitochondria

The novel NADPH oxidase 4 selective inhibitor GLX7013114 counteracts human islet cell death in vitro.

It has been proposed that pancreatic beta-cell dysfunction in type 2 diabetes is promoted by oxidative stress caused by NADPH oxidase (Nox) over-activity. The aim of the present study was to evaluate the efficacy of novel Nox inhibitors as protective agents against cytokine- or high glucose + palmitate-induced human beta-cell death. The Nox2 protein was present mainly in the cytoplasm and was induced by cytokines. Nox4 protein immunoreactivity, with some nuclear accumulation, was observed in human islet cells, and was not affected by islet culture in the presence of cytokines or high glucose + palmitate. Nox inhibitors with partial or no isoform selectivity (DPI, dapsone, GLX351322, and GLX481372) all reduced ROS production of human islet cells exposed to high glucose + palmitate. This was paralleled by improved viability and reduced caspase 3 activation. The Nox1 selective inhibitor ML171 failed to reduce human islet cell death in response to both cytokines and high glucose + palmitate. The selective Nox2 inhibitor Phox-I2 also failed to protect against cytokines, but protected partially against high glucose + palmitate-induced cellular death. The highly selective Nox4 inhibitor GLX7013114 protected islet cells against both cytokines and high glucose + palmitate. However, as no osmotic control for high glucose was used, we cannot exclude the possibility that the high glucose effect was due to osmosis. It is concluded that Nox4 may participate in stress-induced islet cell death in human islets in vitro. We propose that Nox4 mediates pro-apoptotic effects in intact islets under stressful conditions and that selective Nox4-inhibition may be a therapeutic strategy in type 2 diabetes.

Keyword: mitochondria

Docosahexaenoic and Eicosapentaenoic Acids Prevent Altered-Muc2 Secretion Induced by by Alleviating Endoplasmic Reticulum Stress in LS174T Goblet Cells.

Diets high in saturated fatty acids (FA) represent a risk factor for the development of and associated metabolic disorders, partly through their impact on the epithelial cell barrier integrity. We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus barrier. To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or -9, -6, or -3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic nor -9 and -6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 diabetes and or inflammatory bowel disease. These in vitro data remain to be explored in vivo in a context of dietary .

Keyword: obesity

Phytochemistry, Traditional Uses and Pharmacological Profile of Rose Hip: A Review.

The fruit of genus Rosa, known as "rose hip", is frequently used in different traditional medicines. Rose hips have long been used to treat kidney stones, gastroenteric ailments, hypertension and respiratory problems such as bronchitis, cough and cold.This review is focused on the ethnopharmacological uses of rose hip as well as phytochemical and pharmacological aspects.Ethno-medical uses of rose hip have been recorded in many countries since a long time. Approximately, 129 chemical compounds have been isolated and identified from rose hip. This fruit contains some major active components such as flavonoids, tannins, anthocyanin, phenolic compounds, fatty oil, organic acids and inorganic compounds. Scientific studies have suggested a wide range of pharmacological activities for rose hip including antioxidant, anti-inflammatory, anti-, anti-cancer, hepatoprotective, nephroprotective, cardioprotective, antiaging, anti H. pylori, neuroprotective and antinociceptive activities. In particular, the rose hip powder and extract have been reported to exert therapeutic effects on arthritis.Some of the ethnomedical indications of rose hip, such as nephroprotective and gastroproetctive actions, have been confirmed by preclinical pharmacological studies. Additional investigations on the pharmacological effects of rose hip as well as evidence from randomized controlled trials are essential to assess the therapeutic value of this natural product.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: obesity

Polymorphism in the Gene Is Associated to Changes in Fatty Metabolism and Circulating Endocannabinoid Levels Distinctively in Normal Weight and Obese Subjects.

The multifunctional CD36 scavenger receptor facilitates fatty (FA) uptake and oxidation and it has been involved in the pathophysiology related to dysfunctional FA metabolism. The common variant in the gene, (A/G), whose allele A is characterized by a reduced protein expression, has been associated with taste sensitivity to and preference for fat. We therefore aimed at evaluating whether the polymorphism may influence fatty metabolism and endocannabinoid biosynthesis in normal weight (NW) and obese (OB) subjects. Red blood cell (RBC) fatty composition, and plasma endocannabinoid levels were determined. In NW subjects with AA genotype was found a marked reduction of RBC saturated fatty acids and /linoleic ratio (PA/LA), considered as lipogenesis (DNL) biomarkers. Remarkably, to the reduction of DNL biomarkers corresponded an increase of omega-6 index, an indirect marker of the impact on fatty metabolism of dietary omega-6 fatty acids, endocannabinoid levels and a higher waist/hip ratio. The presence of the G allele was instead associated with increased endocannabinoid plasma levels and a trend for increased waist/hip ratio in obese subjects, even though exhibited decreased BMI with respect to those with AA genotype. These data indicate that the polymorphism, , leads to a distinct metabolic pattern in NW and in OB subjects. Therefore, their determination may be crucial in developing personalized therapeutic strategies for ameliorating dyslipidemia and other metabolic disorders.

Keyword: obesity

Curcumin protects against -induced apoptosis via the inhibition of endoplasmic reticulum stress in testicular Leydig cells.

(PA) is a common saturated fatty that induces apoptosis in various types of cells, including testicular Leydig cells. There is evidence suggesting that PA is increased in patients with and that PA-induced cell apoptosis may play an important role in -related male infertility. Curcumin, a natural polyphenol, has been reported to exert cytoprotective effects in various cell types. However, the cytoprotective effect of curcumin against PA-induced apoptosis in Leydig cells remains unknown. Therefore, the current study was performed to investigate the protective effects of curcumin in response to PA-induced toxicity and apoptosis in murine Leydig tumor cell line 1 (MLTC-1) cells and explore the mechanism underlying its anti-apoptotic action.MLTC-1 cells were cultured in Roswell Park Institute-1640 medium and divided into five groups. First four groups were treated with 50-400\u2009μM PA, 400\u2009μM PA\u2009+\u20095-40\u2009μM curcumin, 400\u2009μM PA\u2009+\u2009500\u2009nM 4-phenylbutyric (4-PBA, an endoplasmic reticulum (ER) stress inhibitor), and 500\u2009nM thapsigargin (TG, an ER stress inducer)\u2009+\u200920\u2009μM curcumin, respectively, followed by incubation for 24\u2009h. Effects of PA and/or curcumin on viability, apoptosis, and ER stress in MLTC-1 cells were then determined by cell proliferation assay, flow cytometry, and western blot analysis. The fifth group of MLTC-1 cells was exposed to 400\u2009μM of PA and 5\u2009IU/mL of human chorionic gonadotropin (hCG) for 24\u2009h in the absence and presence of curcumin, followed by measurement of testosterone levels in cell-culture supernatants by enzyme-linked immunosorbent assay (ELISA). Rats fed a high-fat diet (HFD) were treated with or without curcumin for 4\u2009weeks, and the testosterone levels were detected by ELISA.Exposure to 100-400\u2009μM PA reduced cell viability, activated caspase 3, and enhanced the expression levels of the apoptosis-related protein BCL-2-associated X protein (BAX) and ER stress markers glucose-regulated protein 78 (GRP78) and CCAAT/enhancer binding protein homologous protein (CHOP) in MLTC-1 cells. Treating cells with 500\u2009nM 4-PBA significantly attenuated PA-induced cytotoxicity through inhibition of ER stress. Curcumin (20\u2009μM) significantly suppressed PA- or TG-induced decrease in cell viability, caspase 3 activity, and the expression levels of BAX, CHOP, and GRP78. In addition, treating MLTC-1 cells with 20\u2009μM curcumin effectively restored testosterone levels, which were reduced in response to PA exposure. Similarly, curcumin treatment ameliorated the HFD-induced decrease in serum testosterone level in vivo.The present study suggests that PA induces apoptosis via ER stress and curcumin ameliorates PA-induced apoptosis by inhibiting ER stress in MLTC-1 cells. This study suggests the application of curcumin as a potential therapeutic agent for the treatment of -related male infertility.

Keyword: obesity

Compound C Protects Mice from HFD-Induced and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to lipid metabolism and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: obesity

Does the enterolactone (ENL) affect fatty transporters and lipid metabolism in liver?

NAFLD as a result of inappropriate diet and , may progress to sever conditions such as: type 2 diabetes mellitus or steatohepatitis, and has recently become a prevalent topic of numerous investigations. Due to its dangerous aftermaths, finding new substances, such as polyphenols and their derivatives, which might reduce liver steatosis is the main target of research into NAFLD treatment. Hence, the aim of the present study was to evaluate the effect(s) of enterolactone (ENL), a metabolite of secoisolariciresinol (SECO), on lipid metabolism together with changes in the expression of fatty transporters in fatty liver.The experiments were conducted on HepG2 cells incubated with either ENL and/or during 16\xa0h exposure. The expression of selected fatty transport proteins: FATP2, FATP5, CD36, FABPpm, ABCA1, MTP, ACBP and L-FABP, as well as the proteins directly involved in lipogenesis (FAS), oxidation pathway (CPT 1), and lipid metabolism (PPARα, LXR, SREBP1c, pAMPK) was estimated by Western Blot. Intra and extracellular lipid contents were assessed by Gas-Liquid Chromatography. The data was analyzed with two-way analysis of variance (ANOVA), and results were considered to be statistically significant at \xa0≤\xa00.05.ENL stimulated extracellular efflux of free fatty acids (FFA) and triacylglicerols (TAG) to the medium, while, it had no influence on FATP-family mediated intracellular fatty uptake. Moreover, ENL decreased the expression of CPT 1, pAMPK, PPARα, increased SREBP1c and had no effect on LXR, and FAS content.The findings of our study demonstrate that ENL had opposite effect on liver steatosis in comparison with other polyphenols what suggests that it may be an inactive metabolite. ENL did not affect significantly the intracellular accumulation of FFA, DAG and TAG, yet it promoted their extracellular efflux. Furthermore, it inhibited ß-oxydation and intracellular lipid metabolism what may contribute to the progression of NAFLD.

Keyword: obesity

Preliminary evidence for reduced adipose tissue inflammation in vegetarians compared with omnivores.

There are links between and inflammation that may relate activation of pro-inflammatory pathways by dietary factors. Because dietary fat intake of vegetarians is thought to be more beneficial than that of omnivores, we hypothesized that obese vegetarians would have less adipose tissue inflammation and lower intramyocellular ceramide concentrations than equally obese omnivores.Eight obese vegetarian (1 male) and 8 obese omnivore volunteers (1 male) completed a Food Frequency Questionnaire, underwent body composition measures, subcutaneous adipose tissue and muscle biopsies. We used immunohistochemistry to measure adipose macrophage (ATM) and senescent cells. Plasma free fatty (FFA), adipose FA and muscle ceramide profiles were measured using liquid chromatography/mass spectrometry. Student t tests were used for the comparison of primary outcomes; univariate regression analysis was used to test for associations between dietary patterns and ATMs (secondary analysis).There were no differences in age (38\u2009±\u20098 vs. 39\u2009±\u20098\u2009years), BMI (32.2\u2009±\u20092.6 vs. 33.3\u2009±\u20091.9\u2009kg/m) or percent body fat (44\u2009±\u20098 vs. 45\u2009±\u20094) between the vegetarians and omnivores. Vegetarians consumed 42% (P\u2009=\u20090.02) less saturated fat and 50% (P\u2009=\u20090.04) less cholesterol than the omnivores. Plasma FFA of vegetarians had lesser proportions of (24\u2009±\u20093 vs. 29\u2009±\u20094%, P\u2009=\u20090.02) and vegetarians had fewer femoral pro-inflammatory ATMs than omnivores (3.6\u2009±\u20092.8 vs. 7.9\u2009±\u20094.4 per 100 adipocytes, respectively; P\u2009=\u20090.02). Omnivores had 50% greater (P\u2009=\u20090.01) expression of TNF mRNA in abdominal fat. We found no significant between group differences in muscle ceramide concentrations.Although the sample size is small, these results may indicate that dietary patterns play a role in adipose tissue inflammation, as reflected by reduced number of femoral ATMs in obese vegetarians than obese omnivores.

Keyword: obesity

causes insulin resistance in granulosa cells via activation of JNK.

is a worldwide health problem with rising incidence and results in reproductive difficulties. Elevated saturated free fatty acids (FFAs) in can cause insulin resistance (IR) in peripheral tissues. The high intra-follicular saturated FFAs may also account for IR in ovarian granulosa cells (GCs). In the present study, we investigated the relationship between saturated FFAs and IR in GCs by the use of (PA). We demonstrated that the glucose uptake in cultured GCs and lactate accumulation in the culture medium were stimulated by insulin, but the effects of insulin were attenuated by PA treatment. Besides, insulin-induced phosphorylation of Akt was reduced by PA in a dose and time-dependent manner. Furthermore, PA increased phosphorylation of JNK and JNK blockage rescued the phosphorylation of Akt which was down-regulated by PA. These findings highlighted the negative effect of PA on GCs metabolism and may partially account for the -related reproductive disorders.

Keyword: obesity

Scopoletin intervention in pancreatic endoplasmic reticulum stress induced by lipotoxicity.

Endoplasmic reticulum (ER), a dynamic organelle, plays an essential role in organizing the signaling pathways involved in cellular adaptation, resilience, and survival. Impairment in the functions of ER occurs in a variety of nutritive disorders including and type 2 diabetes. Here, we hypothesize that (scopoletin) SPL, a coumarin, has the potential to alleviate ER stress induced in vitro and in vivo models by lipotoxicity. To test this hypothesis, the ability of SPL to restore the levels of proteins of ER stress was analyzed. Rat insulinoma 5f (RIN5f) cells and Sprague Dawley rats were the models used for this study. Groups of control and high-fat, high-fructose diet (HFFD)-fed rats were treated with either SPL or 4-phenylbutyric . Status of ER stress was enumerated by quantitative RT-PCR, Western blot, electron microscopic, and immunohistochemical studies. Proximal proteins of ER stress inositol requiring enzyme 1 (IRE1), protein kinase like endoplasmic reticulum kinase (PERK), and activating transcription factor 6 (ATF6) were reduced in the β-cells by SPL. The subsequent signaling proteins X-box binding protein 1, eukaryotic initiation factor2α, activating transcription factor 4, and C/EBP homologous protein were also suppressed in their expression levels when treated with SPL. IRE1, PERK signaling leads to c-Jun-N-terminal kinases phosphorylation, a kinase that interrupts insulin signaling, which was also reverted upon scopoletin treatment. Finally, we confirm that SPL has the ability to suppress the stress proteins and limit pancreatic ER stress which might help in delaying the progression of insulin resistance.

Keyword: obesity

The Mitochondria-Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

Mitochondria-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive oxygen species (ROS) production during glucose metabolism in β cells can be exacerbated under hyperglycaemic conditions such as type 2 diabetes (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: obesity

Glucagon-like peptide-1 analog prevents -related glomerulopathy by inhibiting excessive autophagy in podocytes.

To investigate the role of glucagon-like peptide-1 analog (GLP-1) in high-fat diet-induced -related glomerulopathy (ORG). Male C57BL/6 mice fed a high-fat diet for 12 wk were treated with GLP-1 (200 μg/kg) or 0.9% saline for 4 wk. Fasting blood glucose and insulin and the expression of podocin, nephrin, phosphoinositide 3-kinase (PI3K), glucose transporter type (Glut4), and microtubule-associated protein 1A/1B-light chain 3 (LC3) were assayed. Glomerular morphology and podocyte foot structure were evaluated by periodic -Schiff staining and electron microscopy. Podocytes were treated with 150 nM GLP-1 and incubated with 400 μM (PA) for 12 h. The effect on autophagy was assessed by podocyte-specific Glut4 siRNA. Insulin resistance and autophagy were assayed by immunofluorescence and Western blotting. The high-fat diet resulted in weight gain, ectopic glomerular lipid accumulation, increased insulin resistance, and fusion of podophyte foot processes. The decreased translocation of Glut4 to the plasma membrane and excess autophagy seen in mice fed a high-fat diet and in PA-treated cultured podocytes were attenuated by GLP-1. Podocyte-specific Glut4 siRNA promoted autophagy, and rapamycin-enhanced autophagy worsened the podocyte injury caused by PA. Excess autophagy in podocytes was induced by inhibition of Glut4 translocation to the plasma membrane and was involved in the pathology of ORG. GLP-1 restored insulin sensitivity and ameliorated renal injury by decreasing the level of autophagy.

Keyword: obesity

GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic fatty homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced liver dysfunction, hepatic steatosis and insulin resistance. In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and lipogenesis, and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), fatty synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting lipogenesis and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the liver may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

Levels of ester of hydroxystearic (PAHSA) are reduced in the breast milk of obese mothers.

To achieve optimal development of a newborn, breastfeeding is extensively recommended, but little is known about the role of non-nutritive bioactive milk components. We aimed to characterize the fatty esters of hydroxy fatty acids (FAHFAs), namely hydroxystearic acids (PAHSAs)-endogenous lipids with anti-inflammatory and anti-diabetic properties, in human breast milk. Breast milk samples from 30 lean (BMI=19-23) and 23 obese (BMI>30) women were collected 72h postpartum. Adipose tissue and milk samples were harvested from C57BL/6J mice. FAHFA lipid profiles were measured using reverse phase and chiral liquid chromatography-mass spectrometry method. PAHSA regioisomers as well as other FAHFAs were present in both human and murine milk. Unexpectedly, the levels of 5-PAHSA were higher relative to other regioisomers. The separation of both regioisomers and enantiomers of PAHSAs revealed that both R- and S-enantiomers were present in the biological samples, and that the majority of the 5-PAHSA signal is of R configuration. Total PAHSA levels were positively associated with weight gain during pregnancy, and 5-PAHSA as well as total PAHSA levels were significantly lower in the milk of the obese compared to the lean mothers. Our results document for the first time the presence of lipid mediators from the FAHFA family in breast milk, while giving an insight into the stereochemistry of PAHSAs. They also indicate the negative effect of on 5-PAHSA levels. Future studies will be needed to explore the role and mechanism of action of FAHFAs in breast milk.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: obesity

The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis.

Hyperglycemia and hyperlipidemia (glycolipotoxicity)-triggered islet β-cell dysfunction is known to drive the progression of -related type 2 diabetes, however the underlying mechanisms have not been clearly elucidated. The current study aimed to investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP-5) in islet cells under glucolipotoxic conditions. Using gene overexpression and knockdown approaches, we demonstrated that MKP-5 could alleviate glucolipotoxicity-induced apoptosis via the endoplasmic reticulum (ER) stress and mitochondrial apoptosis pathways owing to the altered regulation of caspase family members and ER stress-related molecules in MIN6 and primary islet cells. Overexpression of MKP-5 reversed the glucose and (GP)-induced impairment of insulin secretion as well as the abnormal decreases in the expression of islet functional genes, thereby maintaining the normal insulin secretory functionality, whereas the absence of MKP-5 aggravated islet cell dysfunction. In parallel, the production of ROS and increased inflammation-associated genes in response to GP were also reduced upon MKP-5 overexpression. Further, inhibition of JNK or P38 MAPK pathways resisted to glucolipotoxicity observed in MKP-5 knockdown MIN6 cells. These findings indicate that MKP-5 is an important mediator for glucolipotoxicity-induced islet cell dysfunction and apoptosis, with JNK and P38 as the critical downstream pathways.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: obesity

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: obesity

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 diabetes.

Metabolic stress-induced low-grade chronic inflammation plays an important role in the development of insulin-resistance and type 2 diabetes (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the -induced metabolic stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: obesity

Heat shock protein 70 promotes lipogenesis in HepG2 cells.

The increasing prevalence of non-alcoholic fatty liver disease (NAFLD) has followed the international rise in rates. Multiple mechanisms are involved in NAFLD, including endoplasmic reticulum stress and oxidative stress. Heat shock protein 70 (HSP70), which is abundant in most organisms, is sensitive to stress. However, the role of HSP70 in NAFLD has not been investigated. Here, we investigated the possible role of HSP70 in lipid synthesis.C57BL/6 mice were fed a high-fat diet, and HepG2 cells were treated with 0.5\xa0mM (PA). HSP70 expression was detected by qPCR, Western blot and immunohistochemistry. Total cholesterol (TC) and triglyceride (TG) levels were detected by enzyme-linked immunosorbent assay (ELISA). After Hsp70 overexpression and knockdown, TC and TG levels and FAS, SCD, and ACC expression were detected.HSP70 expression was significantly increased in the livers of obese mice. In vitro, HSP70 expression was markedly induced by PA in HepG2 cells. Notably, HSP70 overexpression in HepG2 cells enhanced TC and TG synthesis, in parallel with the upregulation of lipogenic genes, including FAS, SCD and ACC. By contrast, HSP70 knockdown decreased the levels of cellular lipids and the expression of FAS, SCD, and ACC in HepG2 cells. Together, our results suggest that HSP70 may promote lipogenesis in HepG2 cells.Heat shock protein 70 promotes lipogenesis in HepG2 cells.

Keyword: obesity

Nicotinamide riboside regulates inflammation and mitochondrial markers in AML12 hepatocytes.

The NAD precursor nicotinamide riboside (NR) is a type of vitamin B found in cow\'s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced , Alzheimer\'s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes.A subset of hepatocytes was treated with (PA; 250 µM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 µM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed.Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes.These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.

Keyword: obesity

Mitochondria-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.

Can we use a mitochondrial-targeted antioxidant (Mitoquinone) during in vitro embryo culture to rescue developmental competence of oocytes matured under lipotoxic conditions, exhibiting mitochondrial dysfunction and oxidative stress?Supplementation of embryo culture media with Mitoquinone reduced oxidative stress and prevented mitochondrial uncoupling in embryos derived from metabolically compromised oocytes in vitro, leading to higher blastocyst rates and lower blastomeric apoptosis.Maternal metabolic disorders, such as and type-II diabetes are associated with hyperlipidemia and elevated free fatty (FFA) concentrations in the ovarian follicular fluid (FF). Oocyte maturation under these lipotoxic conditions results in increased oxidative stress levels, mitochondrial dysfunction, reduced developmental competence and disappointing IVF results.A well-described bovine oocyte IVM model was used, where a pathophysiologically relevant elevated FF concentrations of (PA; 150\xa0μM or 300\xa0μM) were added to induce oxidative stress. After fertilization (Day 0, D0), zygotes were in vitro cultured (IVC, from D1 to D8) in standard fatty -free media in the presence or absence of Mitoquinone or its carrier triphenyl-phosphonium.Embryo cleavage and fragmentation (D2) and blastocyst rates (D8) were recorded. Mitochondrial activity and oxidative stress in cleaved embryos at D2 were determined using specific fluorogenic probes and confocal microscopy. D8 blastocysts were used to (i) examine the expression of marker genes related to mitochondrial unfolded protein responses (UPRmt; HSPD1 and HSPE1), mitochondrial biogenesis (TFAM), endoplasmic reticulum (ER) UPR (ATF4, ATF6 and BiP) and oxidative stress (CAT, GPX1 and SOD2) using real time RT-PCR; (ii) determine cell differentiation and apoptosis using CDX-2 and cleaved caspase-3 immunostaining; and (iii) measure mtDNA copy numbers. This was tested in a series of experiments with at least three independent replicates for each, using a total of 2525 oocytes. Differences were considered significant if a P value was <0.05 after Bonferroni correction.Exposure to PA during IVM followed by culture under control conditions resulted in a significant increase in oxidative stress in embryos at D2. This was associated with a significant reduction in mitochondrial inner membrane potential (uncoupling) compared with solvent control (P\xa0<\u20090.05). The magnitude of these effects was PA-concentration dependent. Consequently, development to the blastocysts stage was significantly hampered. Surviving blastocysts exhibited high apoptotic cell indices and upregulated mRNA expression indicating persistent oxidative stress, mitochondrial and ER UPRs. In contrast, supplementation of PA-derived zygotes with Mitoquinone during IVC (i) prevented mitochondrial uncoupling and alleviated oxidative stress at D2; and (ii) rescued blastocyst quality; normalized oxidative stress and UPR related genes and apoptotic cell indices (P\xa0>\u20090.01 compared with solvent control). Mitoquinone also improved blastocyst rate in PA-exposed groups, an effect that was dependent on PA concentration.N/A.This is a fundamental study performed using a bovine in vitro model using PA-induced lipotoxicity during oocyte maturation. PA is the most predominant FFA in the FF that is known to induce lipotoxicity; however, in vivo maturation in patients suffering from maternal metabolic disorders involve more factors that cannot be represented in one model. Nevertheless, focusing on the carryover oxidative stress as a known key factor affecting developmental competence, and considering the novel beneficial rescuing effects of Mitoquinone shown here, we believe this model is of high biological relevance.Human oocytes collected for IVF treatments from patients with maternal metabolic disorders are vulnerable to lipotoxicity and oxidative stress during in vivo maturation. The results shown here suggest that mitochondrial targeted therapy, such as using Mitoquinone, during IVC may rescue the developmental competence and quality of these compromised oocytes. After further clinical trials, this may be a valuable approach to increase IVF success rates for infertile patients experiencing metabolic disorders.This study was financially supported by a BOF/KP grant number 34399, from the University of Antwerp, Belgium. W.F.A.M. was supported by a postdoctoral fellowship from the Research Foundation-Flanders (FWO), grant number 12I1417N, Antwerp, Belgium. The Leica SP 8 confocal microscope used in this study was funded by the Hercules Foundation of the Flemish Government (Hercules grant AUHA.15.12). All authors have no financial or non-financial competing interests to declare.© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: obesity

Implication of Free Fatty Acids in Thrombin Generation and Fibrinolysis in Vascular Inflammation in Zucker Rats and Evolution with Aging.

The metabolic syndrome (MetS) and aging are associated with modifications in blood coagulation factors, vascular inflammation, and increased risk of thrombosis. Our aim was to determine concomitant changes in thrombin generation in the blood compartment and at the surface of vascular smooth muscle cells (VSMCs) and its interplay with adipokines, free fatty acids (FFA), and metalloproteinases (MMPs) in obese Zucker rats that share features of the human MetS. Obese and age-matched lean Zucker rats were compared at 25 and 80 weeks of age. Thrombin generation was assessed by calibrated automated thrombography (CAT). Endogenous thrombin potential (ETP) was increased in obese rats independent of platelets and age. Clot half-lysis time was delayed with and age. Interleukin (IL)-1β and IL-13 were increased with and age respectively. Addition of exogenous fibrinogen, leptin, linoleic, or increased thrombin generation in plasma whereas adiponectin had an opposite effect. ETP was increased at the surface of VSMCs from obese rats and addition of exogenous further enhanced ETP values. Gelatinase activity was increased in aorta at both ages in obese rats and MMP-2 activity was increased in VSMCs from obese rats. Our study demonstrated in MetS an early prothrombotic phenotype of the blood compartment reinforced by procoagulant properties of dedifferentiated and inflammatory VSMCs. Mechanisms involved (1) increased fibrinogen and impaired fibrinolysis and (2) increased saturated fatty acids responsible for additive procoagulant effects. Whether specifically targeting this hypercoagulability using direct thrombin inhibitors would improve outcome in MetS is worth investigating.

Keyword: obesity

Early and reversible changes to the hippocampal proteome in mice on a high-fat diet.

The rise in global makes it crucial to understand how diet drives -related health conditions, such as premature cognitive decline and Alzheimer\'s disease (AD). In AD hippocampal-dependent episodic memory is one of the first types of memory to be impaired. Previous studies have shown that in mice fed a high-fat diet (HFD) episodic memory is rapidly but reversibly impaired.In this study we use hippocampal proteomics to investigate the effects of HFD in the hippocampus. Mice were fed either a low-fat diet (LFD) or HFD containing either 10% or 60% (Kcal) from fat for 3\u2009days, 1\u2009week or 2\u2009weeks. One group of mice were fed the HFD for 1\u2009week and then returned to the LFD for a further week. Primary hippocampal cultures were challenged with (PA), the most common long-chain saturated FA in the Western diet, and with the anti-inflammatory, n-3 polyunsaturated FA, docosahexaenoic (DHA), or a combination of the two to ascertain effects of these fatty acids on dendritic structure.HFD-induced changes occur in hippocampal proteins involved in metabolism, inflammation, cell stress, cell signalling, and the cytoskeleton after 3\u2009days, 1\u2009week and 2\u2009weeks of HFD. Replacement of the HFD after 1\u2009week by a low-fat diet (LFD) for a further week resulted in partial recovery of the hippocampal proteome. Microtubule-associated protein 2 (MAP2), one of the earliest proteins changed, was used to investigate the impact of fatty acids (FAs) on hippocampal neuronal morphology. PA challenge resulted in shorter and less arborised dendrites while DHA had no effect when applied alone but counteracted the effects of PA when FAs were used in combination. Dendritic morphology recovered when PA was removed from the cell culture media.This study provides evidence for the rapid and reversible effects of diet on the hippocampal proteome and the impact of PA and DHA on dendritic structure.

Keyword: obesity

MiR27a Promotes the Development of Macrophage-like Characteristics in 3T3-L1 Preadipocytes.

Recruitment and polarization of classically activated (M1) macrophages within adipose tissue contribute to chronic low-grade inflammation in . Adipose tissue precursor cells exhibit the capacity to develop macrophage-like characteristics and adipocyte-derived miR27a is known to promote reprogramming of somatic cells. It was unknown whether exogenous addition of miR27a promote the development of macrophage-like characteristics of adipose precursor cells. We examined macrophage surface antigen, phagocytosis and migration ability in 3T3-L1 preadipocytes transfected with miR27a mimics. Transfection of 3T3-L1 preadipocytes with miR27a mimics increased phagocytosis and migration and increased the number of cells expressing the macrophage makers F4/80 and MHC compared to controls. M2 and CD206 macrophage markers were unaltered. In addition, transfection of 3T3-L1 preadipocytes with miR27a mimics reduced PPARγ expression, activated NF-κB and promoted secretion of the inflammatory cytokines MCP-1, TNF-α and IL-1β compared to controls. The level of anti-inflammatory factors Arg-1, IL-10, Ym1 and Fizz1 were unaltered. Secretion of miR27a was increased in conditioned medium prepared from -treated differentiated 3T3-L1 adipocytes compared to controls. Incubation of 3T3-L1 preadipocytes with this conditioned medium increased phagocytosis and migration compared to controls. Finally, conditioned medium prepared from differentiated 3T3-L1 adipocytes transfection with miR27a inhibitors reduced phagocytosis and migration in 3T3-L1 preadipocytes compared to controls. The data indicate that PPARγ agonists may reverse the activation of NF-κB pathway mediated by miR27a overexpression and reduce phagocytosis and migration of adipose precursor cells. In addition, miR27a may promote the development of macrophage-like characteristics in 3T3-L1 preadipocytes.

Keyword: obesity

Analysis of Fatty Esters of Hydroxyl Fatty in Selected Plant Food.

Metabolic syndrome, characterized by , low-grade inflammation, insulin resistance, hyperglycemia, dyslipidemia and hypertension, is a major risk factor for cardiovascular mortality. Preclinical studies on recently discovered classes of lipids - fatty esters of hydroxy fatty acids (FAHFA) have revealed their anti-inflammatory and insulin-sensitizing potential. The FAHFA levels are significantly decreased in insulin-resistant individuals, their application exhibited anti-inflammatory effects and restoring the glucose-insulin homeostasis. The aim of our research was to analyze the overall FAHFA composition in a common diet, as only a partial FAHFA composition has been revealed so far (only the PAHSA subclass was analyzed in a few foods). A new approach to the FAHFAs analysis includes nano-LC and post-column modifier followed by negative ion mass spectrometry, in order to obtain maximum sensitivity. Analysis of different foods - oat (whole grain, coarse flakes and fine flakes), apple, clementine, lemon, strawberry, blueberry, mango, kiwi, avocado, pineapple, banana, onion, garlic, cherry tomato, carrot, parsley root, pepper and radish - exhibited wide inter-food variation in the FAHFA profiles. Sixteen analyzed FAHFAs (, oleic, palmitoleic and stearic hydroxy-esters) showed microgram to low nanogram levels (0.165\xa0ng/g - 32\xa0μg/g FW), with the highest abundancy in oat, clementine, garlic and pineapple. Stearic hydroxy stearic (SAHSA) was the most abundant FAHFA, especially in the food with antioxidative, anti-inflammatory and beneficial metabolic effects. In contrary, the PAHSA - previously proven to have the strongest antihyperglycemic and insulin-sensitizing effects, was not present in some foods (radish, avocado, mango, lemon, cherry tomato, kiwi). Our study proves the importance of overall FAHFA analysis in food (especially in a functional food), because of their potential metabolic benefits and possible future incorporation in special diets.

Keyword: obesity

Effects of saturated and omega-3 polyunsaturated fatty acids on Sertoli cell apoptosis.

is believed to negatively affect male semen quality and is accompanied by dysregulation of free fatty (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive oxygen species were elevated. Furthermore, we demonstrated by in vitro assays that saturated (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated fatty acids might be the cause of -induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs.FFA: free fatty ; HFD: high-fat diet; SD: standard diet; PA: ; PUFA: polyunsaturated fatty ; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive oxygen species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic fatty liver disease; DCFH-DA: 2\', 7\' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic ; PI: propidium iodide; DHA: docosahexenoic .

Keyword: obesity

Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy.

-related non-alcoholic fatty liver disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high-fat diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of fatty liver disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: obesity

Dietary habits affect fatty composition of visceral adipose tissue in subjects with colorectal cancer or .

Aim of this study was to identify a possible relationship among dietary fatty acids (FA) intake, FA adipose tissue (AT) profile and cancer condition in lean vs obese subjects affected or not by colorectal cancer (CRC). Actually, inadequate dietary habits together with physical inactivity are primary determinants of and cancer risk. Changes in lipid metabolism play a crucial role in different types of cancer and key enzymes involved in lipid-metabolic pathways, such as stearoyl-coA-desaturase 1 (SCD-1), are differentially expressed in normal and cancer tissues.Food frequency questionnaires (FFQ) were analyzed by Winfood software. FA were assessed by gas-liquid chromatography in visceral AT samples. Estimated desaturase activities were calculated as precursor FA/product FA ratio. Desaturase gene expressions were evaluated by RT-qPCR.Lean and obese CRC subjects showed inadequate dietary habits. In particular, lean CRC subjects showed increase in the intake of saturated FA, specifically (p\u2009=\u20090.0042) and stearic (p\u2009=\u20090.0091), and a corresponding reduction of monounsaturated FA consumption, in particular oleic (p\u2009=\u20090.002) with respect to lean without CRC. Estimated SCD-1 activity in AT was increased in all the groups vs lean without CRC (pANOVA\u2009=\u20090.029).Unhealthy eating habits, characterizing obese and CRC subjects, may influence the visceral AT profile and contribute to the alteration of the metabolic pathways. The quality of the diet, other than the quantity, can have a main role in the establishment of inflammatory microenvironment and in metabolic changes favouring CRC.

Keyword: obesity

9-PAHSA promotes browning of white fat via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white fat. 9-PAHSA, an endogenous mammalian lipid, which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown fat specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown fat markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of .Copyright © 2018. Published by Elsevier Inc.

Keyword: obesity

Linoleic rescues microglia inflammation triggered by saturated fatty .

Elevated saturated free fatty levels during over-nutrition lead to hypothalamic inflammation, which perturbs energy homeostasis. Whether brain-derived metabolites are coupled to the development of pathogenesis during the early over-nutrition period has not been thoroughly investigated. In this study, we found increased linoleic , an unsaturated fatty , in both the whole brain and hypothalamus of mice fed a high-fat diet for 4 weeks. Furthermore, we observed that linoleic effectively reversed the inflammatory responses induced by treatment in microglial cells. Collectively, this study suggests the reversible function of linoleic on brain inflammation in association with microglial activation during short-term exposure to a high-fat diet.Copyright © 2019. Published by Elsevier Inc.

Keyword: obesity

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance.

Patients with type 2 diabetes (T2D) and/or insulin resistance (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the insulin resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased insulin-stimulated glucose and FA uptake, as well as loss of endosomal acidification and insulin signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to , diabetes and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: obesity

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut microbiota and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: obesity

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: obesity

Telmisartan protects against high glucose/high lipid-induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: obesity

CTRP3 attenuates high-fat diet-induced male reproductive dysfunction in mice.

Recent studies have suggested a role for abdominal in male infertility. Previous studies have found that cell apoptosis exerts an important role in -related male infertility. C1q/TNF-related protein 3 (CTRP3), a paralog of adiponectin, has been proposed to exert anti-apoptotic effects and to attenuate diabetes-related cardiac injuries. However, the role of CTRP3 in high-fat diet (HFD)-induced spermatogenic impairment remains unclear. In the present study, we fed male mice an HFD for 24 weeks to induce . The expression of CTRP3 was decreased by HFD feeding. Supplementation with the recombinant human globular domain of CTRP3 (0.25 μg/g/day) for 4 weeks beginning at 20 weeks of the HFD improved spermatogenic function in the HFD-fed mice, which were characterized by improved testis morphology, increased testis weight/body weight ratio, and increased sperm count, sperm viability, and sperm motility. We also found that CTRP3 infusion resulted in the attenuation of endoplasmic reticulum (ER) stress and the activation of silence information regulator 1 (SIRT1) in the testes of obese mice. Our study also suggested that CTRP3 attenuated the (PA)-induced reductions in sperm viability and motility via the inhibition of ER stress. Moreover, germ cell-specific knockout abolished the protective effects of CTRP3 and studies of human sperm showed that the protective effects of CTRP3 on sperm viability and motility were abrogated by a specific inhibitor of SIRT1. Thus, our results demonstrated that CTRP3 expression protected against HFD-induced spermatogenic deficiency through the SIRT1/ER stress pathway.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: obesity

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and . We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: obesity

MicroRNA-194 inhibition improves dietary-induced non-alcoholic fatty liver disease in mice through targeting on FXR.

Non-alcoholic fatty liver disease (NAFLD) affects -associated metabolic syndrome, which exhibits hepatic steatosis, insulin insensitivity and glucose intolerance. Previous studies indicated that hepatic microRNAs (miRs) play critical roles in the development of NAFLD. In this study, we aim to explore the pathophysiological role of miR-194 in -mediated metabolic dysfunction. Our findings show that the high fat diet or treatment significantly increase hepatic miR-194 levels in vivo and in vitro. Silence of miR-194 protects -induced inflammatory response in cultured hepatocytes, and attenuates structural disorders, lipid deposits and inflammatory response in fatty liver. MiR-194 inhibitor also improves glucose and insulin intolerance in obese mice. Through dual luciferase assay, we demonstrate that miR-194 directly binds to FXR/Nr1h4 3\'-UTR, and inhibits gene expression of FXR/Nr1h4. Furthermore, overexpression of miR-194 downregulates FXR/Nr1h4 in cultured hepatocytes, but miR-194 inhibitor reversely increases FXR/Nr1h4 expression in obese mouse liver tissues. On the contrast, silence of FXR/Nr1h4 abolishes the hepatic benefits in obese mice treated with miR-194 inhibitor. Present study provides a novel finding that suppression of miR-194 attenuates dietary-induced NAFLD via upregulation of FXR/Nr1h4. The findings suggest miR-194/FXR are potential diagnostic markers and therapeutic targets for NAFLD.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: obesity

induces guanylin gene expression through the toll-like receptor 4/nuclear factor κB pathway in rat macrophages.

Recently, we showed that double-transgenic rats overexpressing guanylin (Gn), a bioactive peptide, and its receptor, guanylyl cyclase-C (GC-C), specifically in macrophages demonstrate an anti- phenotype and low expression levels of proinflammatory cytokines in the mesenteric fat even when fed a high-fat diet.Here, we examined the levels and mechanism of Gn and GC-C transcription following saturated fatty and lipopolysaccharide (LPS), an activator of toll-like receptor 4 (TLR4), exposure by using the NR8383 macrophage cell line. In addition, the levels of guanylin and cGMP were increased by addition of either or LPS.Next, we investigated the interaction of the gene transcription and nuclear factor κB (NF-κB) by using an NF-κB inhibitor and chromatin immunoprecipitation assay. We showed that induced Gn gene expression via TLR4 and NF-κB. Moreover, we demonstrated that NF-κB binding to the Gn promoter was responsible for the induction of gene transcription by or LPS.Our results indicate that saturated fatty acids such as acidactivate Gn gene expression via the NF-κB pathway, raising the possibility that the activated Gn-GC-C system may contribute to the inhibition of high-fat diet-induced proinflammatory cytokines in macrophages.

Keyword: obesity

Downregulation of MALAT1 alleviates saturated fatty -induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis.

The increased level of saturated fatty acids (SFAs) is found in patients with diabetes, , and other metabolic disorders. SFAs can induce lipotoxic damage to cardiomyocytes, but the mechanism is unclear. The long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acts as a key regulator in (PA)-induced hepatic steatosis, but its role in PA-induced myocardial lipotoxic injury is still unknown. The aim of this study was to explore the role and underlying mechanism of MALAT1 in PA-induced myocardial lipotoxic injury. MALAT1 expression in PA-treated human cardiomyocytes (AC16 cells) was detected by RT-qPCR. The effect of MALAT1 on PA-induced myocardial injury was measured by Cell Counting Kit-8, lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) assays. Apoptosis was detected by flow cytometry. The activities of cytokines and nuclear factor (NF)-κB were detected by enzyme-linked immunosorbent assay. The interaction between MALAT1 and miR-26a was evaluated by a luciferase reporter assay and RT-qPCR. The regulatory effects of MALAT1 on high mobility group box 1 (HMGB1) expression were evaluated by RT-qPCR and western blotting. MALAT1 was significantly upregulated in cardiomyocytes after PA treatment. Knockdown of MALAT1 increased the viability of PA-treated cardiomyocytes, decreased apoptosis, and reduced the levels of LDH, CK-MB, TNF-α, and IL-1β. Moreover, we found that MALAT1 specifically binds to miR-26a and observed a reciprocal negative regulatory relationship between these factors. We further found that the downregulation of MALAT1 represses HMGB1 expression, thereby inhibiting the activation of the Toll-like receptor 4 (TLR4)/NF-κB-mediated inflammatory response. These repressive effects were rescued by an miR-26a inhibitor. We demonstrate that MALAT1 is induced by SFAs and its downregulation alleviates SFA-induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis. Our findings provide new insight into the mechanism underlying myocardial lipotoxic injury.

Keyword: obesity

High-density lipoprotein ameliorates -induced lipotoxicity and oxidative dysfunction in H9c2 cardiomyoblast cells via ROS suppression.

High levels circulating saturated fatty acids are associated with diabetes, and hyperlipidemia. In heart, the accumulation of saturated fatty acids has been determined to play a role in the development of heart failure and diabetic cardiomyopathy. High-density lipoprotein (HDL) has been reported to possess key atheroprotective biological properties, including cellular cholesterol efflux capacity, anti-oxidative and anti-inflammatory activities. However, the underlying mechanisms are still largely unknown. Therefore, the aim of the present study is to test whether HDL could protect (PA)-induced cardiomyocyte injury and explore the possible mechanisms.H9c2 cells were pretreated with HDL (50-100\u2009μg/ml) for 2\u2009h followed by PA (0.5\u2009mM) for indicated time period. Our results showed that HDL inhibited PA-induced cell death in a dose-dependent manner. Moreover, HDL rescued PA-induced ROS generation and the phosphorylation of JNK which in turn activated NF-κB-mediated inflammatory proteins expressions. We also found that PA impaired the balance of BCL family proteins, destabilized mitochondrial membrane potential, and triggered subsequent cytochrome c release into the cytosol and activation of caspase 3. These detrimental effects were ameliorated by HDL treatment.PA-induced ROS accumulation and results in cardiomyocyte apoptosis and inflammation. However, HDL attenuated PA-induced lipotoxicity and oxidative dysfunction via ROS suppression. These results may provide insight into a possible molecular mechanism underlying HDL suppression of the free fatty -induced cardiomyocyte apoptosis.

Keyword: obesity

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and lipid metabolism.

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet lipid metabolism in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high-fat diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced lipid accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced lipid deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell lipid accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: obesity

Perivascular adipose tissue dysfunction aggravates adventitial remodeling in obese mini pigs via NLRP3 inflammasome/IL-1 signaling pathway.

Perivascular adipose tissue (PVAT), a special type of adipose tissue, closely surrounds vascular adventitia and produces numerous bioactive substances to maintain vascular homeostasis. PVAT dysfunction has a crucial role in regulating vascular remodeling, but the exact mechanisms remain unclear. In this study, we investigated whether and how -induced PVAT dysfunction affected adventitia remodeling in early vascular injury stages. Mini pigs were fed a high sugar and fat diet for 6 months to induce metabolic syndrome and . In the mini pigs, left carotid vascular injury was then generated using balloon dilation. Compared with normal mini pigs, obese mini pigs displayed significantly enhanced vascular injury-induced adventitial responses, evidenced by adventitia fibroblast (AF) proliferation and differentiation, and adventitia fibrosis, as well as exacerbated PVAT dysfunction characterized by increased accumulation of resident macrophages, particularly the M1 pro-inflammatory phenotype, increased expression of leptin and decreased expression of adiponectin, and production of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Primary AFs cultured in PVAT-conditioned medium from obese mini pigs also showed significantly increased proliferation and differentiation. We further revealed that activated nod-like receptor protein 3 (NLRP3) inflammasome and its downstream products, i.e., IL-1 family members such as IL-1β and IL-18 were upregulated in the PVAT of obese mini pigs; PVAT dysfunction was also demonstrated in preadipocytes treated with . Finally, we showed that pretreatment with IL-1 receptor (IL-1R) antagonist or IL-1R knockdown blocked AF proliferation and differentiation in AFs cultured in PVAT-conditioned medium. These results demonstrate that -induced PVAT dysfunction aggravates adventitial remodeling after early vascular injury with elevated AF proliferation and differentiation via activating the NLRP3/IL-1 signaling pathway.

Keyword: obesity

Role of the saturated fatty palmitate in the interconnected hypothalamic control of energy homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body energy and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, , and type 2 diabetes mellitus. Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and energy homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: obesity

Anthocyanins ameliorate palmitate-induced inflammation and insulin resistance in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade inflammation leading to insulin resistance. Adipocyte differentiation inhibitors are expected to be effective in preventing and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from metabolic stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, inflammation, and insulin resistance in 3T3-L1 adipocytes. ACN extract pretreatment reduces lipid accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces inflammation with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces insulin resistance by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce insulin pathway with levels higher than insulin control cells, supporting an insulin sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against comorbidities, due to their protective effects against inflammation/insulin resistance in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and insulin resistance conditions related to .© 2019 John Wiley & Sons, Ltd.

Keyword: obesity

1,25-Dihydroxyvitamin D regulates lipid metabolism and glucose utilization in differentiated 3T3-L1 adipocytes.

It is well established that adipose tissue can both store and metabolize vitamin D. The active form of vitamin D, 1,25 dihydroxyvitamin D [1,25(OH)D], regulates adipocyte differentiation and inflammation, highlighting the multifaceted role that vitamin D plays in adipose tissue physiology. However, there is limited evidence regarding vitamin D regulation of mature adipocyte lipid metabolism. We hypothesize that 1,25(OH)D alters lipid and glucose metabolism in differentiated 3T3-L1 adipocytes to reduce triacylglycerol (TAG) accumulation. In this study, 1,25(OH)D (10\u202fnmol/L) stimulated a 21% reduction in TAG accumulation in differentiated 3T3-L1 adipocytes after 4\u202fdays (P\u202f=\u202f.01) despite a significant increase in fatty uptake (P\u202f<\u202f.01). Additionally, 1,25(OH)D stimulated a 2.5-fold increase in CO production from [1-C] (P\u202f<\u202f.01), indicative of an elevated rate of fatty β-oxidation, while stimulating a 9% reduction in de novo fatty synthesis (P\u202f=\u202f.03). Interestingly, d-[U-C]glucose incorporation into fatty acids was reduced by 30% in response to 1,25(OH)D (P\u202f<\u202f.01), indicating a reduced contribution of glucose as a substrate for de novo lipogenesis. Consistent with these findings, mRNA expression of the anaplerotic enzyme pyruvate carboxylase was reduced by 41% (P\u202f<\u202f.01). In summary, 1,25(OH)D stimulated fatty oxidation and reduced TAG accumulation in differentiated adipocytes. Furthermore, 1,25(OH)D reduced glucose utilization as a substrate for fatty synthesis potentially by downregulating pyruvate carboxylase and stimulating glucose disposal as glycerol. Collectively, these 1,25(OH)D-induced changes in lipid metabolism and glucose utilization may contribute to the reduction in TAG accumulation and be protective against excessive fat mass accumulation and associated metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Neuroprotective effects of vitamin D on high fat diet- and -induced enteric neuronal loss in mice.

The role of vitamin D in and diabetes is debated. Obese and/or diabetic patients have elevated levels of free fatty acids, increased susceptibility to gastrointestinal symptoms and are suggested to have altered vitamin D balance. The enteric nervous system is pivotal in regulating gastrointestinal activity and high fat diet (HFD) has been shown to cause loss of enteric neurons in ileum and colon. This study investigates the effect of vitamin D on HFD- and -induced enteric neuronal loss in vivo and in vitro.Mice were fed either a normal diet (ND) or HFD supplemented with varying levels of vitamin D (from 0x to 20x normal vitamin D level) for 19 weeks. Ileum and colon were analyzed for neuronal numbers and remodeling. Primary cultures of myenteric neurons from mouse small intestine were treated with (4x10M) and/or 1α,25-hydroxy-vitamin D3 (VD, 10- 10M) with or without modulators of lipid metabolism and VD pathways. Cultures were analyzed by immunocyto- and histochemical methods.Vitamin D supplementation had no effect on enteric neuronal survival in the ND group. HFD caused substantial loss of myenteric neurons in ileum and colon. Vitamin D supplementation between 0-2x normal had no effect on HFD-induced neuronal loss. Supplementation with 20x normal, prevented the HFD-induced neuronal loss. In vitro supplementation of VD prevented the -induced neuronal loss. The VD receptor (VDR) was not identified in enteric neurons. Enteric glia expressed the alternative VD receptor, protein disulphide isomerase family A member 3 (PDIA3), but PDIA3 was not found to mediate the VD response in vitro. Inhibition of peroxisome proliferator-activated receptor gamma (PPARγ) and immune neutralization of isocitrate lyase prevented the VD mediated neuroprotection to exposure.Results show that VD protect enteric neurons against HFD and induced neuronal loss. The mechanism behind is suggested to be through activation of PPARγ leading to improved neuronal peroxisome function and metabolism of neuronal lipid intermediates.

Keyword: obesity

High Fat Diet Upregulates Fatty Oxidation and Ketogenesis via Intervention of PPAR-γ.

Systemic hyperlipidemia and intracellular lipid accumulation induced by chronic high fat diet (HFD) leads to enhanced fatty oxidation (FAO) and ketogenesis. The present study was aimed to determine whether activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) by surplus free fatty acids (FA) in hyperlipidemic condition, has a positive feedback regulation over FAO and ketogenic enzymes controlling lipotoxicity and cardiac apoptosis.8 weeks old C57BL/6 wild type (WT) or PPAR-γ-/- mice were challenged with 16 weeks 60% HFD to induce mediated type 2 diabetes mellitus (T2DM) and diabetic cardiomyopathy. Treatment course was followed by echocardiographic measurements, glycemic and lipid profiling, immunoblot, qPCR and immunohistochemistry (IHC) analysis of PPAR-γ and following mitochondrial metabolic enzymes 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), mitochondrial β- hydroxy butyrate dehydrogenase (BDH1) and pyruvate dehydrogenase kinase isoform 4 (PDK4). In vivo model was translated in vitro, with neonatal rat cardiomyocytes (NRCM) treated with PPAR-γ agonist/antagonist and PPAR-γ overexpression adenovirus in presence of (PA). Apoptosis was determined in vivo from left ventricular heart by TUNEL assay and immunoblot analysis.We found exaggerated circulating ketone bodies production and expressions of the related mitochondrial enzymes HMGCS2, BDH1 and PDK4 in HFD-induced diabetic hearts and in PA-treated NRCM. As a mechanistic approach we found HFD mediated activation of PPAR-γ is associated with the above-mentioned mitochondrial enzymes. HFD-fed PPAR-γ-/-mice display decreased hyperglycemia, hyperlipidemia associated with increased insulin responsiveness as compared to HFD-fed WT mice PPAR-γ-/-HFD mice demonstrated a more robust functional recovery after diabetes induction, as well as significantly reduced myocyte apoptosis and improved cardiac function.PPAR-γ has been described previously to regulate lipid metabolism and adipogenesis. The present study suggests for the first time that increased PPAR-γ expression by HFD is responsible for cardiac dysfunction via upregulation of mitochondrial enzymes HMGCS2, BDH1 and PDK4. Targeting PPAR-γ and its downstream mitochondrial enzymes will provide novel strategies in preventing metabolic and myocardial dysfunction in diabetes mellitus.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: obesity

Chokeberry Juice Containing Polyphenols Does Not Affect Cholesterol or Blood Pressure but Modifies the Composition of Plasma Phospholipids Fatty Acids in Individuals at Cardiovascular Risk.

Chokeberry polyphenols have been suggested to reduce cholesterol and blood pressure and thus protect against cardiovascular diseases (CVD), but the evidence in humans is limited and inconsistent. This randomized double-blinded three-parallel groups trial investigated the changes in various anthropometric and clinical biomarkers, and in plasma phospholipids fatty acids (PPFA) in volunteers at cardiovascular risk after a four-week intervention with 100 mL/day of (1) chokeberry juice with a high-dose of polyphenols (1177.11 mg gallic equivalents, GAE); (2) chokeberry juice with a low-dose of polyphenols (294.28 mg GAE) and; (3) a nutritionally matched polyphenol-free placebo drink. Our results indicate that the intake of chokeberry juice containing either the low or the high dose of polyphenols cannot be linked with a reduction in total- and low-density lipoprotein (LDL)cholesterol or in systolic (SBP) and diastolic (DBP) blood pressure in comparison with the consumption of the placebo drink. However, we found evidence of moderate changes in the PPFA, i.e., increased saturated fatty acids (SFA), mostly , and reduced n-6 polyunsaturated fatty acids (PUFA), principally linoleic (LA) with the intake of chokeberry against the placebo. These effects may be associated with the polyphenols but we could not differentiate a clear dose-response effect. Further research is still needed to elucidate the contribution of the polyphenolic fraction to the potential cardiovascular effects of the chokeberry and to build up the evidence of its potential benefit via the modulation of PPFA composition.

Keyword: obesity

The traditional uses, phytochemistry, and pharmacology of Atractylodes macrocephala Koidz.: A review.

Atractylodes macrocephala Koidz. (called Baizhu in China) is a medicinal plant that has long been used as a tonic agent in various ethno-medical systems in East Asia, especially in China, for the treatment of gastrointestinal dysfunction, cancer, osteoporosis, , and fetal irritability.This review aims to provide a systematic summary on the botany, traditional uses, phytochemistry, pharmacology, pharmacokinetics, and toxicology of A. macrocephala to explore the future therapeutic potential and scientific potential of this plant.A literature search was performed on A. macrocephala using scientific databases including Web of Science, Google Scholar, Baidu Scholar, Springer, PubMed, SciFinder, and ScienceDirect. Information was also collected from classic books of Chinese herbal medicine, Ph.D. and M.Sc. dissertations, unpublished materials, and local conference papers on toxicology. Plant taxonomy was confirmed to the database "The Plant List" (www.theplantlist.org).More than 79 chemical compounds have been isolated from A. macrocephala, including sesquiterpenoids, triterpenoids, polyacetylenes, coumarins, phenylpropanoids, flavonoids and flavonoid glycosides, steroids, benzoquinones, and polysaccharides. Crude extracts and pure compounds of A. macrocephala are used to treat gastrointestinal hypofunction, cancer, arthritis, osteoporosis, splenic asthenia, abnormal fetal movement, Alzheimer disease, and . These extracts have various pharmacological effects, including anti-tumor activity, anti-inflammatory activity, anti-aging activity, anti-oxidative activity, anti-osteoporotic activity, neuroprotective activity, and immunomodulatory activity, as well as improving gastrointestinal function and gonadal hormone regulation.A. macrocephala is a valuable traditional Chinese medicinal herb with multiple pharmacological activities. Pharmacological investigations support the traditional use of A. macrocephala, and may validate the folk medicinal use of A. macrocephala to treat many chronic diseases. The available literature shows that much of the activity of A. macrocephala can be attributed to sesquiterpenoids, polysaccharides and polyacetylenes. However, there is a need to further understand the molecular mechanisms and the structure-function relationship of these constituents, as well as their potential synergistic and antagonistic effects. Further research on the comprehensive evaluation of medicinal quality, the understanding of multi-target network pharmacology of A. macrocephala, as well as its long-term in vivo toxicity and clinical efficacy is recommended.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

Inhibition of hepatocyte nuclear factor 1b induces hepatic steatosis through DPP4/NOX1-mediated regulation of superoxide.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disorder that is closely associated with insulin resistance and type 2 diabetes. Previous studies have suggested that hepatocyte nuclear factor 1b (HNF1b) ameliorates insulin resistance. However, the role of HNF1b in the regulation of lipid metabolism and hepatic steatosis remains poorly understood. We found that HNF1b expression was decreased in steatotic livers. We injected mice with lentivirus (LV) expressing HNF1b shRNA to generate mice with hepatic knockdown of HNF1b. We also injected high fat (HF) diet-induced obese and db/db diabetic mice with LV expressing HNF1b to overexpress HNF1b. Knockdown of HNF1b increased hepatic lipid contents and induced insulin resistance in mice and in hepatocytes. Knockdown of HNF1b worsened HF diet-induced increases in hepatic lipid contents, liver injury and insulin resistance in mice and PA-induced lipid accumulation and impaired insulin signaling in hepatocytes. Moreover, overexpression of HNF1b alleviated HF diet-induced increases in hepatic lipid content and insulin resistance in mice. Knockdown of HNF1b increased expression of genes associated with lipogenensis and endoplasmic reticulum (ER) stress. DPP4 and NOX1 expression was increased by knockdown of HNF1b and HNF1b directly bound with the promoters of DPP4 and NOX1. Overexpression of DPP4 or NOX1 was associated with an increase in lipid droplets in hepatocytes and decreased expression of DPP4 or NOX1 suppressed the effects of knockdown of HNF1b knockdown on triglyceride (TG) formation and insulin signaling. Knockdown of HNF1b increased superoxide level and decreased glutathione content, which was inhibited by downregulation of DPP4 and NOX1. N-acetylcysteine (NAC) suppressed HNF1b knockdown-induced ER stress, TG formation and insulin resistance. (PA) decreased HNF1b expression which was inhibited by NAC. Taken together, these studies demonstrate that HNF1b plays an essential role in controlling hepatic TG homeostasis and insulin sensitivity by regulating DPP4/NOX1mediated generation of superoxide.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: obesity

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for and diabetes.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: obesity

Celastrol reverses (PA)-caused TLR4-MD2 activation-dependent insulin resistance via disrupting MD2-related cellular binding to PA.

Elevated plasma statured fatty acids (FFAs) cause TLR4/MD2 activation-dependent inflammation and insulin tolerance, which account for the occurrence and development of . It has been confirmed that statured (PA) (the most abundant FFA) could bind MD2 to cause cellular inflammation. The natural compound celastrol could improve , which is suggested via inhibiting inflammation, yet the detailed mechanism for celastrol is still unclear. As celastrol is reported to directly target MD2, we thought disrupting the binding between FFAs and MD2 might be one of the ways for celastrol to inhibit FFAs-caused inflammation and insulin resistance. In this study, we found evidence to support our hypothesis: celastrol could reverse PA-caused TLR4/MD2 activation-dependent insulin resistance, as determined by glucose-lowering ability, cellular glucose uptake, insulin action-related proteins and TLR4/MD2/NF-κB activation. Bioinformatics and cellular experiments showed that both celastrol and PA could bind MD2, and that celastrol could expel PA from cells. Finally, celastrol could reverse high fat diet caused hyperglycemia and , and liver NF-kB activations. Taking together, we proved that celastrol could reverses PA-caused TLR4-MD2 activation-dependent insulin resistance via disrupting PA binding to MD2.© 2018 Wiley Periodicals, Inc.

Keyword: obesity

The autophagic degradation of Cav-1 contributes to PA-induced apoptosis and inflammation of astrocytes.

The accumulation of (PA), implicated in , can induce apoptotic cell death and inflammation of astrocytes. Caveolin-1 (Cav-1), an essential protein for astrocytes survival, can be degraded by autophagy, which is a double-edge sword that can either promote cell survival or cell death. The aim of this study was to delineate whether the autophagic degradation of Cav-1 is involved in PA-induced apoptosis and inflammation in hippocampal astrocytes. In this study we found that: (1) PA caused apoptotic death and inflammation by autophagic induction; (2) Cav-1 was degraded by PA-induced autophagy and PA induced autophagy in a Cav-1-independent manner; (3) the degradation of Cav-1 was responsible for PA-induced autophagy-dependent apoptotic cell death and inflammation; (4) chronic high-fat diet (HFD) induced Cav-1 degradation, apoptosis, autophagy, and inflammation in the hippocampal astrocytes of rats. Our results suggest that the autophagic degradation of Cav-1 contributes to PA-induced apoptosis and inflammation of astrocytes. Therefore, Cav-1 may be a potential therapeutic target for central nervous system injuries caused by PA accumulation.

Keyword: obesity

and Oleic : The Yin and Yang of Fatty Acids in Type 2 Diabetes Mellitus.

Increased plasma non-esterified fatty acids (NEFAs) link with insulin resistance and type 2 diabetes mellitus (T2DM). However, in contrast to the saturated FA (SFA) , the monounsaturated FA (MUFA) oleic elicits beneficial effects on insulin sensitivity, and the dietary :oleic ratio impacts diabetes risk in humans. Here we review recent mechanistic insights into the beneficial effects of oleic compared with on insulin resistance and T2DM, including its anti-inflammatory actions, and its capacity to inhibit endoplasmic reticulum (ER) stress, prevent attenuation of the insulin signaling pathway, and improve β cell survival. Understanding the molecular mechanisms of the antidiabetic effects of oleic may contribute to understanding the benefits of this FA in the prevention or delay of T2DM.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: obesity

Cardiomyocyte-specific knockout of endothelin receptor a attenuates cardiomyopathy.

Endothelin (ET)-1 is implicated in the pathophysiology of cardiovascular diseases although its role in anomalies has not been fully elucidated. This study was designed to examine the impact of ET-1 receptor A (ET) ablation on -induced changes in cardiac geometry and contractile function, as well as the mechanisms involved with a focus on autophagy. Cardiomyocyte-specific ET receptor knockout (ETAKO) and WT mice were fed either low-fat (10% calorie from fat) or high-fat (45% calorie from fat) diet for 24\u202fweeks. Glucose tolerance test was examined to confirm insulin resistance. High-fat diet intake compromised myocardial geometry (enlarged left ventricular diameters in systole and diastole), morphology (cardiac hypertrophy, increased wall thickness and interstitial fibrosis), contractile function (reduced fractional shortening, ejection fraction and cardiomyocyte shortening) and intracellular Ca handling, the effect of which was significantly attenuated by ETAKO. TUNEL staining revealed overt apoptosis in high-fat-fed group, the effect was reverted by ETAKO. Western blot analysis noted that high-fat intake downregulated leptin receptor and PPARγ, insulin signaling (elevated basal/dampened insulin-stimulated phosphorylation of Akt and IRS1), phosphorylation of AMPK, ACC, upregulated GATA-4, ANP, NFATc3, PPARα, m-TOR/p70s6k signaling, which were attenuated by ETAKO with the exception of AMPK/ACC. Furthermore, high-fat intake suppressed cardiac autophagy, which was abrogated by ETAKO. In cultured murine cardiomyocytes, challenged mimicked high-fat diet-induced hypertrophic and autophagic responses, the effect of which were abolished by the ET receptor antagonist BQ123 or mTOR inhibitor rapamycin. These results suggest that inhibition of ET rescues high-fat intake-induced cardiac anomalies possibly through autophagy regulation.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

Acne vulgaris: The metabolic syndrome of the pilosebaceous follicle.

Acne vulgaris is an epidemic inflammatory disease of the human sebaceous follicle and represents the most common skin disease affecting about 85% of adolescents in Westernized populations. Acne vulgaris is primarily a disease of wealthy countries and exhibits higher prevalence rates in developed compared with developing countries. No acne has been found in non-Westernized populations still living under Paleolithic dietary conditions constraining hyperglycemic carbohydrates, milk, and dairy products. The high prevalence rates of adolescent acne cannot be explained by the predominance of genetic factors but by the influence of a Western diet that overstimulates the key conductor of metabolism, the nutrient- and growth factor-sensitive kinase mTORC1. Increased mTORC1 activity has been detected in lesional skin and sebaceous glands of acne patients compared with acne-free controls. Increased mTORC1 signaling is a characteristic feature of insulin resistance, , type 2 diabetes mellitus, cancer, and neurodegenerative diseases. Acne vulgaris is a family member of mTORC1-driven diseases of civilization and represents the MetS of the sebaceous follicle.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: obesity

Individual fatty acids in erythrocyte membranes are associated with several features of the metabolic syndrome in obese children.

leads to the clustering of cardiovascular (CV) risk factors and the metabolic syndrome (MetS) also in children and is often accompanied by non-alcoholic fatty liver disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 fatty acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting insulin (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting insulin (r\u2009=\u20090.287). Fatty Liver Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

Keyword: obesity

Ginsenoside Rb1 improves leptin sensitivity in the prefrontal cortex in obese mice.

impairs leptin-induced regulation of brain-derived neurotrophic factor (BDNF) expression and synaptogenesis, which has been considered to be associated with the incidence of neuronal degenerative diseases, cognitive decline, and depression. Ginsenoside Rb1 (Rb1), a major bioactive component of ginseng, is known to have an antiobesity effect and improve cognition. This study examined whether Rb1 can improve central leptin effects on BDNF expression and synaptogenesis in the prefrontal cortex during using an in vivo and an in vitro model.Ginsenoside Rb1 (Rb1) chronic treatment improved central leptin sensitivity, leptin-JAK2-STAT3 signaling, and leptin-induced regulation of BDNF expression in the prefrontal cortex of high-fat diet-induced obese mice. In cultured prefrontal cortical neurons, , the saturated fat, impaired leptin-induced BDNF expression, reduced the immunoreactivity and mRNA expression of synaptic proteins, and impaired leptin-induced neurite outgrowth and synaptogenesis. Importantly, Rb1 significantly prevented these pernicious effects induced by .These results indicate that Rb1 reverses central leptin resistance and improves leptin-BDNF-neurite outgrowth and synaptogenesis in the prefrontal cortical neurons. Thus, Rb1 supplementation may be a beneficial avenue to treat -associated neurodegenerative disorders.© 2017 John Wiley & Sons Ltd.

Keyword: obesity

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

triggers inflammatory responses in N42 cultured hypothalamic cells partially via ceramide synthesis but not via TLR4.

A high-fat diet induces hypothalamic inflammation in rodents which, in turn, contributes to the development of by eliciting both insulin and leptin resistance. However, the mechanism by which long-chain saturated fatty acids trigger inflammation is still contentious. To elucidate this mechanism, the effect of fatty acids on the expression of the pro-inflammatory cytokines IL-6 and TNFα was investigated in the mHypoE-N42 hypothalamic cell line (N42). N42 cells were treated with lauric (LA) and (PA). PA challenge was carried out in the presence of either a TLR4 inhibitor, a ceramide synthesis inhibitor (L-cycloserine), oleic (OA) or eicosapentaenoic (EPA). Intracellular ceramide accumulation was quantified using LC-ESI-MS/MS. PA but not LA upregulated IL-6 and TNFα. L-cycloserine, OA and EPA all counteracted PA-induced intracellular ceramide accumulation leading to a downregulation of IL-6 and TNFα. However, a TLR4 inhibitor failed to inhibit PA-induced upregulation of pro-inflammatory cytokines. In conclusion, PA induced the expression of IL-6 and TNFα in N42 neuronal cells independently of TLR4 but, partially, via ceramide synthesis with OA and EPA being anti-inflammatory by decreasing PA-induced intracellular ceramide build-up. Thus, ceramide accumulation represents one on the mechanisms by which PA induces inflammation in neurons.

Keyword: obesity

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) , 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: obesity

Palmitate aggravates proteinuria-induced cell death and inflammation via CD36-inflammasome axis in the proximal tubular cells of obese mice.

High levels of serum free fatty acids (FFAs) and proteinuria have been implicated in the pathogenesis of -related nephropathy. CD36, a class B scavenger receptor, is highly expressed in the renal proximal tubules and mediates FFA uptake. It is not clear whether FFA- and proteinuria-mediated CD36 activation coordinates NLRP3 inflammasomes to induce renal tubular injury and inflammation. In this study, we investigated the roles of CD36 and NLRP3 inflammasomes in FFA-induced renal injury in high-fat diet (HFD)-induced . HFD-fed C57BL/6 mice and palmitate-treated HK2 renal tubular cells were used as in vivo and in vitro models. Immunohistochemical staining showed that CD36, IL-1β, and IL-18 levels increased progressively in the kidneys of HFD-fed mice. Sulfo- N-succinimidyl oleate (SSO), a CD36 inhibitor, attenuated the HFD-induced upregulation of NLRP3, IL-1β, and IL-18 and suppressed the colocalization of NLRP3 and ASC in renal tubular cells. In vitro, SSO abolished the palmitate-induced activation of IL-1β, IL-18, and caspase-1 in HK2 proximal tubular cells. Furthermore, treatment with SSO and the knockdown of caspase-1 expression by siRNA both inhibited palmitate-induced cell death and apoptosis in HK2 cells. Collectively, palmitate causes renal tubular inflammation, cell death, and apoptosis via the CD36/NLRP3/caspase-1 axis, which may explain, at least in part, the mechanism underlying FFA-related renal tubular injury. The blockade of CD36-induced cellular processes is therefore a promising strategy for treating -related nephropathy.

Keyword: obesity

Downregulation of miR-320 Alleviates Endoplasmic Reticulum Stress and Inflammatory Response in 3T3-L1 Adipocytes.

MicroRNAs serve important roles in the regulation of endoplasmic reticulum stress (ERs). This study aimed to investigate the role of microRNA-320 (miR-320) in the development of ERs and the inflammatory response in 3T3-L1 adipocytes.The adipose tissue expression levels of miR-320 and ERs markers (GRP78, GRP94, Derlin-1 and CHOP) and the serum concentration of inflammatory cytokines (TNF-α, NF-κB and IL-6) in obese patients were evaluated using quantitative real-time RT-PCR. The correlation of miR-320 with genes involved in ERs and inflammation was analyzed. The effects of miR-320 on ERs and inflammation were explored using mature 3T3-L1 adipocytes, which were pretreated with (PA).ERs markers and inflammatory cytokines were all upregulated in obese patients. Adipose tissue miR-320 expression was also increased in obese patients, and had positive correlations with the levels of ERs markers and inflammatory cytokines. After PA treatment, the levels of ERs markers and inflammatory cytokines were elevated significantly in 3T3-L1 adipocytes. Moreover, miR-320 expression was increased in the cells under ERs status. The upregulation of miR-320 could enhance the expression of ERs markers and inflammatory cytokines, but the downregulation of miR-320 resulted in the opposite results.The data of this study indicate that miR-320 expression is upregulated in ERs status, and the downregulation of miR-320 ameliorates ERs and the inflammatory response in 3T3-L1 adipocytes. We consider that the approaches to decrease miR-320 expression may be novel therapeutic strategies for the treatment of and -related diseases.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: obesity

Sphingolipid changes do not underlie fatty -evoked GLUT4 insulin resistance nor inflammation signals in muscle cells.

Ceramides contribute to -linked insulin resistance and inflammation in vivo, but whether this is a cell-autonomous phenomenon is debated, particularly in muscle, which dictates whole-body glucose uptake. We comprehensively analyzed lipid species produced in response to fatty acids and examined the consequence to insulin resistance and pro-inflammatory pathways. L6 myotubes were incubated with BSA-adsorbed palmitate or palmitoleate in the presence of myriocin, fenretinide, or fumonisin B1. Lipid species were determined by lipidomic analysis. Insulin sensitivity was scored by Akt phosphorylation and glucose transporter 4 (GLUT4) translocation, while pro-inflammatory indices were estimated by IκBα degradation and cytokine expression. Palmitate, but not palmitoleate, had mild effects on Akt phosphorylation but significantly inhibited insulin-stimulated GLUT4 translocation and increased expression of pro-inflammatory cytokines and Ceramides, hexosylceramides, and sphingosine-1-phosphate significantly heightened by palmitate correlated negatively with insulin sensitivity and positively with pro-inflammatory indices. Inhibition of sphingolipid pathways led to marked changes in cellular lipids, but did not prevent palmitate-induced impairment of insulin-stimulated GLUT4 translocation, suggesting that palmitate-induced accumulation of deleterious lipids and insulin resistance are correlated but independent events in myotubes. We propose that muscle cell-endogenous ceramide production does not evoke insulin resistance and that deleterious effects of ceramides in vivo may arise through ancillary cell communication.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: obesity

Postprandial Lipid Response to High-Saturated and High-Monounsaturated Fat Meals in Normal-Weight or Overweight Women.

We evaluated postprandial response of the lipid metabolism markers after the intake of a high-saturated fat (HSM) or high-monounsaturated fat meal (HMM).A randomized, controlled and acute intervention study included 63 women (age 26.9 ± 6.1 years): 35 normal weight (NW) and 28 overweight (OW) (total body fat [TBF] 24.7 ± 3.9% and 36.6 ± 3.9%, respectively). After 12\xa0hours of fasting, each subject was given one of the two test meals standardized, including 2 muffins and water (HSM, 42.1% of saturated fat , or HMM, 34.5% of monounsaturated fat ). Plasma fatty profile and concentrations of apolipoproteins A1 and B100, complement C3, and triacylglycerols were analyzed during fasting and at 2, 3, and 5 postprandial hours.Among the markers studied, the triacylglycerol (TAG) and complement C3 were significantly higher in the OW group, compared to NW. The increment in the C3 concentration was higher after HSM intake, compared with HMM (iAUC = 4365.5 ± 5477.4\xa0vs. 1215.2 ± 882.4; p = 0.006), with no differences between groups. After 5\xa0hours postprandial, plasma oleic values remained high compared with the fasting value in the NW group, but not in the OW group (26.0 ± 4.2\xa0vs 23.7 ± 3.9%; p < 0.001). Women with high percentage of total plasma saturated fatty acids (SFA) at the beginning of the intervention had higher incremental area under the curve (iAUC) for the , stearic, and total fatty acids (p < 0.005). Those women with a high percentage of monounsaturated fatty acids (MUFA) showed lower iAUC values for the same fatty profile (p < 0.005).This study demonstrated the effect of the HSM on postprandial increment of C3 concentration, suggesting another mechanism for saturated fat metabolism. The postprandial response to HSM appears to be the mediated by baseline lipid profile of the individuals, while the response to HMM was correlated to the weight status.

Keyword: obesity

Influence of Resveratrol on Sphingolipid Metabolism in Hepatocellular Carcinoma Cells in Lipid Overload State.

is characterized by increased long chain fatty acids (LCFA) uptake and impaired lipid metabolism in hepatocytes. Consequently, an enhanced intracellular lipid content, including sphingolipids, may lead to lipotoxicity. It is believed that resveratrol (RSV), one of the most extensively studied plant-derived polyphenols, and its interaction with sphingolipid metabolism may constitute one of the major therapeutic targets for cancer and metabolic diseases treatment.The aim of this study was to ascertain, whether resveratrol may affect sphingolipid metabolic pathways, enzymes and transporters in a lipid overload state.The experiments were conducted on hepatocellular carcinoma cells (HepG2) incubated with RSV and/or (PA) at the concentration of 0.5 mM and 50 µM, respectively for 16h. Intra- and extracellular sphingolipid concentrations were assessed by high-performance liquid chromatography and gas liquid chromatography. Moreover, the expression of caspase 3, selected fatty transporters and sphingolipid metabolism pathway proteins were estimated by Western Blot.RSV alone and together with PA significantly increased the intracellular concentration of ceramide, sphinganine and sphingosine as well as the expression of enzymes related to de novo ceramide synthesis pathway. Moreover, in our study, we observed augmented ceramide and sphingomyelin efflux into the incubation media in these groups. In addition, RSV substantially reduced intracellular triacylglycerols accumulation in lipid overload conditions.The above-mentioned findings suggest that RSV, at least partially, demonstrates a potential protective effect on HepG2 cells in a lipid overload state.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: obesity

-induced activation of JunD promotes myocardial lipid accumulation and metabolic cardiomyopathy.

Metabolic cardiomyopathy (MC)-characterized by intra-myocardial triglyceride (TG) accumulation and lipotoxic damage-is an emerging cause of heart failure in obese patients. Yet, its mechanisms remain poorly understood. The Activator Protein 1 (AP-1) member JunD was recently identified as a key modulator of hepatic lipid metabolism in obese mice. The present study investigates the role of JunD in -induced MC.JunD transcriptional activity was increased in hearts from diet-induced obese (DIO) mice and was associated with myocardial TG accumulation and left ventricular (LV) dysfunction. Obese mice lacking JunD were protected against MC. In DIO hearts, JunD directly binds PPARγ promoter thus enabling transcription of genes involved in TG synthesis, uptake, hydrolysis, and storage (i.e. Fas, Cd36, Lpl, Plin5). Cardiac-specific overexpression of JunD in lean mice led to PPARγ activation, cardiac steatosis, and dysfunction, thereby mimicking the MC phenotype. In DIO hearts as well as in neonatal rat ventricular myocytes exposed to , Ago2 immunoprecipitation, and luciferase assays revealed JunD as a direct target of miR-494-3p. Indeed, miR-494-3p was down-regulated in hearts from obese mice, while its overexpression prevented lipotoxic damage by suppressing JunD/PPARγ signalling. JunD and miR-494-3p were also dysregulated in myocardial specimens from obese patients as compared with non-obese controls, and correlated with myocardial TG content, expression of PPARγ-dependent genes, and echocardiographic indices of LV dysfunction.miR-494-3p/JunD is a novel molecular axis involved in -related MC. These results pave the way for approaches to prevent or treat LV dysfunction in obese patients.Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.

Keyword: obesity

Inhibition of central de novo ceramide synthesis restores insulin signaling in hypothalamus and enhances β-cell function of obese Zucker rats.

Hypothalamic lipotoxicity has been shown to induce central insulin resistance and dysregulation of glucose homeostasis; nevertheless, elucidation of the regulatory mechanisms remains incomplete. Here, we aimed to determine the role of de novo ceramide synthesis in hypothalamus on the onset of central insulin resistance and the dysregulation of glucose homeostasis induced by .Hypothalamic GT1-7 neuronal cells were treated with palmitate. De novo ceramide synthesis was inhibited either by pharmacological (myriocin) or molecular (si-Serine Palmitoyl Transferase 2, siSPT2) approaches. Obese Zucker rats (OZR) were intracerebroventricularly infused with myriocin to inhibit de novo ceramide synthesis. Insulin resistance was determined by quantification of Akt phosphorylation. Ceramide levels were quantified either by a radioactive kinase assay or by mass spectrometry analysis. Glucose homeostasis were evaluated in myriocin-treated OZR. Basal and glucose-stimulated parasympathetic tonus was recorded in OZR. Insulin secretion from islets and β-cell mass was also determined.We show that palmitate impaired insulin signaling and increased ceramide levels in hypothalamic neuronal GT1-7 cells. In addition, the use of deuterated demonstrated that palmitate activated several enzymes of the de novo ceramide synthesis pathway in hypothalamic cells. Importantly, myriocin and siSPT2 treatment restored insulin signaling in palmitate-treated GT1-7 cells. Protein kinase C (PKC) inhibitor or a dominant-negative PKCζ also counteracted palmitate-induced insulin resistance. Interestingly, attenuating the increase in levels of hypothalamic ceramides with intracerebroventricular infusion of myriocin in OZR improved their hypothalamic insulin-sensitivity. Importantly, central myriocin treatment partially restored glucose tolerance in OZR. This latter effect is related to the restoration of glucose-stimulated insulin secretion and an increase in β-cell mass of OZR. Electrophysiological recordings also showed an improvement of glucose-stimulated parasympathetic nerve activity in OZR centrally treated with myriocin.Our results highlight a key role of hypothalamic de novo ceramide synthesis in central insulin resistance installation and glucose homeostasis dysregulation associated with .Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: obesity

[The person in philogenesis is not (omnivores), but the herbivores with the carnivores past and the fuzzy future. Biological function of trophology (nutrition) in ontogenesis.]

According to the phylogenetic theory of general pathology, seven biological functions have been formed over billions of years. 1. biological function of trophology, nutrition; 2. homeostasis function; 3. biological function of endoecology; 4. function of adaptation; 5. function of the continuation of the species; 6. function of locomotion and 7. cognitive biological function, including intelligence. Millions of years in life consistently in the waters of several oceans, all the ancestors of man were carnivorous (Carnivores), fish-eating mammals. When the ocean retreated and the carnivorous (fish-eating) were on land, each individual privatized a "piece" of the ocean. Animals transformed it ito a pool of intercellular medium in vivo. The biological role of the late in the phylogeny of insulin is the formation of new biological functions in vivo. The action of insulin has transformed the carnivorous (fish-eating) ocean into herbivorous (Herbivores) species on land. There was it by synthesis in vivo from exogenous glucose of fatty acids (FA). Regulatory action of insulin was the directed conversion of exogenous glucose into ω-6 C18: 1 cis-oleic FA. Insulin late in phylogeny expressed the synthesis of new, conjugated enzymes: it is palmitoyl-CoA-elongase and stearyl-CoAdesaturase. Two enzymes synthesized FAs along the way: synthesized in situ de novo, from exogenous glucose, C16: 0 → C18: 0 stearic → ω-6 C18: 1 cis-oleic without accumulation of stearic FA. Insulin is not converted into an oleic FA exogenous from carnivorous food. On land, the action of insulin transformed the species Homo sapiens, into a herbivore, but with carnivorous, fish-eating, past. The idea of a person as omnivorous (Omnivor) - nonsense; such forms of nature did not form. Violation of the function of nutrition, the biological reaction of exotrophy (external nutrition), is the etiological and pathogenetic basis of the seven metabolic pandemics, the diseases of civilization. 1. Atherosclerosis and atheromatosis; 2. metabolic arterial hypertension; 3. metabolic syndrome; 4. ; 5. syndrome of insulin resistance; 6. non-alcoholic fatty liver disease and 7. endogenous hyperuricemia. The primary prevention of metabolic pandemics in the biological function of nutrition, in the biological reactions of exo-and endotrophy, will allow us to understand the theoretical bases and implementation of preventive actions that will determine the characteristics of nutrition in the future.

Keyword: obesity

Chitosan Oligosaccharides Improve Glucolipid Metabolism Disorder in Liver by Suppression of -Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid metabolism disorder using (PA)-induced HepG2 cells and high-fat diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated lipid accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid metabolism-related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid metabolism disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid metabolism-related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid metabolism disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid metabolism-related diseases.

Keyword: obesity

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: obesity

Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 Diabetes and in the Rat, as Suggested by the Effects on Stem Cell Culture.

Previous work showed that muscle-derived stem cells (MDSCs) exposed long-term to the milieu of uncontrolled type 2 diabetes (UC-T2D) in male obese Zucker (OZ) rats, were unable to correct the associated erectile dysfunction and the underlying histopathology when implanted into the corpora cavernosa, and were also imprinted with a noxious gene global transcriptional signature (gene-GTS), suggesting that this may interfere with their use as autografts in stem cell therapy.To ascertain the respective contributions of dyslipidemia and hyperglycemia to this MDSC damage, clarify its mechanism, and design a bioassay to identify the damaged stem cells.Early diabetes MDSCs and late diabetes MDSCs were respectively isolated from nearly normal young OZ rats and moderately hyperglycemic and severely dyslipidemic/obese aged rats with erectile dysfunction. Monolayer cultures of early diabetic MDSCs were incubated 4 days in DMEM/10% fetal calf serum\xa0+ or\xa0- aged OZ or lean Zucker serum from non-diabetic lean Zucker rats (0.5-5%) or with soluble (PA) (0.5-2 mM), cholesterol (CHOL) (50-400 mg/dL), or glucose (10-25 mM).Fat infiltration was estimated by Oil red O, apoptosis by TUNEL, protein expression by Western blots, and gene-GTS and microRNA (miR)-GTS were determined in these stem cells\' RNA.Aged OZ serum caused fat infiltration, apoptosis, myostatin overexpression, and impaired differentiation. Some of these changes, and also a proliferation decrease occurred with PA and CHOL. The gene-GTS changes by OZ serum did not resemble the in\xa0vivo changes, but some occurred with PA and CHOL. The miR-GTS changes by OZ serum, PA, and CHOL resembled most of the in\xa0vivo changes. Hyperglycemia did not replicate most alterations.MDSCs may be damaged in long-term UC-T2D/obese patients and be ineffective in autologous human stem cell therapy, which may be prevented by excluding the damaged MDSCs.The in\xa0vitro test of MDSCs is innovative and fast to define dyslipidemic factors inducing stem cell damage, its mechanism, prevention, and counteraction. Confirmation is required in other T2D/ rat models and stem cells (including human), as well as miR-GTS biomarker validation as a stem cell damage biomarker.Serum from long-term UC-T2D/obese rats or dyslipidemic factors induces a noxious phenotype and miR-GTS on normal MDSCs, which may lead in\xa0vivo to the repair inefficacy of late diabetic MDSCs. This suggests that autograft therapy with MDSCs in long-term UT-T2D obese patients may be ineffective, albeit this may be predictable by prior stem cell miR-GTS tests. Masouminia M, Gelfand R, Kovanecz I, et\xa0al. Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 Diabetes and in the Rat, as Suggested by the Effects on Stem Cell Culture. J Sex Med 2018;15:1678-1697.Copyright © 2018 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

Keyword: obesity

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing , hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced , hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: obesity

Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with metabolic syndrome.

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an energy-restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with metabolic syndrome.The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of metabolic syndrome.After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk energy-restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: obesity

Oleuropein improves insulin resistance in skeletal muscle by promoting the translocation of GLUT4.

As the beneficial effects of the Mediterranean diet on human health are well established, the phenolic compounds in olive oil have been gaining interest. Oleuropein, a major phenolic compound in olives, is known to reduce the blood glucose levels in alloxan-induced diabetic rats and rabbits, however, its effect on type 2 diabetes caused by is not clear. The purpose of this study is clarifying the effect of oleuropein on the glucose tolerance in skeletal muscle under the condition of lipotoxicity caused by type 2 diabetes. Oleuropein enhanced glucose uptake in C2C12 cells without insulin. Translocation of glucose transporter 4 (GLUT4) into the cell membrane was promoted by activation of adenosine monophosphate-activated protein kinase (AMPK) but not protein kinase B (Akt). Physiological concentration of oleuropein (10\xa0µM) was sufficient to express beneficial effects on C2C12 cells. Oleuropein prevented -induced myocellular insulin resistance. Furthermore, in gastrocnemius muscles of mice fed a high fat diet, oleuropein also induced the GLUT4 localization into cell membrane. These results suggest the possibility of oleuropein to be effective for type 2 diabetes by reducing insulin resistance in skeletal muscles.

Keyword: obesity

Rational design of dimeric lipidated Xenopus glucagon-like peptide 1 analogues as long-acting antihyperglycaemic agents.

Dimerization is viewed as an effective means to enhance the binding affinity and therapeutic potency of peptides. Both dimerization and lipidation effectively prolong the half-life of peptides in\xa0vivo by increasing hydrodynamic size and facilitating physical interactions with serum albumin. Here, we report a novel method to discover long-acting glucagon-like peptide 1 (GLP-1) analogues by rational design based on Xenopus GLP-1 through a combined dimerization and lipidization strategy. On the basis of our previous structure analysis of Xenopus GLP-1, and a C-terminal Cys were firstly introduced into two Xenopus GLP-1 analogues (1 and 2), and the afforded 3 and 4 were further reacted with bis-maleimide amine to afford two dimeric lipidated Xenopus GLP-1 analogues (5 and 6). The in\xa0vitro and in\xa0vivo biological activities of 5 and 6 were significantly improved as compared with their monomers. Moreover, the selected compound 6 showed greater hypoglycemic and insulinotropic activities than liraglutide even when the dose of 6 was reduced to half in db/db mice. Pharmacokinetic test revealed that 6 had a ∼ 3-fold longer half-life than liraglutide in Kunming mice and SD rats, and the longer half-life of 6 led to excellent long-acting hypoglycemic effects as confirmed by two different pharmacological methods conducted on db/db mice. Finally, a 7 weeks chronic study conducted on db/db mice demonstrated the better therapeutic efficacies of 6 on glucose tolerance normalization, HbA1c reduction and pancreas islets protection than liraglutide. The present research showed that combined dimerization and lipidization is effective when applied to Xenopus GLP-1 analogue to develop novel GLP-1 analogue for the treatment of type 2 diabetes. In addition, the promising preclinical data of 6 suggested the therapeutic potential of 6 as a novel anti-diabetic agent.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: obesity

Maternal overnutrition by hypercaloric diets programs hypothalamic mitochondrial fusion and metabolic dysfunction in rat male offspring.

Maternal overnutrition including pre-pregnancy, pregnancy and lactation promotes a lipotoxic insult leading to metabolic dysfunction in offspring. Diet-induced models (DIO) show that changes in hypothalamic mitochondria fusion and fission dynamics modulate metabolic dysfunction. Using three selective diet formula including a High fat diet (HFD), Cafeteria (CAF) and High Sugar Diet (HSD), we hypothesized that maternal diets exposure program leads to selective changes in hypothalamic mitochondria fusion and fission dynamics in male offspring leading to metabolic dysfunction which is exacerbated by a second exposure after weaning.We exposed female Wistar rats to nutritional programming including Chow, HFD, CAF, or HSD for 9\xa0weeks (pre-mating, mating, pregnancy and lactation) or to the same diets to offspring after weaning. We determined body weight, food intake and metabolic parameters in the offspring from 21 to 60\xa0days old. Hypothalamus was dissected at 60\xa0days old to determine mitochondria-ER interaction markers by mRNA expression and western blot and morphology by transmission electron microscopy (TEM). Mitochondrial-ER function was analyzed by confocal microscopy using hypothalamic cell line mHypoA-CLU192.Maternal programming by HFD and CAF leads to failure in glucose, leptin and insulin sensitivity and fat accumulation. Additionally, HFD and CAF programming promote mitochondrial fusion by increasing the expression of MFN2 and decreasing DRP1, respectively. Further, TEM analysis confirms that CAF exposure after programing leads to an increase in mitochondria fusion and enhanced mitochondrial-ER interaction, which partially correlates with metabolic dysfunction and fat accumulation in the HFD and CAF groups. Finally, we identified that lipotoxic stimulus in hypothalamic cells increases Ca overload into mitochondria matrix leading to mitochondrial dysfunction.We concluded that maternal programming by HFD induces hypothalamic mitochondria fusion, metabolic dysfunction and fat accumulation in male offspring, which is exacerbated by HFD or CAF exposure after weaning, potentially due to mitochondria calcium overflux.

Keyword: obesity

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism.A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit , hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: obesity

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: obesity

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of -related diseases.

Keyword: obesity

Cyclophilin D participates in the inhibitory effect of high-fat diet on the expression of steroidogenic acute regulatory protein.

The high-fat diet (HFD)-induced is responsible for the testosterone deficiency (TD). However, the mechanism remains unknown. Mitochondrial homeostasis is proved to be important for maintaining the function of steroidogenic acute regulatory protein (StAR), the first rate-limiting enzyme in testosterone synthesis. As the key regulator of mitochondrial membrane permeability, cyclophilin D (CypD) plays a crucial role in maintaining mitochondrial function. In this study, we sought to elucidate the role of CypD in the expression of StAR affected by HFD.To analyse the influence of CypD on StAR in vivo and in vitro, mouse models of HFD, CypD overexpression and CypD knockout (Ppif ) as well as Leydig cells treated with (PA) and CypD overexpression plasmids were examined with an array of metabolic, mitochondrial function and molecular assays.Compared with the normal diet mice, consistent with reduced testosterone in testes, the expressions of StAR in both mRNA and protein levels in HFD mice were down-regulated, while expressions of CypD were up-regulated. High-fat intake impaired mitochondrial function with the decrease in StAR in Leydig cells. Overexpression of CypD inhibited StAR expressions in vivo and in vitro. Compared with C57BL/6 mice with HFD, expressions of StAR were improved in Ppif mice with HFD.Mitochondrial CypD involved in the inhibitory effect of HFD on StAR expression in testes.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: obesity

Theaflavins Improve Insulin Sensitivity through Regulating Mitochondrial Biosynthesis in -Induced HepG2 Cells.

Theaflavins, the characteristic and bioactive polyphenols in black tea, possess the potential improving effects on insulin resistance-associated metabolic abnormalities, including and type 2 diabetes mellitus. However, the related molecular mechanisms are still unclear. In this research, we investigated the protective effects of theaflavins against insulin resistance in HepG2 cells induced by . Theaflavins significantly increased glucose uptake of insulin-resistant cells at noncytotoxic doses. This activity was mediated by upregulating the total and membrane bound glucose transporter 4 protein expressions, increasing the phosphor-Akt (Ser473) level, and decreasing the phosphorylation of IRS-1 at Ser307. Moreover, theaflavins were found to enhance the mitochondrial DNA copy number, down-regulate the PGC-1β mRNA level and increase the PRC mRNA expression. Mdivi-1, a selective mitochondrial division inhibitor, could attenuate TFs-induced promotion of glucose uptake in insulin-resistant HepG2 cells. Taken together, these results suggested that theaflavins could improve hepatocellular insulin resistance induced by free fatty acids, at least partly through promoting mitochondrial biogenesis. Theaflavins are promising functional food ingredients and medicines for improving insulin resistance-related disorders.

Keyword: obesity

Maternal results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether impacts placental lipid transport and metabolism remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and insulin-sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal . Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in lipid classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to insulin resistance and low-grade inflammation in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by .-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

Keyword: obesity

Nutritionally Derived Metabolic Cues Typical of the Obese Microenvironment Increase Cholesterol Efflux Capacity of Adipose Tissue Macrophages.

Cholesterol retention within plasma membranes of macrophages is associated with increased inflammatory signaling. Cholesterol efflux via the transporters ABCA1, ABCG1, and SR-BI to high-density lipoprotein (HDL) particles is a critical mechanism to maintain cellular cholesterol homeostasis. Little is known about the impact of the obese microenvironment on cholesterol efflux capacity (CEC) of macrophages. In this study, the CEC of obese-derived primary adipose-tissue macrophages (ATM) is evaluated and the in vivo microenvironment is modeled in vitro to determine mechanisms underlying modulated CEC.F4/80 ATM are labeled with H-cholesterol ex vivo, and CEC and ABCA1/ABCG1 protein levels are determined. Total, ABCA1-dependent, and ABCA1-independent CECs are determined in J774 macrophages polarized to M1 (LPS&IFNγ), M2 (IL-4&IL-13), or metabolic phenotypes (glucose, insulin, and ).Obese ATM exhibit enhanced CEC and ABCA1 and ABCG1 expression compared to lean ATM. In contrast, ABCA1-CEC is suppressed from M1 polarized macrophages compared to untreated in vitro, by activation of the JAK/STAT pathway. Incubation of macrophages in vitro in high glucose augments cAMP-induced ABCA1 protein expression and ABCA1-CEC.These novel findings demonstrate remarkable plasticity of macrophages to respond to their environment with specific modulation of ABCA1 depending on whether classical pro-inflammatory or metabolic cues predominate.© 2018 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: obesity

Effects of 1α,25 Dihydroxyvitamin D on Pro-inflammatory Cytokines of Treated Thp-1 Cells.

The level of saturated fatty acids, such as (PA), correlates with chronic inflammation in obese and metabolic syndrome patients. However, low level of vitamin D is observed in those conditions. The aim of this study is to investigate effects of 1α,25(OH) D on PA-treated THP-1 cells. Using quantitative real-time polymerase chain reaction, we measure mRNA expression of pro-inflammatory cytokines: TNF-α, Interleukin (IL)-1β, IL-6, and chemokine IL-8 under PA and 1α,25(OH) D influence. PA, at all concentrations (25-100 μM), enhanced LPS stimulatory effect on those mRNA expression compared to LPS-treated and -untreated cells. Combination with 1α,25(OH) D increased cytokine expression at high (10 M) and high-normal (10 M) concentrations compared to PA + LPS and LPS alone, both for 2 and 24 h. However, low-normal (10 M) and low (10 M) levels of 1α,25(OH) D could not enhance PA effect, but mRNA expression of pro-inflammatory cytokine was higher than LPS-treated cells. Upstream pathway of 1α,25(OH) D , which is cholecalciferol, also gave the similar result. Further, inhibition of calcium pathway does not play a role in this mechanism. Thus, these findings support pro-inflammatory effect of PA and vitamin D on innate immune response, especially on fat-induced inflammation.The effect of vitamin D on chronic inflammation in is uncertain. This study shows an in vitro possibility that vitamin D could exaggerate inflammation when combined with high SFAs. The idea of using vitamin D supplement to modulate inflammation in fat-related inflammation needs further refined experiments before its clinical application.© 2017 Institute of Food Technologists®.

Keyword: obesity

Exploring the interactions between serum free fatty acids and fecal microbiota in through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in . The gut microbiota is involved in the host energy metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting , showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: obesity

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid metabolism-related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced . Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of lipid metabolism- and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid metabolism-related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid metabolism, suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: obesity

induces neurotoxicity and gliatoxicity in SH-SY5Y human neuroblastoma and T98G human glioblastoma cells.

-related central nervous system (CNS) pathologies like neuroinflammation and reactive gliosis are associated with high-fat diet (HFD) related elevation of saturated fatty acids like (PA) in neurons and astrocytes of the brain.Human neuroblastoma cells SH-SY5Y (as a neuronal model) and human glioblastoma cells T98G (as an astrocytic model), were treated with 100-500 µM PA, oleic (OA) or lauric (LA) for 24 h or 48 h, and their cell viability was assessed by 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of stable overexpression of γ-synuclein (γ-syn), a neuronal protein recently recognized as a novel regulator of lipid handling in adipocytes, and transient overexpression of Parkinson\'s disease (PD) α-synuclein [α-syn; wild-type (wt) and its pathogenic mutants A53T, A30P and E46K] in SH-SY5Y and T98G cells, were also evaluated. The effects of co-treatment of PA with paraquat (PQ), a Parkinsonian pesticide, and leptin, a hormone involved in the brain-adipose axis, were also assessed. Cell death mode and cell cycle were analyzed by Annexin V/PI flow cytometry. Reactive oxygen species (ROS) level was determined using 2\',7\'-dichlorofluorescien diacetate (DCFH-DA) assay and lipid peroxidation level was determined using thiobarbituric reactive substances (TBARS) assay.MTT assay revealed dose- and time-dependent PA cytotoxicity on SH-SY5Y and T98G cells, but not OA and LA. The cytotoxicity was significantly lower in SH-SY5Y-γ-syn cells, while transient overexpression of wt α-syn or its PD mutants (A30P and E46K, but not A53T) modestly (but still significantly) rescued the cytotoxicity of PA in SH-SY5Y and T98G cells. Co-treatment of increasing concentrations of PQ exacerbated PA\'s neurotoxicity. Pre-treatment of leptin, an anti-apoptotic adipokine, did not successfully rescue SH-SY5Y cells from PA-induced cytotoxicity-suggesting a mechanism of PA-induced leptin resistance. Annexin V/PI flow cytometry analysis revealed PA-induced increase in percentages of cells in annexin V-positive/PI-negative quadrant (early apoptosis) and subG-G fraction, accompanied by a decrease in G-M phase cells. The PA-induced ROS production and lipid peroxidation was at greater extent in T98G as compared to that in SH-SY5Y.In conclusion, PA induces apoptosis by increasing oxidative stress in neurons and astrocytes. Taken together, the results suggest that HFD may cause neuronal and astrocytic damage, which indirectly proposes that CNS pathologies involving neuroinflammation and reactive gliosis could be prevented via the diet regimen.

Keyword: obesity

A high-fat diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high-fat diet (HFD) commonly leads to and insulin resistance. However, even 3\u2009days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus.Mice were fed either a low-fat diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from fat for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose metabolism and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: obesity

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and . Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: obesity

Saturated fatty acids promote chondrocyte matrix remodeling through reprogramming of autophagy pathways.

is a known strong risk factor for the onset of knee osteoarthritis and is often accompanied by dysregulated lipid metabolism with elevated levels of free fatty acids such as saturated fatty acids (SFAs). The purpose of this study was to determine how autophagy varies in chondrocytes in response to predominant SFAs such as lauric, myristic, , and stearic acids.Normal human articular cartilage chondrocytes and C28/I2 chondrocyte cell lines were stimulated with different SFAs in both the absence and presence of interleukin-1β to study the effects of SFA and inflammatory cytokines in mediating the activation of autophagy. The effects of rapamycin and LY290042 on autophagy in response to different SFAs were also assessed. and stearic stimulation of chondrocytes resulted in increased activation of both autophagy and the canonical NFκB pathway as evidenced by increased expressions of the key autophagy markers microtubule-associated protein-1 light chain 3, autophagy-related 5, beclin-1, and NFκB p65. In contrast, lauric stimulation resulted in decreased autophagy activation as shown by decreased expressions of microtubule-associated protein-1 light chain 3, autophagy-related 5, and beclin-1, which suggests decreased cellular stress.These results represent a novel mechanism by which various SFAs activate autophagy and simultaneously modulate NFκB signaling pathways and the expression of chondrocyte regulatory genes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

NDP-MSH reduces oxidative damage induced by in primary astrocytes.

Recent findings relate to inflammation in key hypothalamic areas for body weight control. Hypothalamic inflammation has also been related to oxidative stress. (PA) is the most abundant free fatty found in food, and in vitro studies indicate that it triggers a pro-inflammatory response in the brain. Melanocortins are neuropeptides with proven anti-inflammatory and neuroprotective action mediated by melanocortin receptor 4 (MC4R), but little is known about the effect of melanocortins on oxidative stress. The aim of this study was to investigate whether melanocortins could alleviate oxidative stress induced by a high fat diet (HFD) model. We found that NDP-MSH treatment decreased PA-induced reactive oxygen species production in astrocytes, an effect blocked by the MC4R inhibitor JKC363. NDP-MSH abolished nuclear translocation of Nrf2 induced by PA and blocked the inhibitory effect of PA on superoxide dismutase (SOD) activity and glutathione levels while it also per se increased activity of SOD and γ-glutamate cysteine ligase (γ-GCL) antioxidant enzymes. However, HFD reduced hypothalamic MC4R and brain derived neurotrophic factor mRNA levels, thereby preventing the neuroprotective mechanism induced by melanocortins.© 2019 British Society for Neuroendocrinology.

Keyword: obesity

The inhibition of Nrf2 accelerates renal lipid deposition through suppressing the ACSL1 expression in -related nephropathy.

has become a worldwide epidemic, and the incidence of is increasing year by year. -related nephropathy (ORN) is a common kidney complication of . Long-chain acyl-CoA synthetases-1, (ACSL1), is a key enzyme in the oxidative metabolism of fatty acids in mitochondria and ACSL1 may play a direct role in renal lipid deposition and promote the progress of ORN. In this study, we focus on the renoprotective role of ACSL1 in ORN. Electron microscopy, immunohistochemical (IHC) staining, Western blot, and real-time PCR were used to detect the expression of ACSL1and Nrf2 in ORN patients, ob/ob mice and (PA)-treated HK-2 cells. Oil red staining and Elisa Kit were used to detect the intracellular FFA and TG contents in ob/ob mice and PA-treated HK-2 cells. Dihydroethidium (DHE) staining and the MDA/SOD measurement were used to detect the ROS production. In order to demonstrate the role of ACSL1 and the interaction between ACSL1 and Nrf2 in ORN, related siRNA and plasmid were transfected into HK-2 cells. More ROS production and renal lipid deposition have been found in ORN patients, ob/ob mice and PA-treated HK-2 cells. Compared with control, all the expression of ACSL1and Nrf2 were down-regulated in ORN patients, ob/ob mice and PA-treated HK-2 cells. The Nrf2 could regulate the expression of ACSL1 and the ACSL1 played the direct role in renal lipid deposition. The Nrf2 is inhibited in ORN, resulting more ROS production and oxidative stress. Increased oxidative stress will suppress the expression of ACSL1, which could increase the intracellular FFA and TG contents, ultimately leading to renal lipid deposition in renal tubulars and accelerating the development of ORN.

Keyword: obesity

Inhibition of soluble epoxide hydrolase attenuates a high-fat diet-mediated renal injury by activating PAX2 and AMPK.

A high-fat diet (HFD) causes -associated morbidities involved in macroautophagy and chaperone-mediated autophagy (CMA). AMPK, the mediator of macroautophage, has been reported to be inactivated in HFD-caused renal injury. However, PAX2, the mediator for CMA, has not been reported in HFD-caused renal injury. Here we report that HFD-caused renal injury involved the inactivation of Pax2 and Ampk, and the activation of soluble epoxide hydrolase (sEH), in a murine model. Specifically, mice fed on an HFD for 2, 4, and 8 wk showed time-dependent renal injury, the significant decrease in renal Pax2 and Ampk at both mRNA and protein levels, and a significant increase in renal sEH at mRNA, protein, and molecular levels. Also, administration of an sEH inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea, significantly attenuated the HFD-caused renal injury, decreased renal sEH consistently at mRNA and protein levels, modified the renal levels of sEH-mediated epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) as expected, and increased renal Pax2 and Ampk at mRNA and/or protein levels. Furthermore, (PA) treatment caused significant increase in , and decrease in both Pax2 and Ampk in murine renal mesangial cells (mRMCs) time- and dose-dependently. Also, 14(15)-EET (a major substrate of sEH), but not its sEH-mediated metabolite 14,15-DHET, significantly reversed PA-induced increase in , and PA-induced decrease in Pax2 and Ampk. In addition, plasmid construction revealed that Pax2 may positively regulate Ampk transcriptionally in mRMCs. This study provides insights into and therapeutic target for the HFD-mediated renal injury.

Keyword: obesity

Tibolone Reduces Oxidative Damage and Inflammation in Microglia Stimulated with through Mechanisms Involving Estrogen Receptor Beta.

High concentrations of in plasma increase both the inflammation associated with and the susceptibility to develop a neurodegenerative event. In the brain, the inflammatory response is mediated by activated microglial cells, which undergo morphological and biochemical changes and can directly affect cell viability. Recent evidence shows that the use of estrogenic compounds can control microglia-induced inflammation with promising results. In this study, we explored the actions of the synthetic steroid tibolone on BV-2 microglia cells stimulated with . Our results demonstrated that tibolone increased cell viability and reduced nuclear fragmentation and the production of reactive oxygen species, as well as preserved mitochondrial membrane potential. These effects were accompanied by reduced nuclear translocation of NF-κB p65, upregulation of neuroglobin, and improved antioxidant defense. Furthermore, estrogen receptor beta (ERβ) inhibition partially dampened tibolone\'s protective actions in BV-2 cells stimulated with . In conclusion, tibolone protects BV-2 cells by a mechanism involving ERβ and upregulation of neuroglobin.

Keyword: obesity

Hoxa5 increases mitochondrial apoptosis by inhibiting Akt/mTORC1/S6K1 pathway in mice white adipocytes.

Homeobox A5(), a member of the family, plays a important role in the regulation of proliferation and apoptosis in cancer cells. The dysregulation of the adipocyte apoptosis leads to and metabolic disorders. However, the effects of Hoxa5 on adipocyte apoptosis are still unknown. In this study, (PA) significantly increased the mRNA level of and triggered white adipocyte apoptosis and . Further analysis revealed that Hoxa5 enhanced the early and late apoptotic cells and fragmentation of genomic DNA in adipocytes from inguinal white adipose tissue (iWAT) of mice. Moreover, Hoxa5 aggravated white adipocyte apoptosis through mitochondrial pathway rather than endoplasmic reticulum stress (ERS)-induced or death receptor (DR)-mediated pathway. Our data also confirmed that Hoxa5 promoted mitochondrial apoptosis pathway by elevating the transcription activity of and inhibiting the protein kinase B (Akt)/mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. In summary, these findings revealed a novel mechanism that linked Hoxa5 to white adipocyte apoptosis, which provided some potential possibilities to prevent and treat and some metabolic diseases.

Keyword: obesity

Resveratrol prevents -induced cardiomyocyte contractile impairment.

Long-chain saturated fatty acids, especially (PA), contribute to cardiomyocyte lipotoxicity. This study tests the effects of PA on adult rat cardiomyocyte contractile function and proteins associated with calcium regulating cardiomyocyte contraction and relaxation. Adult rat cardiomyocytes were pretreated with resveratrol (Resv) and then treated with PA. For the reversal study, cardiomyocytes were incubated with PA prior to treatment with Resv. Cardiomyocyte contractility, ratio of rod- to round-shaped cardiomyocytes, and Hoechst staining were used to measure functional and morphological changes in cardiomyocytes. Protein expression of sarco-endoplasmic reticulum ATPase 2a (SERCA2a), native phospholamban (PLB) and phosphorylated PLB (pPLB ser16 and pPLB thr17), and troponin I (TnI) and phosphorylated TnI (pTnI) were measured. SERCA2a activity was also measured. Our results show that PA (200 μM) decreased the rate of cardiomyocyte relaxation, reduced the number of rod-shaped cardiomyocytes, and increased the number of cells with condensed nuclei; pre-treating cardiomyocytes with Resv significantly prevented these changes. Post-treatment with Resv did not reverse morphological changes induced by PA. Protein expression levels of SERCA2a, PLB, pPLBs, TnI, and pTnI were unchanged by PA or Resv. SERCA2a activity assay showed that and Iono ratio were increased with PA and pre-treatment with Resv prevented this increase. In conclusion, our results show that Resv protect cardiomyocytes from contractile dysfunction induced by PA.

Keyword: obesity

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high fat diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of and -related metabolic diseases such as type II diabetes mellitus (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: obesity

Activator protein-1 and caspase 8 mediate p38α MAPK-dependent cardiomyocyte apoptosis induced by .

Lipoapoptosis of cardiomyocytes may underlie diabetic cardiomyopathy. Numerous forms of cardiomyopathies share a common end-pathway in which apoptotic loss of cardiomyocytes is mediated by p38α mitogen activated protein kinase (MAPK). Although we have previously shown that (PA), a saturated fatty (SFA) elevated in plasma of type 2 diabetes mellitus and morbid , induces apoptosis in cardiomyocytes via p38α MAPK-dependent signaling, the downstream cascade events that cause cell death remain unknown. The objective of this study\xa0was to investigate mechanisms involved in -induced cardiomyocyte apoptosis. Human adult ventricular cardiomyocyte line (AC16 cells) exposed to high physiological levels of PA for 16\xa0h showed enhanced transcription and phosphorylation of c-fos and c-jun subunits of AP-1 and transcription of caspase 8. When AC16 cells were transfected with small interfering RNA specific against p38α MAPK (si-p38α) for 24 or 48\xa0h, the amplified phosphorylation of c-fos was dose-dependently attenuated, and procaspase 8 was dose-dependently reduced. With translational knockdown of c-fos, PA-induced apoptosis was diminished. Inhibition of caspase 8 for 24\xa0h reduced apoptosis in PA-treated cardiomyocytes. These findings provide evidence for induction of apoptosis in cardiomyocytes exposed to high SFA by a novel pathway requiring activation of c-fos/AP-1 and caspase 8. These results demonstrate how elevated plasma SFA may lead to continual and cumulative loss of cardiomyocytes and potentially contribute to the\xa0development of diabetic cardiomyopathy.

Keyword: obesity

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/lipid metabolism in type 2 diabetes mellitus. Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high-fat diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for and its related T2DM.

Keyword: obesity

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced .

is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of ; however, its role in the metabolic consequences of is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with . In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent -associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: obesity

METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as , insulin resistance, and diabetes. Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: obesity

Fatty acids, epigenetic mechanisms and chronic diseases: a systematic review.

Chronic illnesses like , type 2 diabetes (T2D) and cardiovascular diseases, are worldwide major causes of morbidity and mortality. These pathological conditions involve interactions between environmental, genetic, and epigenetic factors. Recent advances in nutriepigenomics are contributing to clarify the role of some nutritional factors, including dietary fatty acids in gene expression regulation. This systematic review assesses currently available information concerning the role of the different fatty acids on epigenetic mechanisms that affect the development of chronic diseases or induce protective effects on metabolic alterations.A targeted search was conducted in the PubMed/Medline databases using the keywords "fatty acids and epigenetic". The data were analyzed according to the PRISMA-P guidelines.Consumption fatty acids like n-3 PUFA: EPA and DHA, and MUFA: oleic and palmitoleic was associated with an improvement of metabolic alterations. On the other hand, fatty acids that have been associated with the presence or development of , T2D, pro-inflammatory profile, atherosclerosis and IR were n-6 PUFA, saturated fatty acids (stearic and ), and trans fatty acids (elaidic), have been also linked with epigenetic changes.Fatty acids can regulate gene expression by modifying epigenetic mechanisms and consequently result in positive or negative impacts on metabolic outcomes.

Keyword: obesity

Anxa2 gene silencing attenuates -induced insulin resistance by suppressing the NF-κB signaling pathway.

Insulin resistance (IR) continues to pose a major threat to public health due to its role in the pathogenesis of metabolic syndrome and its ever-increasing prevalence on a global scale. The aim of the current study was to investigate the efficacy of Anxa2 in -induced IR through the mediation of the NF-κB signaling pathway. Microarray analysis was performed to screen differentially expressed genes associated with . To verify whether Anxa2 was differentially expressed in IR triggered by , IR mouse models were established in connection with a high-fat diet (HFD). In the mouse IR model, the role of differentially expressed Anxa2 in glycometabolism and IR was subsequently detected. To investigate the effect of Anxa2 on IR and its correlation with inflammation, a (PA)-induced IR cell model was established, with the relationship between Anxa2 and the NF-κB signaling pathway investigated accordingly. Anxa2 was determined to be highly expressed in IR. Silencing Anxa2 was shown to inhibit IR triggered by . When Anxa2 was knocked down, elevated expression of phosphorylated insulin receptor substrate 1 (IRS1), IRS1 and peroxisome proliferator-activated receptor coactivator-1a, and glucose tolerance and insulin sensitivity along with 2-deoxy-d-glucose uptake was detected, whereas decreased expression of suppressor of cytokine signaling 3, IL-6, IL-1β, TNF-α, and p50 was observed. Taken together, the current study ultimately demonstrated that Anxa2 may be a novel drug strategy for IR disruption, indicating that Anxa2 gene silencing is capable of alleviating PA or HFD-induced IR and inflammation through its negative regulatory role in the process of p50 nuclear translocation of the NF-κB signaling pathway.

Keyword: obesity

Adipocyte-Derived Exosomal MiR-27a Induces Insulin Resistance in Skeletal Muscle Through Repression of PPARγ.

The mechanism by which adipocyte-derived endocrine factors promote insulin resistance in skeletal muscle are not fully understood. MiR-27a is highly expressed in sera of obese individuals with prediabetes and T2DM, and mainly derived by adipose tissues. Thus, miR-27a secreted into circulation by adipose tissue may regulate insulin resistance in skeletal muscle. The association between miR-27a and insulin resistance in skeletal muscle was determined in obese children, high-fat diet-induced miR-27a knockdown obese mice, db/db mice and C2C12 cells overexpressing miR-27a. The crosstalk mediated by exosomal miR-27a between adipose tissue and skeletal muscle was determined in C2C12 cells incubated with conditioned medium prepared from palmitate-treated 3T3-L1 adipocytes. We showed that serum miR-27a level correlated positively with and insulin resistance in obese children, and that elevated serum miR-27a levels correlated with insulin resistance in leptin receptor-deficient db/db mice, and with and insulin resistance in high-fat diet-fed C57BL/6J mice. MiR-27a released from adipocytes of high-fat diet-fed C57BL/6J mice was associated with triglyceride accumulation. MiR-27a derived from these adipocytes induced insulin resistance in C2C12 skeletal muscle cells through miR-27a-mediated repression of PPARγ and its downstream genes involved in the development of . These results identify a novel crosstalk signaling pathway between adipose tissue and skeletal muscle in the development of insulin resistance, and indicate that adipose tissue-derived miR-27a may play a key role in the development of -triggered insulin resistance in skeletal muscle.

Keyword: obesity

A microplate assay for measuring cell death in C2C12 cells.

The main goal of this study was to develop a straightforward and rapid microplate assay for measuring propidium iodide (PI) in C2C12 cells. The PI method has proven to be an efficient quantitative assay for analyzing cell viability through PI fluorescence analysis. Importantly, the protocol takes less than 30 min and the results are reproducible. C2C12 cells were exposed to an increasing concentration of palmitate for a period of 24 h to induce cell death, and the PI fluorescence increased in a concentration-dependent manner. Evaluation of mitochondrial function and the production of reactive oxygen species confirmed the deleterious effects of palmitate. Also, the microplate PI assay demonstrated high sensitivity, as indicated by the detection of modest fluctuations in cell viability in response to catalase overexpression in palmitate-treated cells. The microplate PI assay, therefore, offers an accurate method for use in in-vitro studies.

Keyword: obesity

Saturated fatty acids bound to albumin enhance osteopontin expression and cleavage in renal proximal tubular cells.

Osteopontin (OPN) is one of the proinflammatory cytokines upregulated in the kidneys of diabetic animals and patients with nephropathy. An increase in urinary albumin and albumin-bound fatty acids (FA) presents a proinflammatory environment to the proximal tubules in proteinuric kidney diseases including diabetic nephropathy. This study was designed to examine if FA overload could stimulate OPN expression and cleavage in renal tubule epithelial cells. OPN gene and protein expression was examined in the kidney of Zucker diabetic (ZD) rats and cultured proximal tubular cells exposed to either bovine serum albumin (BSA) or BSA conjugated with (PA), the most abundant saturated plasma FA. Real-time PCR analysis confirmed an upregulation of renal cortical OPN gene correlated with albuminuria and nephropathy progression in ZD rats at the age of 7-20 weeks. Immunofluorescence staining of kidney sections revealed a massive induction of OPN protein in albumin-overloaded proximal tubules of ZD rats. A significant increase in both intact and cleaved OPN proteins was further demonstrated in the diabetic kidney and urine samples, which was attenuated by antiproteinuric treatment with losartan, an angiotensin II receptor blocker. When exposed to fatty -free BSA, NRK-52E cells exhibited an increase in protein levels of full-length and cleaved OPN. Moreover, the increase in OPN fragments was greatly enhanced in the presence of PA (250-500 µM). Together, our results support a stimulatory effect of albumin and conjugated FA on OPN expression and cleavage in renal tubule epithelial cells. Thus, besides lowering albuminuria/proteinuria, mitigating circulating FAs may be an effective intervention for preventing and slowing down the progression of nephropathy associated with and type 2 diabetes.

Keyword: obesity

PINK1-Parkin alleviates metabolic stress induced by in adipose tissue and in 3T3-L1 preadipocytes.

Mitochondria play an important role in cellular metabolism and are closely related with metabolic stress. Recently, several studies have shown that mitophagy mediated by PTEN-induced putative kinase 1 (PINK1) and Parkin may play a critical role in clearing the damaged mitochondria and maintaining the overall balance of intracellular mitochondria in quality and quantity. A previous study showed that PINK1 and Parkin were overexpressed in adipose tissue in obese subjects. However, it is still unclear whether a direct relationship exists between and mitophagy. In this study, we created a high-fat-diet (HFD)-induced obese mouse model and examined the expression of PINK1 and Parkin in adipose tissue using western blot and real-time quantitative PCR. After we confirmed that there is an interesting difference between regular-chow-fed mice and HFD-induced obese mice in the expression of PINK1 and Parkin in\xa0vivo, we further tested the expression of PINK1 and Parkin in 3T3-L1 preadipocytes in\xa0vitro by treating cells with (PA) to induce metabolic stress. To better understand the role of PINK1 and Parkin in metabolic stress, 3T3-L1 preadipocytes were transfected with small interfering RNA (siRNA) of PINK1 and Parkin followed by PA treatment. Our results showed that under lower concentrations of PA, PINK1 and Parkin can be activated and play a protective role in resisting the harmful effects of PA, including protecting the mitochondrial function and resisting cellular death, while under higher concentrations of PA, the expression of PINK1 and Parkin can be inhibited. These results suggest that PINK1-Parkin can protect mitochondrial function against metabolic stress induced by or PA to a certain degree.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

ALDH2 protects against high fat diet-induced cardiomyopathy and defective autophagy: role of CaM kinase II, histone H3K9 methyltransferase SUV39H, Sirt1, and PGC-1α deacetylation.

Uncorrected contributes to cardiac remodeling and contractile dysfunction although the underlying mechanism remains poorly understood. Mitochondrial aldehyde dehydrogenase (ALDH2) is a mitochondrial enzyme with some promises in a number of cardiovascular diseases. This study was designed to evaluate the impact of ALDH2 on cardiac remodeling and contractile property in high fat diet-induced .Wild-type (WT) and ALDH2 transgenic mice were fed low (10% calorie from fat) or high (45% calorie from fat) fat diet for 5 months prior to the assessment of cardiac geometry and function using echocardiography, IonOptix system, Lectin, and Masson Trichrome staining. Western blot analysis was employed to evaluate autophagy, CaM kinase II, PGC-1α, histone H3K9 methyltransferase SUV39H, and Sirt-1.Our data revealed that high fat diet intake promoted weight gain, cardiac remodeling (hypertrophy and interstitial fibrosis, p\u2009<\u20090.0001) and contractile dysfunction (reduced fractional shortening (p\u2009<\u20090.0001), cardiomyocyte function (p\u2009<\u20090.0001), and intracellular Ca handling (p\u2009=\u20090.0346)), mitochondrial injury (elevated O levels, suppressed PGC-1α, and enhanced PGC-1α acetylation, p\u2009<\u20090.0001), elevated SUV39H, suppressed Sirt1, autophagy and phosphorylation of AMPK and CaM kinase II, the effects of which were negated by ALDH2 (p\u2009≤\u20090.0162). In vitro incubation of the ALDH2 activator Alda-1 rescued against -induced changes in cardiomyocyte function, the effect of which was nullified by the Sirt-1 inhibitor nicotinamide and the CaM kinase II inhibitor KN-93 (p\u2009<\u20090.0001). The SUV39H inhibitor chaetocin mimicked Alda-1-induced protection again (p\u2009<\u20090.0001). Examination in overweight human revealed an inverse correlation between diastolic cardiac function and ALDH2 gene mutation (p\u2009<\u20090.05).Taken together, these data suggest that ALDH2 serves as an indispensable factor against cardiac anomalies in diet-induced through a mechanism related to autophagy regulation and facilitation of the SUV39H-Sirt1-dependent PGC-1α deacetylation.

Keyword: obesity

Apigenin Ameliorates the -Induced Skeletal Muscle Atrophy by Attenuating Mitochondrial Dysfunction in the Muscle of Obese Mice.

It was investigated whether apigenin (AP) protected against skeletal muscle atrophy induced by .Mice were fed a high-fat diet (HFD) for 9 weeks to induce , and then were assigned to two groups; the HFD group received a high-fat diet, and the HFD+AP group received a 0.1% AP-containing HFD. After additional feeding of the experimental diet for 8 weeks, mice in the HFD group were highly obese compared with the mice in the standard diet fed mice group. The mice in the AP-treated group showed less fat pad accumulation and less inflammatory cytokines without body weight reduction. The weight of skeletal muscle in the AP group tended to increase as compared with that of the HFD group. Furthermore, AP reduced the expression of atrophic genes, including MuRF1 and Atrogin-1, but increased the exercise capacity. The mitochondrial function and mitochondrial biogenesis were enhanced by AP. In cultured C2C12 cells, AP also suppressed -induced muscle atrophy and mitochondrial dysfunction. In addition, AP activated AMP-activated protein kinase (AMPK) in the C2C12 and the muscle of HFD-induced obese mice.The results suggested that AP ameliorated the -induced skeletal muscle atrophy by attenuating mitochondrial dysfunction.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: obesity

Dietary modulation of energy homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of , type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy metabolism is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate , T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: obesity

Antihyperlipidemic Effect, Identification and Isolation of the Lipophilic Components from Artemisia integrifolia.

L (Compositae) is a medicinal and edible plant. To investigate its antihyperlipidemic effect, a crude lipophilic extract and the composing compounds were isolated and fractioned from the petroleum ether extract of aerial parts of using column chromatography on silica gel. The anti-hyperlipidemia effect was studied in a rat model of acute hyperlipidemia, which was induced by triton WR-1339. A new compound, integrinol (), together with nine known compounds, namely chamazulene (), acetylenes (E)-2 (), acetylenes (E)-3 (), eugenol (), (), oleic (), linoleic (), linolenic () and 12,13-epoxylinolenic were isolated from the crude lipophilic extract of A. integrifolia. The LD50 value of the crude extract was more than 4g/kg. In Triton WR-1339-induced acute hyperlipidemia model, the crude lipophilic extract (200 mg/kg) significantly reduced total cholesterol (TC) by 70% ( ≤ 0.01) and triglycerides (TGs) by 94% ( ≤ 0.001). The fractioned compounds, such as chamazulene (1), acetylene-2 (2), and linolenic (9), used at 4 mg/kg dose, also significantly decreased the concentrations of TC (32%, 33% and 64%, respectively) and TGs (48%, 33% and 93%, respectively). These compounds (i.e., chamazulene, acetylenes (E)-2, and linolenic ) were considered to be responsible for the bioactive antihyperlipidemic effect. In conclusion, the crude lipid extract of Artemisia integrifolia L could be used as a potential treatment to avert hyperlipidemia. Further studies to confirm these results in other models of hyperlipidemia (e.g., diet-induced ) are warranted.

Keyword: obesity

Pioglitazone Enhances Cytosolic Lipolysis, β-oxidation and Autophagy to Ameliorate Hepatic Steatosis.

Non-alcoholic fatty liver disease closely contributes to the development of and insulin resistance. Even though pioglitazone has been reported to effectively lessen hepatic steatosis in human studies, its molecular mechanism remains unclear. This study is designed to investigate the regulation of cytosolic lipolysis, β-oxidation and autophagy by pioglitazone in a mice model of high fat diet (HFD) and cell model incubated with . Our results revealed hepatic steatosis was apparently induced by HFD and it was significantly reversed by pioglitazone. The serum insulin and hepatic triglyceride content was significantly decreased by co-administered pioglitazone with HFD. Hepatic expression of cytosolic-lipolysis related proteins (ATGL, HSL), β-oxidation (CPT-1A) and autophagy-related proteins (ATG7, LC3, LAL) was significantly enhanced by pioglitazone. Knockdown PPARα/PPARγ in AML12 cells significantly and proportionally reduced the expressions of ATGL, CPT-1A and LC3II, which was induced by pioglitazone. Furthermore, facilitation of the autophagic flux by pioglitazone was obviously blocked by lysosomal inhibitor, leupeptin, to demonstrate accumulation of the LC3II and intracellular lipid in AML12 cells. Our results demonstrated that pioglitazone attenuating the hepatic steatosis may be mediated by enhancing cytosolic lipolysis, β-oxidation and autophagy in a PPARα and PPARγ dependent manner.

Keyword: obesity

Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration.

may promote intervertebral disc degeneration (IDD) by non-mechanical means, by influencing levels of free fatty acids which could impair cell metabolism. This study aims to establish metabolic factors in -related IDD independent of mechanical loading. In clinical study, we retrospectively reviewed 128 volunteers (73 males, 55 females, aged 29-88 years) and compared their grades of disk degeneration with -related factors such as body weight, BMI, and serum lipid levels. Clinically, the IDD group showed increased age, BMI and serum triglyceride. Triglyceride was a significant risk factor for IDD even after correction for BMI and age (P = 0.007). In animal model, rats were fed a high-fat diet (HFD) in order to study its effects on disk metabolism and apoptosis. HFD rats had significantly higher serum levels of lipids, including triglyceride and non-esterified fatty , and showed significantly decreased markers of anabolism, increased catabolism and apoptosis in disk. Finally, rat nucleus pulposus (NP) cells were stimulated with a fatty (, PA) to gauge its effects on cell metabolism and apoptosis. Cell culture studies showed that NP cells exposed to PA showed increased apoptosis for activation of caspase 3, 7, 9, and PARP, which was primarily via the MAPK signal pathway, especially ERK pathway. In conclusion, hypertriglyceridemia can lead to IDD, independently of age and BMI. Hypertriglyceridemia appears to mediate disk cell apoptosis and matrix catabolism primarily via the ERK pathway.Copyright © 2019 Zhang, Chen, Huang, Wang, Shan, Liu, Chen, Li, Fan and Zhao.

Keyword: obesity

Pex11a deficiency causes dyslipidaemia and in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and .© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: obesity

Amelioration of Endoplasmic Reticulum Stress by Mesenchymal Stem Cells via Hepatocyte Growth Factor/c-Met Signaling in -Associated Kidney Injury.

Recent advances in the understanding of lipid metabolism suggest a critical role of endoplasmic reticulum (ER) stress in -induced kidney injury. Hepatocyte growth factor (HGF) is a pleiotropic cytokine frequently featured in stem cell therapy with distinct renotropic benefits. This study aims to define the potential link between human induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs)/bone marrow-derived MSCs (BM-MSCs) and ER stress in lipotoxic kidney injury induced by (PA) in renal tubular cells and by high-fat diet (HFD) in mice. iPS-MSCs or BM-MSCs alleviated ER stress (by preventing induction of Bip, chop, and unfolded protein response), inflammation (Il6, Cxcl1, and Cxcl2), and apoptosis (Bax/Bcl2 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells) in renal cortex of animals exposed to HFD thus mitigating histologic damage and albuminuria, via activating HGF/c-Met paracrine signaling that resulted in enhanced HGF secretion in the glomerular compartment and c-Met expression in the tubules. Coculture experiments identified glomerular endothelial cells (GECs) to be the exclusive source of glomerular HGF when incubated with either iPS-MSCs or BM-MSCs in the presence of PA. Furthermore, both GEC-derived HGF and exogenous recombinant HGF attenuated PA-induced ER stress in cultured tubular cells, and this effect was abrogated by a neutralizing anti-HGF antibody. Taken together, this study is the first to demonstrate that MSCs ameliorate lipotoxic kidney injury via a novel microenvironment-dependent paracrine HGF/c-Met signaling mechanism to suppress ER stress and its downstream pro-inflammatory and pro-apoptotic consequences. Stem Cells Translational Medicine 2019;8:898&910.© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

Keyword: obesity

Low molecular weight fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic fatty-liver disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common liver disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in diabetes-induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the liver of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for /diabetes-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti- remedy. Several pharmacological studies have shown its beneficial effects in metabolism. Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate lipid and glucose metabolism and are therapeutic targets for type 2 diabetes. This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, lipid accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased lipid accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: obesity

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic insulin resistance development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Fatty Acids Consumption: The Role Metabolic Aspects Involved in and Its Associated Disorders.

and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut microbiota, and hepatic metabolism.

Keyword: obesity

Lipotoxic Effects of on Astrocytes Are Associated with Autophagy Impairment.

is associated with an increase in the brain levels of saturated free fatty acids, such as (PA). Previous studies have shown that PA exerts proinflammatory actions and reduces cell viability in astrocyte cultures. In this study, we have assessed whether an alteration in autophagy is involved in the effects of PA on astrocytes. Primary astrocytes were obtained from the cerebral cortex of male and female CD1 mouse pups and were incubated for 4.5 or 24\xa0h with 250-500\xa0μM PA. PA increased the levels of LC3-II, an autophagosome marker, and reduced LC3-II flux in astrocytes, suggesting a blockade of autophagy. This effect was observed both after 4.5 and 24\xa0h of treatment with PA. PA had additional effects after treatment for 24\xa0h, increasing the expression of proinflammatory cytokines, decreasing cell viability, and increasing the levels of an endoplasmic reticulum stress marker. In addition, PA decreased the expression of estrogen receptors, but only in female astrocytes. However, the treatment with estradiol, estrogen receptor agonists, or inhibitor of estradiol synthesis did not counteract the action of PA on cell viability. Rapamycin, an autophagy inducer, was unable to prevent the effect of PA on cell viability. In addition, hydroxychloroquine, an autophagy blocker, did not cause per se astrocyte death. These findings suggest that the effect of PA on autophagy is not sufficient to induce astrocyte loss, which is only observed when prolonged PA treatment causes other alterations in astrocytes, such as increased inflammation and endoplasmic reticulum stress.

Keyword: obesity

Palmitate-Induced Insulin Hypersecretion and Later Secretory Decline Associated with Changes in Protein Expression Patterns in Human Pancreatic Islets.

In obese children with high circulating concentrations of free fatty palmitate, we have observed that insulin levels at fasting and in response to a glucose challenge were several times higher than in obese children with low concentrations of the fatty as well as in lean controls. Declining and even insufficient insulin levels were observed in obese adolescents with high levels of the fatty . In isolated human islets exposed to palmitate we have observed insulin hypersecretion after 2 days exposure. In contrast, insulin secretion from the islets was reduced after 7 days culture in the presence of the fatty . This study aims\xa0at identifying islet-related biological events potentially linked with the observed insulin hypersecretion and later secretory decline in these obese children and adolescents using the islet model. We analyzed protein expression data obtained from human islets exposed to elevated palmitate levels for 2 and 7 days by an improved methodology for statistical analysis of differentially expressed proteins. Protein profiling of islet samples by liquid chromatography-tandem mass spectrometry identified 115 differentially expressed proteins (DEPs). Several DEPs including sorcin were associated with increased glucose-stimulated insulin secretion\xa0in islets after 2 days of exposure to palmitate. Similarly, several metabolic pathways including altered protein degradation, increased autophagy, altered redox condition, and hampered insulin processing were coupled to the functional impairment of islets after 7 days of culture in the presence of palmitate. Such biological events, once validated in the islets, may give rise to novel treatment strategies aiming at normalizing insulin levels in obese children with high palmitate levels, which may reduce or even prevent -related type 2 diabetes mellitus.

Keyword: obesity

Overexpression of heart-type fatty binding protein enhances fatty -induced podocyte injury.

Deregulated lipid metabolism is a characteristic of metabolic diseases including type 2 diabetes and , and likely contributes to podocyte injury and end-stage kidney disease. Heart-type fatty binding protein (H-FABP) was reported to be associated with lipid metabolism. The present study investigated whether H-FABP contributes to podocyte homeostasis. Podocytes were transfected by lentiviral vector to construct a cell line which stably overexpressed H-FABP. Small interfering RNA capable of effectively silencing H-FABP was introduced into podocytes to construct a cell line with H-FABP knockdown. Certain groups were treated with (PA) and the fat metabolism, as well as inflammatory and oxidative stress markers were measured. PA accelerated lipid metabolism derangement, inflammatory reaction and oxidative stress in podocytes. Overexpression of H-FABP enhanced the PA-induced disequilibrium in podocytes. The mRNA and protein expression levels of acyl-coenzyme A oxidase 3 and monocyte chemotactic protein 1, and the protein expression levels of 8-hydroxy-2\'-deoxyguanosine and 4-hydroxynonenal were upregulated in the H-FABP overexpression group, while the mRNA and protein expression of peroxisome proliferator activated receptor α was downregulated. Knockdown of H-FABP inhibited the PA-induced injury and lipid metabolism derangement, as well as the inflammatory reaction and oxidative stress in podocytes. These results indicated that overexpression of H-FABP enhances fatty -induced podocyte injury, while H-FABP inhibition may represent a potential therapeutic strategy for the prevention of lipid metabolism-associated podocyte injury.

Keyword: obesity

Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

increases the risk of developing diabetes mellitus. Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in diabetes with . Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II diabetes. Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling pathways was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of diabetes with that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for diabetes with .

Keyword: obesity

Wnt7a promotes wound healing by regulation of angiogenesis and inflammation: Issues on diabetes and .

Diabetic skin heals wounds poorly. Though is the common risk factor of diabetes mellitus, few studies have investigated its effects on wound healing.This study aimed to evaluate the morphology and possible mechanism of human umbilical vein endothelial cells (HUVEC-C) in response to different levels of glucose and , and explore the role of Wnt7a in wound healing.The functional changes of HUVEC-C and mRNA expression of Wnt signaling were determined by analyzing cell viability, migration, tube formation and rt-PCR in gradients of glucose and . Recombinant Wnt7a protein was injected around wounds made on streptozotocin (STZ) -induced diabetic rats with (HF) or without (DM) high-fat diet. Angiogenesis and inflammatory statement were mainly analyzed by immunohistochemistry, ELISA, cytometry and Western blotting.The expression of Wnt7a significantly decreased in high Glc/PA cultured cells or DM and HF wounded rats. Impaired wound healing was also observed in DM and HF groups. The healing rate significantly accelerated after localized injection with Wnt7a at d10. Moreover, the expression of CD31, eNOS phosphorylation and NO were increased; the reduction of local neutrophils influx, ICAM-1 and IL-6/8 expression levels were obvious especially in diabetic with rats at d10 after Wnt7a treatment.This study indicates the potential role of Wnt7a, which is beneficial for regeneration of damaged vessels, moderation of inflammatory statement in diabetic wound healing with or without , thus demonstrating its possible utility as a topical administration to promote healing rate.Copyright © 2018. Published by Elsevier B.V.

Keyword: obesity

Consumption of a high fat diet promotes protein O-GlcNAcylation in mouse retina via NR4A1-dependent GFAT2 expression.

The incidence of type 2 diabetes, the most common cause of diabetic retinopathy (DR), is rapidly on the rise in developed countries due to overconsumption of calorie rich diets. Using an animal model of diet-induced /pre-diabetes, we evaluated the impact of a diet high in saturated fat (HFD) on O-GlcNAcylation of retinal proteins, as dysregulated O-GlcNAcylation contributes to diabetic complications and evidence supports a role in DR. Protein O-GlcNAcylation was increased in the retina of mice fed a HFD as compared to littermates receiving control chow. Similarly, O-GlcNAcylation was elevated in retinal Müller cells in culture exposed to the saturated fatty palmitate or the ceramide analog Cer6. One potential mechanism responsible for elevated O-GlcNAcylation is increased flux through the hexosamine biosynthetic pathway (HBP). Indeed, inhibition of the pathway\'s rate-limiting enzyme glutamine-fructose-6-phosphate amidotransferase (GFAT) prevented Cer6-induced O-GlcNAcylation. Importantly, expression of the mRNA encoding GFAT2, but not GFAT1 was elevated in both the retina of mice fed a HFD and in retinal cells in culture exposed to palmitate or Cer6. Notably, expression of nuclear receptor subfamily 4 group A member 1 (NR4A1) was increased in the retina of mice fed a HFD and NR4A1 expression was sufficient to promote GFAT2 mRNA expression and O-GlcNAcylation in retinal cells in culture. Whereas palmitate or Cer6 addition to culture medium enhanced NR4A1 and GFAT2 expression, chemical inhibition of NR4A1 transactivation repressed Cer6-induced GFAT2 mRNA expression. Overall, the results support a model wherein HFD increases retinal protein O-GlcNAcylation by promoting NR4A1-dependent GFAT2 expression.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

Caveolin-1 prevents palmitate-induced NF-κB signaling by inhibiting GPRC5B-phosphorylation.

Tyrosine phosphorylation of GPRC5B and phosphorylation-dependent recruitment of Fyn through the SH2 domain have been implicated in NF-κB activation and -linked adipose inflammation. GPRC5B tightly associates with caveolin-1 (Cav1); however, the role of this interaction remains elusive. Here, we report that Cav1 reduces GPRC5B-mediated NF-κB signaling by blocking GPRC5B-phosphorylation. We demonstrate highly abundant tyrosine phosphorylation of GPRC5B is observed in Neuro2a cells lacking endogenous Cav1 expression. Reversely, exogenous expression of Cav1 in these cells inhibits GPRC5B-phosphorylation. Although GPRC5B lacks conventional caveolin-binding motif, cytoplasmic tail of GPRC5B directly interacts with the C-terminal domain of Cav1. The vacant scaffolding domain of Cav1 in the protein complex suggests a potential mechanism for blocking GPRC5B-phosphorylation by Cav1, because Fyn loses the activity by binding with Cav1-scaffolding domain. Enhanced GPRC5B-mediated NF-κB signaling in Cav1-deficient cells were observed under palmitate-induced metabolic stress. These results support Cav1 functions as a negative modulator for GPRC5B action.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Ablation of TMEM126B protects against heart injury via improving mitochondrial function in high fat diet (HFD)-induced mice.

The mitochondrial dysfunction in the pathogenesis of myocardial damage associated with high fat diet (HFD)-induced remains largely unknown. Transmembrane protein 126B (TMEM126B), as a complex I assembly factor, plays a key role in regulating mitochondrial function. In the present study, the effects of TMEM126B on mitochondrial function were investigated using genetic knockout approach in HFD-induced mouse models with . We found that TMEM126B was significantly increased in HFD-treated cardiac samples. Genetic ablation of TMEM126B alleviated HFD-mediated metabolic disorder and heart injury. TEM results suggested that cardiac mitochondrial integrity was improved in TMEM126B knockout mice compared with the wild type (WT) mice after HFD challenge. Additionally, the mitochondrial dysfunction induced by HFD was alleviated in mice with TMEM126B knockout, as evidenced by the decreased protein expression levels of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and increased expression of mitofusin-1 (MFN1). The mitochondrial impairments were further confirmed in (PA)-incubated cardiomyocytes, as evidenced by the down-regulated membrane potential and ATP levels, and by the up-regulated mitochondrial reactive oxygen species (ROS) production and DNA damage, which were significantly reversed by TMEM126B knockdown in\xa0vitro. Finally, TMEM126B ablation suppressed mitochondrial-dependent apoptotic death in the hearts of HFD mice. Therefore, TMEM126B led to mitochondrial impairments, contributing to the pathogenesis of HFD-induced cardiac injury, and blockage of TMEM126B could inhibit mitochondrial dysfunction, paving the road to new therapeutic modalities for the prevention of -associated heart injury.Copyright © 2019. Published by Elsevier Inc.

Keyword: obesity

Trans-Fats Inhibit Autophagy Induced by Saturated Fatty Acids.

Depending on the length of their carbon backbone and their saturation status, natural fatty acids have rather distinct biological effects. Thus, longevity of model organisms is increased by extra supply of the most abundant natural cis-unsaturated fatty , oleic , but not by that of the most abundant saturated fatty , . Here, we systematically compared the capacity of different saturated, cis-unsaturated and alien (industrial or ruminant) trans-unsaturated fatty acids to provoke cellular stress in vitro, on cultured human cells expressing a battery of distinct biosensors that detect signs of autophagy, Golgi stress and the unfolded protein response. In contrast to cis-unsaturated fatty acids, trans-unsaturated fatty acids failed to stimulate signs of autophagy including the formation of GFP-LC3B-positive puncta, production of phosphatidylinositol-3-phosphate, and activation of the transcription factor TFEB. When combined effects were assessed, several trans-unsaturated fatty acids including elaidic (the trans-isomer of oleate), linoelaidic , trans-vaccenic and palmitelaidic , were highly efficient in suppressing autophagy and endoplasmic reticulum stress induced by , but not by oleic . Elaidic also inhibited autophagy induction by in vivo, in mouse livers and hearts. We conclude that the well-established, though mechanistically enigmatic toxicity of trans-unsaturated fatty acids may reside in their capacity to abolish cytoprotective stress responses induced by saturated fatty acids.Copyright © 2018 German Center for Neurodegenerative Diseases (DZNE). Published by Elsevier B.V. All rights reserved.

Keyword: obesity

Stimulating effect of palmitate and insulin on cell migration and proliferation in PNT1A and PC3 prostate cells: Counteracting role of metformin.

A potential association between and prostate cancer has been proposed. Metformin, an antidiabetes drug, has antiproliferative effects being proposed for cancer treatment. However, under intense proliferative stimulation conditions such as those found in , its efficacy is still uncertain. Thus, we analyzed the effects of saturated fatty and/or insulin under high concentrations, with or without metformin, on the proliferation and migration of prostate cells.Human prostate epithelial cell lines non-tumor (PNT1A) and tumor (PC3) were treated with control media (DMEM, C), palmitate (100\u2009µM, HF), and/or insulin (50\u2009µU, HI) with or without metformin (100\u2009µM) for 24 or 48\u2009h.Both PNT1A and PC3 cells had greater proliferation when treated with HF, while HI treatment stimulated only PNT1A. Metformin inhibited cell proliferation caused by HF in both cell lines, but it did not block the proliferative action of HI in PNT1A cells. PNT1A increased cell migration after all treatments, while only HF influenced PC3; metformin inhibited the migration stimulated by all obese microenvironments. Both HF and HI treatments in PNT1A and HF treatment in PC3 augmented vimentin expression, resulting in a higher epithelial-mesenchymal transition (which, in turn, could influence cell migration). Metformin inhibited vimentin expression in both normal and tumor cells. Although HF treatment had increased AMPK activation, it also increased the levels of activated ERK1/2, which could be responsible for high cell proliferation in both cell lines. In contrast, HI decreased AMPK activation in both cell lines, whereas it increased ERK1/2 levels in PNT1A and decreased them in PC3 (reflecting greater cell proliferation only in non-tumor cells). Metformin maintained high activation of AMPK and decreased ERK1/2 levels after HF in both cell lines and only after HI in PNT1A, which was able to decrease the cell proliferation triggered by these treatments.Higher concentrations of palmitate on PC3 cells and palmitate and insulin on PNT1A cells stimulate cellular activities that could favor cancer progression. Metformin inhibited most of these stimuli, showing the efficacy of this drug for cancer adjuvant therapy in obese patients (a group at increased risk for the development of prostrate cancer).© 2018 Wiley Periodicals, Inc.

Keyword: obesity

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and . However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: obesity

Nutraceutical Potential of in Metabolic Syndrome.

L. is a well-known fruit worldwide, and its highest production occurs in tropical and subtropical regions. The pulp contains vitamins A, C, and E, B complex vitamins, such as pantothenic and folate, and minerals, such as magnesium and potassium, as well as food fibers. Phenolic compounds, such as benzyl isothiocyanate, glucosinolates, tocopherols (α and δ), β-cryptoxanthin, β-carotene and carotenoids, are found in the seeds. The oil extracted from the seed principally presents oleic fatty followed by , linoleic and stearic acids, whereas the leaves have high contents of food fibers and polyphenolic compounds, flavonoids, saponins, pro-anthocyanins, tocopherol, and benzyl isothiocyanate. Studies demonstrated that the nutrients present in its composition have beneficial effects on the cardiovascular system, protecting it against cardiovascular illnesses and preventing harm caused by free radicals. It has also been reported that it aids in the treatment of diabetes mellitus and in the reduction of cholesterol levels. Thus, both the pulp and the other parts of the plant (leaves and seeds) present antioxidant, anti-hypertensive, hypoglycemic, and hypolipidemic actions, which, in turn, can contribute to the prevention and treatment of and associated metabolic disorders.

Keyword: obesity

An anti-inflammatory chalcone derivative prevents heart and kidney from hyperlipidemia-induced injuries by attenuating inflammation.

is a growing pandemic in both developed and developing countries. Lipid overload in generates a chronic, low-grade inflammation state. Increased inflammation in heart and renal tissues has been shown to promote the progression of heart and renal damage in . Previously, we found that a novel chalcone derivative, L6H21, inhibited lipopolysaccharide-induced inflammatory response. In the present study, we investigated the effects of L6H21 on inflammatory responses in culture and in animal models of lipid overload. We utilized (PA) challenging in mouse peritoneal macrophages and apolipoprotein E knockout (ApoE) mice fed a high fat diet (HFD) to study whether L6H21 mitigates the inflammatory response. Our studies show that L6H21 significantly reduced PA-induced expression of inflammatory cytokines in macrophages by inhibiting mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NFκB) signaling pathways. L6H21 also reduced fibrosis in the kidney and heart tissues, and indices of inflammatory response in the ApoE mice fed a HFD. These effects in vivo were also associated with inhibition of MAPK and NFκB signaling by L6H21. These findings strongly suggest that L6H21 may be a potential agent for high fat diet-induced injuries in heart and kidney.Copyright © 2017. Published by Elsevier Inc.

Keyword: obesity

Phthalate exposure and childhood overweight and : Urinary metabolomic evidence.

Metabolomics may unravel global metabolic changes in response to environmental exposures and identify important biological pathways involved in the pathophysiology of childhood . Phthalate has been considered an obesogen and contributing to overweight and in children. The purpose of this study is to evaluate changes in urine metabolites in response to the environmental phthalate exposure among overweight or obese children, and to investigate the metabolic mechanisms involved in the obesogenic effect of phthalate on children at puberty.Within the national Puberty Timing and Health Effects in Chinese Children (PTHEC) study, 69 overweight/obese children and 80 normal weight children were selected into the current study according to their puberty timing and WGOC (The Working Group for in China) references. Urinary concentrations of six phthalate monoesters (MMP, MEP, MnBP, MEHP, MEOHP and MEHHP) were measured using API 2000 electrospray triple quadrupole mass spectrometer (ESIMS/MS). Metabolomic profiling of spot urine was performed using gas chromatography-mass spectrometry. Differentially expressed urinary metabolites associated with phthalate monoesters exposure were examined using orthogonal partial least square-discriminant analysis and multiple linear regression models. In addition, the candidate metabolites were regressed to indices with multiple linear regression models and logistic regression models in all subjects.Compared with normal weight children, higher levels of MnBP were detected in urinary samples of children with overweight and . After adjusting for confounders including chronological age, gender, puberty onset, daily energy intake and physical activity and socio-economic level, positive association remained between urinary MnBP concentration and childhood overweight/ [OR\u202f=\u202f1.586, 95% CI:1.043,2.412]. We observed elevated MnBP concentration was significantly correlated with increased levels of monostearin, 1-monopalmitin, stearic , itaconic , glycerol 3-phosphate, 5-methoxytryptamine, kyotorphin, 1-methylhydantoin, d-alanyl-d-alanine, pyrrole-2-carboxylic , 3,4-Dihydroxyphenylglycol, and butyraldehyde. Meanwhile, increased MnBP concentration was also significantly correlated with decreased levels of lactate, glucose 6-phosphate, d-fructose 6-phosphate, , 4-acetamidobutyric , l-glutamic , n-acetyl-l-phenylalanine, iminodiacetic , hydroxyproline, pipecolinic , l-ornithine, n-acetyl-l-glutamic , guanosine, cytosin, and (s)-mandelic in the normal weight subjects. The observations indicated that MnBP exposure was related to global urine metabolic abnormalities characterized by disrupting arginine and proline metabolism and increasing oxidative stress and fatty reesterification. Among the metabolic markers related to MnBP exposure, 1-methylhydantoin, pyrrole-2-carboxylic and monostearin were found to be positively correlated with indices, while hydroxyproline, l-ornithine, and lactate were negatively associated with overweight/ in children.Our results suggested that the disrupted arginine and proline metabolism associated with phthalate exposure might contribute to the development of overweight and in school-age children, providing insights into the pathophysiological changes and molecular mechanisms involved in childhood .Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: obesity

Dietary saturated fatty type impacts -induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with , insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces -induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low fat diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic lipid accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and diabetes, and (3) dietary SFA type impacts LCPUFA metabolism.Published by Elsevier Inc.

Keyword: obesity

Pediatric Age Palm Oil Consumption.

Palm oil is widely used in the food industry for its chemical/physical properties, low cost and wide availability. Its widespread use has provoked an intense debate about whether it is a potential danger to human health. In a careful review of the scientific literature, we focused on nutritional characteristics and health effects of the use of palm oil with regards to children, seeking to determine whether there is evidence that justifies fears about the health effects of palm oil. Our review showed that palm oil represents a significant source of saturated fatty acids, to which scientific evidence attributes negative health effects when used in excess, especially with regards to cardiovascular diseases. However, to date, there is no evidence about the harmful effects of palm oil on the health of children. Nevertheless, palm oil has possible ill health effects linked to its composition of fatty acids: its consumption is not correlated to risk factors for cardiovascular diseases in young people with a normal weight and cholesterol level; the elderly and patients with dyslipidaemia or previous cardiovascular events or hypertension are at a greater risk. Therefore, the matter is not palm oil itself but the fatty--rich food group to which it belongs. The most important thing is to consume no more than 10% of saturated fatty acids, regardless of their origin and regardless of one\'s age. Correct information based on a careful analysis of the scientific evidence, rather than a focus on a singular presumed culprit substance, should encourage better lifestyles.

Keyword: obesity

The impact of in the cardiac lipidome and its consequences in the cardiac damage observed in obese rats.

To explore the impact of on the cardiac lipid profile in rats with diet-induced , as well as to evaluate whether or not the specific changes in lipid species are associated with cardiac fibrosis.Male Wistar rats were fed either a high-fat diet (HFD, 35% fat) or standard diet (3.5% fat) for 6 weeks. Cardiac lipids were analyzed using by liquid chromatography-tandem mass spectrometry.HFD rats showed cardiac fibrosis and enhanced levels of cardiac superoxide anion (O), HOMA index, adiposity, and plasma leptin, as well as a reduction in those of cardiac glucose transporter (GLUT 4), compared with control animals. Cardiac lipid profile analysis showed a significant increase in triglycerides, especially those enriched with , stearic, and arachidonic . An increase in levels of diacylglycerol (DAG) was also observed. No changes in cardiac levels of diacyl phosphatidylcholine, or even a reduction in total levels of diacyl phosphatidylethanolamine, diacyl phosphatidylinositol, and sphingomyelins (SM) was observed in HFD, as compared with control animals. After adjustment for other variables (oxidative stress, HOMA, cardiac hypertrophy), total levels of DAG were independent predictors of cardiac fibrosis while the levels of total SM were independent predictors of the cardiac levels of GLUT 4.These data suggest that has a significant impact on cardiac lipid composition, although it does not modulate the different species in a similar manner. Nonetheless, these changes are likely to participate in the cardiac damage in the context of , since total DAG levels can facilitate the development of cardiac fibrosis, and SM levels predict GLUT4 levels.Copyright © 2017 Sociedad Española de Arteriosclerosis. Publicado por Elsevier España, S.L.U. All rights reserved.

Keyword: obesity

is a novel hypothalamic gene upregulated by a high-fat diet and leptin in mice.

Energy homeostasis is regulated by the hypothalamus but fails when animals are fed a high-fat diet (HFD), and leptin insensitivity and develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated fat, oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic inflammation.

Keyword: obesity

damages gut epithelium integrity and initiates inflammatory cytokine production.

The mechanisms leading to the low-grade inflammation observed during are not fully understood. Seeking the initiating events, we tested the hypothesis that the intestine could be damaged by repeated lipid supply and therefore participate in inflammation. In mice, 1-5 palm oil gavages increased intestinal permeability via decreased expression and mislocalization of junctional proteins at the cell-cell contacts; altered the intestinal bacterial species by decreasing the abundance of Akkermansia muciniphila, segmented filamentous bacteria, and Clostridium leptum; and increased inflammatory cytokine expression. This was further studied in human intestinal epithelial Caco-2/TC7 cells using the two main components of palm oil, i.e., and oleic . Saturated impaired paracellular permeability and junctional protein localization, and induced inflammatory cytokine expression in the cells, but unsaturated oleic did not. Inhibiting de novo ceramide synthesis prevented part of these effects. Altogether, our data show that short exposure to palm oil or induces intestinal dysfunctions targeting barrier integrity and inflammation. Excessive palm oil consumption could be an early player in the gut alterations observed in metabolic diseases.Copyright © 2019. Published by Elsevier B.V.

Keyword: obesity

Euodia daniellii Hemsl. (Bee-Bee Tree) Oil Attenuates Palmitate-Induced Lipid Accumulation and Apoptosis in Hepatocytes.

Hepatic lipid accumulation and apoptosis is elevated in patients with non-alcoholic steatohepatitis and is closely associated with severity. Saturated fatty palmitate stimulates lipid accumulation and apoptosis in hepatocytes. In the present study, we examined bee-bee tree oil (BO)-mediated protective effects on palmitate-induced lipid accumulation and apoptosis in mouse primary hepatocytes. Cells were cultured in a control media or the same media containing 150 or 300 µmol/L of albumin-bound palmitate for 24 h. BO concentrations used were 0, 0.1, 0.2, or 0.5%. Palmitate induced lipid accumulation and mRNA expression of lipogenic genes such as SREBP1c and SCD1. However, BO prevented these changes. Furthermore, palmitate stimulated caspase-3 activity and decreased cell viability in the absence of BO. BO reduced palmitate-induced activation of caspase-3 and cell death in a dose-dependent manner. AMP-activated protein kinase inhibitors abolished the effects of BO. Furthermore, BO suppressed palmitate-induced c-Jun N-terminal kinase (JNK) phosphorylation through the 5' adenosine monophosphate-activated protein kinase (AMPK)-dependent pathway. In conclusion, BO attenuated palmitate-induced hepatic steatosis and apoptosis through AMPK-mediated suppression of JNK signaling. These data suggest that BO is an important determinant of saturated fatty -induced lipid accumulation and apoptosis, and may be an effective therapeutic strategy for treatment of -mediated liver diseases.© 2018 S. Karger AG, Basel.

Keyword: obesity

-related cellular stressors regulate gonadotropin releasing hormone gene expression via c-Fos/AP-1.

is a risk factor for infertility, but mechanisms underlying this risk are unclear. Fertility is regulated by hypothalamic gonadotropin-releasing hormone, encoded by the Gnrh1 gene. Because promotes endoplasmic reticulum (ER) stress, we sought to determine how tunicamycin-induced ER stress affected Gnrh1 gene expression in the mouse hypothalamic cell line GT1-7. Tunicamycin repressed expression of Gnrh1 in a PKC- and JNK-dependent manner, while upregulating expression of a known Gnrh1 repressor, Fos. is associated with increased circulating free fatty acids, and exposure to palmitate promoted ER stress and inflammation. Fos expression increased with palmitate dose, but Gnrh1 expression was upregulated with low-dose palmitate and repressed with high-dose palmitate. Using a small molecule inhibitor, we determined that AP-1 was required for Gnrh1 repression by high-dose palmitate or tunicamycin-induced ER stress. These findings suggest that hypogonadism driven by decreased hypothalamic GnRH may be a component of -related infertility.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: obesity

MicroRNA-155 Mediates -Induced Renal Inflammation and Dysfunction.

Chronic inflammation is a major contributor to -related renal damage. Recent studies have demonstrated that microRNA (miR)-155 is closely associated with hyperglycemia-induced nephropathy, but whether renal miR-155 participates in the inflammatory response and development of -related nephropathy is unknown. In present study, we investigated the pathophysiological role of renal miR-155 in (PA)-treated endothelial cell and high-fat-diet (HFD)-fed mouse models by specific miR-155 sponge. Mice fed with HFD exhibited higher levels of renal miR-155, which positively correlated with urine microalbumin and blood urea nitrogen. In vitro study, mouse renal vascular endothelial cells stimulated with PA also showed higher miR-155 levels, accompanied with increased inflammatory response. Suppression of renal miR-155 effectively attenuated HFD-induced renal structural damages and dysfunction. MiR-155 sponge treatment also significantly decreased NF-κB signaling and downstream gene expression in vitro and in vivo. The -increased macrophage infiltration and lipotoxicity was decreased in mouse kidney after miR-155 sponge treatment. Mechanistically, miR-155 directly targeted 3\'-UTR of SHIP1/INPP5D and suppressed its expression in vitro and in vivo, whereas silence of SHIP1/INPP5D abolished the renal protective benefits of miR-155 sponge in obese mice. Taken together, present findings for the first time provided evidence for the potential role of miR-155 in -related nephropathy and clarified that SHIP1/NF-κB signaling was a potential molecular mechanism.

Keyword: obesity

In vivo kinetic study of materno-fetal fatty transfer in obese and normal weight pregnant women.

Placental structure and function can be modified as a result of maternal affecting materno-fetal fatty acids (FA) transport. We report for the first time, in humans and in vivo, the kinetics of placental FA transfer in normo-weight and in normolipemic obese pregnant women using stable isotopes. The administration of different tracer FA with similar behaviour to the mother at different time points allows the collection of kinetic information on materno-fetal transfer of FA despite only one sample of placenta and cord can be collected per subject. Computational modelling showed a good fit to the data when considering all maternal plasma lipid classes but not when based only on non-esterified FA. The novel approach using multiple tracer FA administration combined with computational modelling shows a consistent time course of placental tracer FA and predicted total FA accumulation.We analyse for the first time the in vivo materno-fetal kinetic transfer of fatty acids (FA) labelled with stable isotopes in control and obese (OB) pregnant women. Labelled FA with a similar metabolism (stearic : C-SA; : C-PA; oleic : C-OA) were orally administered at -4\xa0h, -8\xa0h and -12\xa0h, respectively prior to elective caesarean section to 10 pregnant women with a body mass index >30 (OB) and 10 with a body mass index in the range 20-25 (NW). Placenta, venous and arterial cord blood were collected obtaining a wide range of FA enrichments. A combined experimental and computational modelling analysis was applied. FA fractional synthesis rate (FSR) in placenta was 11-12%\xa0h . No differences were observed between NW and normo-lipidemic OB. It was not possible to estimate FA FSR in cord blood with this oral bolus dose approach. Computational modelling demonstrated a good fit to the data when all maternal plasma lipid classes were included but not with modelling based only on the non-esterified FA fraction. The estimated materno-fetal C-FA transfer was ∼1%. In conclusion, our approach using multiple C-FA tracers allowed us to estimated FSR in placental/maternal plasma but not in fetal/maternal compartments. Computational modelling showed a consistent time course of placental C-FA transfer and predicted total fetal FA accumulation during the experiment. We conclude that, in addition to non-esterified FA fraction in the maternal circulation, maternal plasma very low-density lipoprotein and other lipoproteins are important contributors to placental FA transfer to the fetus.© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.

Keyword: obesity

Synthesis and biological evaluations of marine oxohexadecenoic acids: PPARα/γ dual agonism and anti-diabetic target gene effects.

and associated disorders such as metabolic syndrome and type 2 diabetes (T2D) have reached epidemic proportions. Several natural products have been reported as Peroxisome Proliferator-Activated Receptor (PPAR) agonists, functioning as lead compounds towards developing new anti-diabetic drugs due to adverse side effects of existing PPAR drugs. We recently isolated and identified (7E)-9-oxohexadec-7-enoic (1) and (10E)-9-oxohexadec-10-enoic (2) from the marine algae Chaetoceros karianus. Herein we report the total synthesis, pharmacological characterization, and biological evaluations of these naturally occurring oxo-fatty acids (oFAs). The syntheses of 1 and 2 afforded sufficient material for extensive biological evaluations. Both oFAs show an appreciable dose-dependent activation of PPARα and -γ, with EC values in the micromolar range, and an ability to regulate important PPAR target genes in hepatocytes and adipocytes. Moreover, both 1 and 2 are able to drive adipogenesis when evaluated in the Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocyte cell model, but with lowered expression of adipocyte markers and reduced lipid accumulation compared to the drug rosiglitazone. This seems to be caused by a transient upregulation of PPARγ and C/EBPα expression. Importantly, whole transcriptome analysis shows that both compounds induce anti-diabetic gene programs in adipocytes by upregulating insulin-sensitizing adipokines and repressing pro-inflammatory cytokines.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: obesity

High-fat diet consumption reduces hepatic folate transporter expression via nuclear respiratory factor-1.

Folate is an essential micronutrient for biological function. The liver, a primary organ for folate metabolism and storage, plays an important role in folate homeostasis. Proton-coupled folate transporter (PCFT) and reduced folate carrier (RFC) are the major folate transporters responsible for folate uptake at basolateral membrane of hepatocytes. Low serum folate levels are frequently associated with . We investigated the mechanism that regulated folate status in a mouse model with diet-induced . Mice (C57BL/6J) were fed a high-fat diet (60% kcal fat) for 8\xa0weeks. Mice displayed increased hepatic lipid accumulation and decreased folate levels in the liver and serum compared to mice fed a normal chow diet (10% kcal fat). High-fat diet-fed mice had low expression of PCFT and RFC and decreased nuclear respiratory factor-1 (NRF-1)/DNA-binding activity. Treatment with NRF-1 siRNA or reduced folate transporter expression in hepatocytes. Inhibition of NRF-1 mediated folate transporter expression significantly reduced intracellular folate levels. These results suggest that chronic consumption of high-fat diets impairs folate transporter expression via NRF-1-dependent mechanism, leading to reduced hepatic folate storage. Understanding the regulation of folate homeostasis in may have an important implication in current guideline of folate intake. KEY MESSAGES: Serum and liver folate levels are decreased in diet-induced obese mice. Chronic high-fat diet consumption impairs expression of hepatic PCFT and RFC. NRF-1 regulates hepatic folate transporters expression and folate levels.

Keyword: obesity

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the liver was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

Keyword: obesity

The effect of on inflammatory response in macrophages: an overview of molecular mechanisms.

is a saturated fatty whose blood concentration is elevated in obese patients. This causes inflammatory responses, where toll-like receptors (TLR), TLR2 and TLR4, play an important role. Nevertheless, is not only a TLR agonist. In the cell, this fatty is converted into phospholipids, diacylglycerol and ceramides. They trigger the activation of various signaling pathways that are common for LPS-mediated TLR4 activation. In particular, metabolic products of affect the activation of various PKCs, ER stress and cause an increase in ROS generation. Thanks to this, also strengthens the TLR4-induced signaling. In this review, we discuss the mechanisms of inflammatory response induced by . In particular, we focus on describing its effect on ER stress and IRE1α, and the mechanisms of NF-κB activation. We also present the mechanisms of inflammasome NLRP3 activation and the effect of on enhanced inflammatory response by increasing the expression of FABP4/aP2. Finally, we focus on the consequences of inflammatory responses, in particular, the effect of TNF-α, IL-1β and IL-6 on insulin resistance. Due to the high importance of macrophages and the production of proinflammatory cytokines by them, this work mainly focuses on these cells.

Keyword: obesity

Attenuation of Free Fatty -Induced Muscle Insulin Resistance by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in , impair muscle insulin action leading to insulin resistance and Type 2 diabetes mellitus. Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties.

Keyword: obesity

Unsaturated Fatty Acids Increase the Expression of Hepassocin through a Signal Transducer and Activator of Transcription 3-Dependent Pathway in HepG2 Cells.

Hepassocin (HPS) is a hepatokine that regulates hepatocyte proliferation. It is known that HPS plays an important role in the development of nonalcoholic fatty liver diseases (NAFLD). Fatty acids, such as oleic (OLA), exhibit the ability to activate the signal transducer and activator of transcription-3 (STAT3), and the binding site of STAT3 is found in the promoter region of HPS. However, the regulation of HPS by fatty acids is still obscure. To clarify the regulation of HPS, we detected the expression of HPS by western blots. In addition, a hepatic steatosis cell culture model was established by treatment of different fatty acids, including linoleic (LNA), oleic , , and stearic . The intracellular lipid accumulation was confirmed by oil red O staining. Blocking of STAT3 activity was achieved by the pretreatment of the STAT3 inhibitor, stattic. We found that activation of STAT3 by interleukin-6 (IL-6) was mediated in the regulation of HPS expression. Treatment of unsaturated fatty acids significantly induced intracellular lipid accumulation in HepG2 cells. Moreover, the expressions of HPS were increased in unsaturated fatty -treated HepG2 cells, as compared with saturated fatty -treated groups. Also, the expression of HPS induced by OLA was blocked by the inhibition of STAT3 activity. Furthermore, we found that deletion of HPS by small interfering ribonucleic transfection decreased the protective effect of OLA on cell viability. Taken together, we provided evidence that STAT3 plays an important role in the regulation of OLA-induced HPS expression and the increased HPS may further participate in the development of NAFLD. In addition, the increase of HPS might be involved in the protective effect of OLA on cell viability.© 2018 AOCS.

Keyword: obesity

Alternation of plasma fatty acids composition and desaturase activities in children with liver steatosis.

The aim of this study was to investigate changes in plasma fatty acids proportions and estimated desaturase activities for variable grading of liver steatosis in children.In total, 111 schoolchildren (aged 8-18 years) were included in the analysis from March 2015 to August 2016. Anthropometric evaluation, liver ultrasound examination and scoring for nonalcoholic fatty liver disease (NAFLD score = 0-6), and biochemical and plasma fatty acids analysis were performed. We compared the composition ratio of fatty acids between children with high-grade liver steatosis (NAFLD score = 4-6), low-grade liver steatosis (NAFLD score = 1-3), and healthy controls (NAFLD score = 0). In addition, correlation coefficients (r) between NAFLD score, metabolic variables, and estimated activity of desaturase indices (stearoyl-coenzyme A desaturase-1 (SCD1), delta-5 and delta-6 desaturase) were calculated.Compared with healthy controls, children with liver steatosis showed a higher proportion of monounsaturated fatty acids (21.16 ± 2.81% vs. 19.68 ± 2.71%, p = 0.024). In addition, children with high- grade liver steatosis exhibited higher proportions of (C16:0), palmitoleic (C16:1n-7), dihomo-γ-linolenic (C20:3n-6), adrenic (C22:4n-6), and docosapentaenoic (C22:5n-6); and lower proportions of eicosapentaenoic (C20:5n-3) (P< 0.05). In all subjects, the NAFLD score was positively correlated with body mass index (BMI) (kg/m2) (r = 0.696), homeostasis model of assessment ratio-index (HOMA-IR) (r = 0.510), SCD1(16) (r = 0.273), and the delta-6 index (r = 0.494); and inversely associated with the delta-5 index (r = -0.443).Our current data suggested that children with liver steatosis was highly associated with , and insulin resistance. In addition, increased endogenous lipogenesis through altered desaturase activity may contribute to the progression of liver steatosis in children.

Keyword: obesity

Degradation of splicing factor SRSF3 contributes to progressive liver disease.

Serine rich splicing factor 3 (SRSF3) plays a critical role in liver function and its loss promotes chronic liver damage and regeneration. As a consequence, genetic deletion of SRSF3 in hepatocytes caused progressive liver disease and ultimately led to hepatocellular carcinoma. Here we show that SRSF3 is decreased in human liver samples with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or cirrhosis that was associated with alterations in RNA splicing of known SRSF3 target genes. Hepatic SRSF3 expression was similarly decreased and RNA splicing dysregulated in mouse models of NAFLD and NASH. We showed that -induced oxidative stress caused conjugation of the ubiquitin like NEDD8 protein to SRSF3 and proteasome mediated degradation. SRSF3 was selectively neddylated at lysine11 and mutation of this residue (SRSF3-K11R) was sufficient to prevent both SRSF3 degradation and alterations in RNA splicing. Finally prevention of SRSF3 degradation in vivo partially protected mice from hepatic steatosis, fibrosis and inflammation. These results highlight a neddylation-dependent mechanism regulating gene expression in the liver that is disrupted in early metabolic liver disease and may contribute to the progression to NASH, cirrhosis and ultimately hepatocellular carcinoma.

Keyword: obesity

Effect of weight loss on circulating fatty profiles in overweight subjects with high visceral fat area: a 12-week randomized controlled trial.

Significant associations between visceral fat and alterations in plasma fatty acids have been identified in overweight individuals. However, there are scant data regarding the relationships of the visceral fat area (VFA) with the plasma fatty profiles and desaturase activities following weight loss. We investigated the effect of weight loss with mild calorie restriction on the circulating fatty profiles and desaturase activities in nondiabetic overweight subjects with high VFA.Eighty overweight subjects with high VFA (L4 VFA ≥100\xa0cm) were randomized into the 12-week mild-calorie-restriction (300\xa0kcal/day) or control groups.Comparison of the percent of body weight changes between groups revealed that the weight-loss group had greater reductions in body weight. The VFA decreased by 17.7\xa0cm from baseline in the weight-loss group\xa0(P < 0.001). At follow-up, the weight-loss group showed greater reductions in serum triglycerides, insulin, and HOMA-IR than the control group. Significantly greater reductions in total saturated fatty acids, , stearic , total monounsaturated fatty acids, palmitoleic , oleic , eicosadienoic , and dihomo-γ-linolenic levels were detected in the weight-loss group compared with the control group after adjusting for baseline values. Following weight loss, C16 Δ9-desaturase activity was significantly decreased and Δ5-desaturase activity was significantly increased, and the changes were greater in the weight-loss group than in the control group.The results suggest that mild weight loss improves abdominal , overall fatty profiles, and desaturase activities; therefore, mild calorie restriction has potential health benefits related to -related diseases in overweight subjects with high VFA.. Retrospectively registered 11 December 2016.

Keyword: obesity

Role of the mTOR‑FOXO1 pathway in ‑associated renal tubulointerstitial inflammation.

Since is largely responsible for the growing incidence of renal tubulointerstitial inflammation, exploration into the mechanisms of ‑associated tubulointerstitial inflammation is essential. Studies have demonstrated that mammalian target of rapamycin\xa0(mTOR) is a crucial molecule in the pathogenesis of renal inflammation, including regulating the expression of inflammatory factors. The purpose of the present study was to further elucidate the role of mTOR in ‑associated tubulointerstitial inflammation. In the clinical study, obese and healthy subjects were recruited for physical examination, as well as the collection of blood and urine samples. Further study was performed on a high fat diet\xa0(HFD)‑induced obese rat model and a cultured human renal tubular epithelial cell line\xa0(HK‑2). The clinical study demonstrated that the participants with had increased serum lipids, creatinine\xa0(Cr), urinary albumin to creatinine ratio\xa0(UACR) and urinary neutrophil gelatinase‑associated lipocalin\xa0(u‑NGAL). Moreover, the level of urinary monocyte chemoattractant protein‑1\xa0(u‑MCP‑1) was increased in the participants with , and it was positively correlated with free fatty \xa0(FFA), UACR and u‑NGAL. In the in\xa0vivo study, the results indicated that the levels of serum lipids, Cr and blood urea nitrogen\xa0(BUN), as well as 24\xa0h urine protein and u‑NGAL, were significantly increased in the HFD‑fed obese rats. In addition, the infiltration of CD68+ cells into the renal interstitial area and the release of interleukin‑1β\xa0(IL‑1β) was observed in the kidneys of obese rats. Meanwhile, the supernatant from HK‑2 cells treated with stimulated THP‑1 monocyte migration. The upregulation of MCP‑1, phosphorylated forkhead boxO1 (p‑FOXO1), and phosphorylated mTOR (p‑mTOR) was observed in\xa0vivo and in\xa0vitro. However, inhibition of mTOR was able to alleviate the above effects. Overall, these results demonstrated that activated mTOR induced FOXO1 phosphorylation, which mediates renal MCP‑1 release, causes tubulointerstitial inflammation and ultimately leads to pathological renal changes and dysfunction. However, inhibition of mTOR may play a renoprotective role during the progression of ‑associated tubulointerstitial inflammation.

Keyword: obesity

Myeloid differentiation protein 1 protected myocardial function against high-fat stimulation induced pathological remodelling.

Myeloid differentiation 1 (MD-1) is a secreted protein that regulates the immune response of B cell through interacting with radioprotective 105 (RP105). Disrupted immune response may contribute to the development of cardiac diseases, while the roles of MD-1 remain elusive. Our studies aimed to explore the functions and molecular mechanisms of MD-1 in -induced cardiomyopathy. H9C2 myocardial cells were treated with free fatty (FFA) containing and oleic to challenge high-fat stimulation and adenoviruses harbouring human MD-1 coding sequences or shRNA for MD-1 overexpression or knockdown in vitro. MD-1 overexpression or knockdown transgenic mice were generated to assess the effects of MD-1 on high-fat diet (HD) induced cardiomyopathy in vivo. Our results showed that MD-1 was down-regulated in H9C2 cells exposed to FFA stimulation for 48\xa0hours and in mice induced by HD for 20\xa0weeks. Both in vivo and in vitro, silencing of MD-1 accelerated myocardial function injury induced by HD stimulation through increased cardiac hypertrophy and fibrosis, while overexpression of MD-1 alleviated the effects of HD by inhibiting the process of cardiac remodelling. Moreover, the MAPK and NF-κB pathways were overactivated in MD-1 deficient mice and H9C2 cells after high-fat treatment. Inhibition of MAPK and NF-κB pathways played a cardioprotective role against the adverse effects of MD-1 silencing on high-fat stimulation induced pathological remodelling. In conclusion, MD-1 protected myocardial function against high-fat stimulation induced cardiac pathological remodelling through negative regulation for MAPK/NF-κB signalling pathways, providing feasible strategies for cardiomyopathy.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: obesity

Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced .

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti- drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased body and liver weights, insulin, leptin, triglyceride, cholesterol and free fatty plasma levels in a mouse model of diet-induced . Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in energy expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

Keyword: obesity

Ceramide stearic to ratio predicts incident diabetes.

Ceramide lipids have a role in the development of insulin resistance, diabetes and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident diabetes.A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the metabolic syndrome (Prevent Metabolic Syndrome [PrevMetSyn], n\u2009=\u2009371). Diabetes risk score models were developed to estimate the 10\xa0year risk of incident diabetes.Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident diabetes (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and diabetes risk scores significantly decreased in individuals who had 5% or more weight loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident diabetes, and may be modulated by lifestyle intervention.

Keyword: obesity

Histone methyltransferase G9a modulates hepatic insulin signaling via regulating HMGA1.

Hepatic insulin sensitivity is critical for glucose homeostasis, and insulin resistance is a fundamental syndrome found in various metabolic disorders, including and type 2 diabetes. Despite considerable studies on the mechanisms of hepatic insulin resistance, the link between epigenetic regulation and the development of insulin resistance remains elusive. Here, we reported that G9a/EHMT2, a histone methyltransferase, was markedly decreased in the liver of db/db mice and high-fat diet (HFD)-fed mice. In cultured hepatic cells, G9a knockdown resulted in downregulation of insulin receptor, p-AKT and p-GSK3β; while upon upregulation, G9a prevented the - or glucosamine-induced insulin resistance by preserving the normal level of insulin receptor and integrity of insulin signaling. Further mechanistic study suggested that G9a regulated the expression level of high mobility group AT-hook 1 (HMGA1), a key regulator responsible for the transcription of insulin receptor (INSR) gene. Overexpression of HMGA1 normalized the impaired insulin signaling in G9a knockdown hepatic cells. Importantly, in db/db mice, restoring the expression level of G9a not only upregulated HMGA1 level and improved the impaired hepatic insulin signaling, but also alleviated hyperglycemia and hyperinsulinemia. Together, our results revealed a novel role for G9a in modulating insulin signaling, at least in part, depending on its regulatory function on HMGA1.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: obesity

Acute and Repeated Treatment with 5-PAHSA or 9-PAHSA Isomers Does Not Improve Glucose Control in Mice.

Fatty esters of hydroxylated fatty acids (FAHFAs) were discovered as a novel class of endogenous mammalian lipids whose profound effects on metabolism have been shown. In the current study, in\xa0vitro and in\xa0vivo the metabolic effects of two of these FAHFAs, namely -5- (or -9) -hydroxy-stearic (5- or 9-PAHSA, respectively) were profiled. In DIO mice fed with differentially composed low- or high-fat diets, acute and subchronic treatment with 5-PAHSA and 9-PAHSA alone, or in combination, did not significantly improve the deranged metabolic status. Neither racemic 5- or 9-PAHSA, nor the enantiomers were able to: (1) increase basal or insulin-stimulated glucose uptake in\xa0vitro, (2) stimulate GLP-1 release from GLUTag cells, or (3) induce GSIS in rat, mouse, or human islets or in a human pancreatic β cell line. Therefore, our data do not support the\xa0further development of PAHSAs or their derivatives for the control of insulin resistance and hyperglycemia.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Kaempferol alleviates -induced lipid stores, endoplasmic reticulum stress and pancreatic β-cell dysfunction through AMPK/mTOR-mediated lipophagy.

Kaempferol, a natural flavonoid, has the beneficial effects of preserving pancreatic β-cell mass and function, but its action on β-cell lipid metabolism still remains elusive. Recently, autophagy has been reported to play a major role in lipid metabolism in various cell types, but its role in pancreatic β-cell\'s lipid metabolism is rarely reported. Here, we investigated the role of kaempferol-induced autophagy in inhibition of lipid stores, ER stress and β-cell dysfunction in -challenged RIN-5F cells and isolated pancreatic islets. The lipid-lowering effect of kaempferol was determined by Oil Red O staining, triglyceride assay, BODIPY labeling, RT-PCR and immunoblot analysis of PLIN2 (the lipid droplet coat protein) expression. Further, the involvement of AMPK/mTOR-mediated lipophagy was established by pharmacological and genetic inhibitors of autophagy and AMPK. The co-localization studies of lipid droplets with autophagosomes/lysosomes by BODIPY-MDC-LysoTracker co-staining, LC3/BODIPY labeling and LC3/PLIN2 double immunolabeling further strengthened the findings. Kaempferol treatment exhibited decreased lipid stores and increased co-localization of lipid droplets with autophagosomes and lysosomes in -challenged β-cells. Moreover, inhibition of autophagy led to decreased co-localization and increased lipid droplets accumulation. Kaempferol-induced alleviation of ER stress and β-cell dysfunctions was established by immunoblot analysis of CHOP-10 (a key mediator of cell death in response to ER stress) and insulin content/secretion analysis respectively. Together, these findings suggest that kaempferol prevents ectopic lipid accumulation and ER stress, thus restoring β-cell function through AMPK-mediated lipophagy. The current data implies that kaempferol may be a potential therapeutic candidate to prevent -linked diabetic complications.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Improvement of Lipid and Glucose Metabolism by Capsiate in -Treated HepG2 Cells via Activation of the AMPK/SIRT1 Signaling Pathway.

Capsiate, a nonpungent ingredient of CH-19 Sweet, exhibits anti- effects on animals and humans. This study investigated the effects and molecular mechanism of capsiate on lipid and glucose metabolism in PA-treated HepG2 cells. Results showed that compared with the PA-alone group, 100 μM capsiate inhibited lipid accumulation, decreased TG (0.0562 ± 0.0142 vs 0.0381 ± 0.0055 mmol/g of protein; P = 0.024) and TC (0.1087 ± 0.0037 vs 0.0359 ± 0.0059 mmol/g of protein; P = 0.000) levels, and increased the HDL-C level (0.0189 ± 0.0067 vs 0.1050 ± 0.0106 mmol/g of protein; P = 0.000) and glycogen content (0.0065 ± 0.0007 vs 0.0146 ± 0.0008 mg/10 cells; P = 0.000) of PA-treated HepG2 cells; 100 μM capsiate also upregulated the level of CD36 ( P = 0.000), phosphorylation of ACC ( P = 0.034), and expression of CPT1 ( P = 0.013) in PA-treated HepG2 cells, leading to an enhancement of lipid metabolism. Meanwhile, 100 μM capsiate upregulated the levels of GLUT1, GLUT4, GK, and phosphorylation of GS ( P = 0.001, 0.029, 0.000, and 0.045, respectively) and downregulated the PEPCK level ( P = 0.001) to improve glucose metabolism in PA-treated HepG2 cells. Furthermore, the levels of phosphorylation of AMPK and expression of SIRT1 in HepG2 cells were increased by a 100 μM capsiate treatment ( P = 0.001 and 0.000, respectively), while the FGF21 level was decreased ( P = 0.003). Most of these effects were reversed by pretreatment with compound C, a selective AMPK inhibitor. Thus, capsiate might improve lipid and glucose metabolism in HepG2 cells by activating the AMPK/SIRT1 signaling pathway.

Keyword: obesity

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, , and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe liver inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: obesity

NF‑κB‑miR15a‑bFGF/VEGFA axis contributes to the impaired angiogenic capacity of BM‑MSCs in high fat diet‑fed mice.

Potent paracrine properties, such as secretion of angiogenic cytokines and growth factors, have been considered essential for the function of mesenchymal stem cells (MSCs) in tissue regeneration and repair. The present study determined that bone marrow‑derived mesenchymal stem cells from mice fed a high fat diet (HFD) had reduced pro‑angiogenic capacity, as evident from the reduced expression of vascular endothelial growth factor A (VEGFA) and basic fibroblast growth factor (bFGF); therefore, a reduced number of branches was induced in the angiogenesis assay. Additionally, the present study determined that miR‑15a, a putative microRNA targeting both VEGFA and bFGF, may simultaneously downregulate bFGF and VEGFA expression levels through the 3\'‑untranslated region. Inhibition of miR‑15a using an antagonist restored the expression of VEGFA and bFGF under fatty treatment and thus the angiogenic capacity. Furthermore, the HFD and fatty acids treatments transcriptionally activated the expression of miR‑15a via nuclear factor‑κB. In conclusion, the findings of the present study revealed that inhibition of miR‑15a may restore the therapeutic efficacy of mesenchymal stem cells in patients suffering from .

Keyword: obesity

Macrophage alternative activation confers protection against lipotoxicity-induced cell death.

Alternative activation (M2) of adipose tissue resident macrophage (ATM) inhibits -induced metabolic inflammation. The underlying mechanisms remain unclear. Recent studies have shown that dysregulated lipid homeostasis caused by increased lipolysis in white adipose tissue (WAT) in the obese state is a trigger of inflammatory responses. We investigated the role of M2 macrophages in lipotoxicity-induced inflammation.We used microarray experiments to profile macrophage gene expression regulated by two M2 inducers, interleukin-4 (Il-4), and peroxisome proliferator-activated receptor delta/gamma (Pparδ/Pparγ) agonists. Functional validation studies were performed in bone marrow-derived macrophages and mice deprived of the signal transducer and activator of transcription 6 gene (Stat6; downstream effector of Il-4) or Pparδ/Pparγ genes (downstream effectors of Stat6). (PA) and β-adrenergic agonist were employed to induce macrophage lipid loading in\xa0vitro and in\xa0vivo, respectively.Profiling of genes regulated by Il-4 or Pparδ/Pparγ agonists reveals that alternative activation promotes the cell survival program, while inhibiting that of inflammation-related cell death. Deletion of Stat6 or Pparδ/Pparγ increases the susceptibility of macrophages to PA-induced cell death. NLR family pyrin domain containing 3 (Nlrp3) inflammasome activation by PA in the presence of lipopolysaccharide is also increased in Stat6 macrophages and to a lesser extent, in Pparδ/γ macrophages. In concert, β-adrenergic agonist-induced lipolysis results in higher levels of cell death and inflammatory markers in ATMs derived from myeloid-specific Pparδ/γ or Stat6 mice.Our data suggest that ATM cell death is closely linked to metabolic inflammation. Within WAT where concentrations of free fatty acids fluctuate, M2 polarization regulated by the Stat6-Ppar axis enhances ATM\'s tolerance to lipid-mediated stress, thereby maintaining the homeostatic state.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: obesity

Priming of Hypothalamic Ghrelin Signaling and Microglia Activation Exacerbate Feeding in Rats\' Offspring Following Maternal Overnutrition.

Maternal overnutrition during pregnancy leads to metabolic alterations, including , hyperphagia, and inflammation in the offspring. Nutritional priming of central inflammation and its role in ghrelin sensitivity during fed and fasted states have not been analyzed. The current study aims to identify the effect of maternal programming on microglia activation and ghrelin-induced activation of hypothalamic neurons leading to food intake response. We employed a nutritional programming model exposing female Wistar rats to a cafeteria diet (CAF) from pre-pregnancy to weaning. Food intake in male offspring was determined daily after fasting and subcutaneous injection of ghrelin. Hypothalamic ghrelin sensitivity and microglia activation was evaluated using immunodetection for Iba-1 and c-Fos markers, and Western blot for TBK1 signaling. Release of TNF-alpha, IL-6, and IL-1β after stimulation with , oleic, linoleic , or C6 ceramide in primary microglia culture were quantified using ELISA. We found that programmed offspring by CAF diet exhibits overfeeding after fasting and peripheral ghrelin administration, which correlates with an increase in the hypothalamic Iba-1 microglia marker and c-Fos cell activation. Additionally, in contrast to oleic, linoleic, or C6 ceramide stimulation in primary microglia culture, stimulation with for 24 h promotes TNF-alpha, IL-6, and IL-1β release and TBK1 activation. Notably, intracerebroventricular (i.c.v.) or LPS inoculation for five days promotes daily increase in food intake and food consumption after ghrelin administration. Finally, we found that i.c.v. substantially activates hypothalamic Iba-1 microglia marker and c-Fos. Together, our results suggest that maternal nutritional programing primes ghrelin sensitivity and microglia activation, which potentially might mirror hypothalamic administration of the saturated .

Keyword: obesity

Integrating Extracellular Flux Measurements and Genome-Scale Modeling Reveals Differences between Brown and White Adipocytes.

White adipocytes are specialized for energy storage, whereas brown adipocytes are specialized for energy expenditure. Explicating this difference can help identify therapeutic targets for . A common tool to assess metabolic differences between such cells is the Seahorse Extracellular Flux (XF) Analyzer, which measures oxygen consumption and media acidification in the presence of different substrates and perturbagens. Here, we integrate the Analyzer\'s metabolic profile from human white and brown adipocytes with a genome-scale metabolic model to predict flux differences across the metabolic map. Predictions matched experimental data for the metabolite 4-aminobutyrate, the protein ABAT, and the fluxes for glucose, glutamine, and palmitate. We also uncovered a difference in how adipocytes dispose of nitrogenous waste, with brown adipocytes secreting less ammonia and more urea than white adipocytes. Thus, the method and software we developed allow for broader metabolic phenotyping and provide a distinct approach to uncovering metabolic differences.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: obesity

Hyperlipidemia-induced hepassocin in the liver contributes to insulin resistance in skeletal muscle.

Hepassocin (HPS) has recently been identified as a novel hepatokine that causes hepatic steatosis. However, the role of HPS in the development of insulin resistance in skeletal muscle under remains unclear. The effect of hyperlipidemia on hepatic HPS expression was evaluated in primary hepatocytes and liver of mice. HPS-mediated signal pathways were explored using small interfering (si) RNAs of specific genes or inhibitors. We found that treatment of primary hepatocytes with palmitate could induce HPS expression through C/EBPβ-mediated transcriptional activation. Furthermore, increased HPS expression was observed in the liver of high fat diet (HFD)-fed or tunicamycin-treated mice. Pretreatment with 4-phenylbutyrate (4-BPA) (an endoplasmic reticulum (ER) stress inhibitor) and suppression of p38 by siRNA abrogated the effect of palmitate on HPS expression in primary hepatocytes. Treatment of differentiated C2C12\xa0cells with recombinant HPS caused c-Jun N-terminal kinase (JNK) phosphorylation and impairment of insulin sensitivity in a dose-dependent manner. siRNA-mediated suppression of JNK reduced the effect of HPS on insulin signaling. Furthermore, the suppression of epidermal growth factor receptor (EGFR) by siRNA mitigated both HPS-induced JNK phosphorylation and insulin resistance. In addition, HPS did not affect inflammation and ER stress in differentiated C2C12\xa0cells. In conclusion, we elucidated that ER stress induced by palmitate could increase the expression of HPS in hepatocytes and further contribute to the development of insulin resistance in skeletal muscle via EGFR/JNK-mediated pathway. Taken together, we suggest that HPS could be a therapeutic target for -linked insulin resistance.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: obesity

Saturated fatty palmitate negatively regulates autophagy by promoting ATG5 protein degradation in meniscus cells.

and associated metabolic factors are major risk factors for the development of osteoarthritis. Previously, we have shown that the free fatty palmitate induces endoplasmic reticulum (ER) stress and induces apoptosis in meniscus cells. However, the molecular mechanisms involved in these effects are not clearly understood. In our current study, we found that palmitate inhibits autophagy by modulating the protein levels of autophagy-related genes-5 (ATG5) that is associated with decreased lipidation of LC3 and increased activation of cleaved caspase 3. Pretreatment of meniscus cells with 4-phenyl butyric , a small molecule chemical chaperone that alleviates ER stress, or with MG-132, a proteasome inhibitor, restored normal levels of ATG5 and autophagosome formation, and decreased expression of cleaved caspase 3. Thus, our data suggest that palmitate downregulates autophagy in meniscus cells by degrading ATG5 protein via ER-associated protein degradation, and thus promotes apoptosis. This is the first study to demonstrate that palmitate-induced endoplasmic reticulum stress negatively regulates autophagy.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: obesity

Sulforaphane ameliorates high-fat diet-induced spermatogenic deficiency in mice†.

Sulforaphane (SFN), a dietary isothiocyanate that is mainly found in cruciferous vegetables, possesses anti-oxidative and anticancer activity and modulates inflammation. However, little is known about the role of SFN in -related male reproductive defects. The present study aimed to investigate the effects of SFN on high-fat diet (HFD)-induced male spermatogenic impairment and further clarify the possible underlying mechanisms. In this study, 8-week-old mice were randomly divided into four groups. Mice were fed a normal diet or an HFD with or without SFN supplementation. Sulforaphane was subcutaneously injected at a dose of 0.5 mg/kg 5 days/week for 4 weeks beginning 8 weeks after initiation of the HFD. The results demonstrated that SFN could protect against HFD-induced reproductive dysfunction in male mice. Moreover, SFN also improved reproductive ability, as demonstrated by an increased pregnancy rate and decreased embryo resorption rate in comparison to the corresponding HFD group. We also observed a decrease in apoptosis and an attenuation of endoplasmic reticulum (ER) stress after SFN treatment. In vitro studies of mouse and human sperm samples also revealed that SFN protects against the -induced reduction in sperm viability and motility by inhibiting ER stress in an AMP-activated protein kinase (AMPK)-dependent manner. AMPK-dependent ER stress attenuation by SFN was further confirmed using AMPK knockout mice. Taken together, these data show that SFN protects against HFD-induced male reproductive dysfunction by inhibiting ER stress and apoptosis. These findings may be helpful for identifying new therapeutic methods to treat male infertility.© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

Keyword: obesity

Saturated fatty acids, , and the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in asthmatic patients.

Both and high dietary fat intake activate the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome.We aimed to examine NLRP3 inflammasome activity in the airways of obese asthmatic patients after macronutrient overload and in immune cells challenged by inflammasome triggers.Study 1 was a cross-sectional observational study of nonobese (n\xa0=\xa051) and obese (n\xa0=\xa076) asthmatic adults. Study 2 was a randomized, crossover, acute feeding study in 23 asthmatic adults (n\xa0=\xa012 nonobese and n\xa0=\xa011 obese subjects). Subjects consumed 3 isocaloric meals on 3 separate occasions (ie, saturated fatty , n-6 polyunsaturated fatty , and carbohydrate) and were assessed at 0 and 4\xa0hours. For Studies 1 and 2, airway inflammation was measured based on sputum differential cell counts, IL-1β protein levels (ELISA), and sputum cell gene expression (Nanostring nCounter). In Study 3 peripheral blood neutrophils and monocytes were isolated by using Ficoll density gradient and magnetic bead separation and incubated with or without , LPS, or TNF-α for 24\xa0hours, and IL-1β release was measured (ELISA).In Study 1 NLRP3 and nucleotide oligomerization domain 1 (NOD1) gene expression was upregulated, and sputum IL-1β protein levels were greater in obese versus nonobese asthmatic patients. In Study 2 the saturated fatty meal led to increases in sputum neutrophil percentages and sputum cell gene expression of Toll-like receptor 4 (TLR4) and NLRP3 at 4\xa0hours in nonobese asthmatic patients. In Study 3 neutrophils and monocytes released IL-1β when challenged with a combination of and LPS or TNF-α.The NLRP3 inflammasome is a potential therapeutic target in asthmatic patients. Behavioral interventions that reduce fatty exposure, such as weight loss and dietary saturated fat restriction, warrant further exploration.Copyright © 2018 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Keyword: obesity

Increased serum concentration of ceramides in obese children with nonalcoholic fatty liver disease.

Hepatic lipid accumulation is closely related to the development of insulin resistance, which is regarded as one of the most significant risk factors of nonalcoholic fatty liver disease (NAFLD). Although the exact molecular pathway leading to impaired insulin signaling has not been definitively established, ceramides are suspected mediators of lipid induced hepatic insulin resistance. Therefore, the aim of the study was to evaluate the serum ceramides concentration in obese children with NAFLD.The prospective study included 80 obese children (aged 7-17\xa0years, median 12\xa0years) admitted to our Department to diagnose initially suspected liver disease. Patients with viral hepatitis (HCV, HBV, CMV), autoimmune (AIH), toxic and metabolic (Wilson\'s disease, alfa-1-antitrypsin deficiency) liver diseases and celiac disease were excluded. NAFLD was diagnosed based on pediatric diagnostic criteria in obese children with liver steatosis in ultrasound (US) as well as elevated alanine transaminase (ALT) serum activity after exclusion of other major liver diseases listed before. Ultrasonography was used as a screening method and for qualitative assessment of the steatosis degree (graded according to Saverymuttu scale). Advanced steatosis was defined as a score\u2009>\u20091. The total intrahepatic lipid content (TILC) was assessed by magnetic resonance proton spectroscopy (HMRS) which is the most accurate technique for assessment of ectopic fat accumulation. Fasting serum concentration of ceramides was measured in 62 children.NAFLD was diagnosed in 31 children. Significant, positive correlation was found between total serum concentration of ceramides and insulin (r\xa0=\u20090.3, p\xa0=\u20090.02) and HOMA-IR (r\xa0=\u20090.28, p\xa0=\u20090.03). Total ceramide concentration as well as specific fatty -ceramides (FA-ceramides) concentrations, namely: myristic, , palmitoleic, stearic, oleic, behenic and lignoceric were significantly higher (p\xa0=\u20090.004, p\xa0=\u20090.003, p\xa0=\u20090.007, p\xa0<\u20090.001, p\xa0=\u20090.035, p\xa0=\u20090.008, p\xa0=\u20090.003, p\xa0=\u20090.006, respectively) in children with NAFLD compared to controls (n\xa0=\u200914). Moreover, children with NAFLD had significantly higher activity of ALT (p\xa0<\u20090.001) and GGT (p\xa0<\u20090.001), HOMA-IR (p\xa0=\u20090.04), BMI (p\xa0=\u20090.046), waist circumference (p\xa0=\u20090.01) steatosis grade in ultrasound (p\xa0<\u20090.001) and TILC in HMRS (p\xa0<\u20090.001) compared to children without NAFLD. We did not find significant differences in total and FA-ceramide species concentrations between children with mild (grade 1) and advanced liver steatosis in ultrasonography (grade 2-3).Elevated ceramide concentrations in obese patients together with their significant correlation with insulin resistance parameters suggest their association with molecular pathways involved in insulin signaling impairment known to be strongly linked to pathogenesis of non-alcoholic fatty liver disease.

Keyword: obesity

Protein translation associated to PERK arm is a new target for regulation of metainflammation: A connection with hepatocyte cholesterol.

Endoplasmic reticulum stress is a cellular phenomenon that has been associated with metabolic disorders, contributing to the development of , fatty liver disease, and dyslipidemias. Under metabolic overload conditions, in cells with a high protein-secretory activity, such as hepatocytes and Langerhans β cells, the unfolded protein response (UPR) is critical in to maintain protein homeostasis (proteostasis). UPR integrated by a tripartite signaling system, through activating transcription factor 6, protein kinase R-like endoplasmic reticulum kinase (PERK), and inositol-requiring enzyme 1, regulates gene transcription and translation to resolve stress and conserve proteostasis. In the current study, we demonstrated in hepatocytes under metabolic overload by saturated and stearic fatty acids, through activation of PERK signaling and CCAAT-enhancer-binding protein homologous protein (CHOP) transcription factor, an association with the expression of cyclooxygenase 2. More important, isolated exosomes from supernatants of macrophages exposed to lipopolysaccharides can also induce a metainflammation phenomenon, and when\xa0treated on hepatocytes, induced a rearrangement in cholesterol metabolism through sterol regulatory element-binding protein 2 (SREBP2), low-density lipoprotein receptor (LDLR), apolipoprotein A-I, and ABCA1. Moreover, we demonstrate the cellular effect of terpene-derived molecules, such as cryptotanshinone, isolated of plant Salvia brandegeei, regulating metainflammatory conditions through PERK pathway in both hepatocytes and β cells. Our data suggest the presence of a modulatory mechanism on specific protein translation process. This effect could be mediated by eukaryotic initiation factor-4A, evaluating salubrinal as a control molecule. Likewise, the protective mechanisms of unsaturated fatty acids, such as oleic and palmitoleic were confirmed. Therefore, modulation of metainflammation suggests a new target through PERK signaling in cells with a high secretory activity, and possibly the regulation of cholesterol in hepatocytes is promoted via exosomes.© 2018 Wiley Periodicals, Inc.

Keyword: obesity

Chronic intake of moderate fat-enriched diet induces fatty liver and low-grade inflammation without in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/ and metabolic syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: obesity

Vasoreactivity of isolated aortic rings from dyslipidemic and insulin resistant inducible nitric oxide synthase knockout mice.

Recent study from this lab indicated enhanced susceptibility of iNOS KO mice for diet induced (DIO) and systemic insulin resistance (IR) as compared to C57BL/6 (WT) mice. The present study investigates aortic vasoreactivity in high fat diet (HFD) induced insulin resistant iNOS KO mice. WT and iNOS KO mice were fed with 45% HFD/10% LFD for ten weeks. Systemic IR was assessed via measurement of circulating lipids, glucose, and insulin; while phenylephrine (PE)/acetylcholine (ACh) induced responses were monitored in the isolated aortic rings. To understand the mechanism, qPCR or Western blotting experiments were performed in aorta and Ea.hy926\u202fcells. After 10 weeks of HFD feeding, significant increase in the body weight/fat mass, augmented circulating lipids, glucose, insulin and inflammatory cytokines along with impaired acetylcholine induced aortic vasorelaxation and enhanced iNOS expression was observed in the aortic tissue of WT mice. In the aminoguanidine (AG, 20\u202fmg/kg for 4 weeks) treated WT mice and also in iNOS KO mice, acetylcholine induced vasorelaxation was significantly preserved. Further, acetylcholine mediated vasorelaxation correlated with increased eNOS phosphorylation at Ser1177 residue in the iNOS KO mice and same was also observed in the iNOS silenced Ea.hy926\u202fcells. Moreover, treatment of Ea.hy926\u202fcells with or TNFα also caused a significant decrease in eNOS activity, which was reversed in iNOS silenced Ea.hy926\u202fcells suggesting the role of iNOS in the reduction of eNOS activity. The study thus implies a critical role of iNOS in vascular diseases associated with dyslipidemia/IR.Copyright © 2019. Published by Elsevier B.V.

Keyword: obesity

Green Tea Polyphenol (-)-Epigallocatechin Gallate (EGCG) Attenuates Neuroinflammation in -Stimulated BV-2 Microglia and High-Fat Diet-Induced Obese Mice.

is closely associated with neuroinflammation in the hypothalamus, which is characterized by over-activated microglia and excessive production of pro-inflammatory cytokines. The present study was aimed at elucidating the effects of (-)-epigallocatechin gallate (EGCG) on -stimulated BV-2 microglia and high-fat-diet-induced obese mice. The results indicated the suppressive effect of EGCG on lipid accumulation, pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) release, and microglial activation in both cellular and high-fat-diet rodent models. These results were associated with lower phosphorylated levels of the janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) signaling pathway. In conclusion, EGCG can attenuate high-fat-induced hypothalamic inflammation via inhibiting the JAK2/STAT3 signaling pathways in microglia.

Keyword: obesity

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: obesity

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to -related insulin resistance and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including , insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: obesity

induces ceramide accumulation, mitochondrial protein hyperacetylation, and mitochondrial dysfunction in porcine oocytes.

Low oocyte quality is a possible causal factor of -induced infertility. High (PA) concentration in follicular fluid is a crucial feature noted in obese women. This study examined how high PA concentration reduced mitochondrial quality in oocytes and investigated a possible countermeasure against mitochondrial dysfunction. Cumulus cell-oocyte complexes were obtained from the ovaries of gilts, and incubated in medium containing PA (0.5 mM) or vehicle (BSA) for 44 h. Culturing oocytes at high PA concentration induced mitochondrial dysfunction determined by high reactive oxygen species and low ATP content in oocytes. Furthermore, high PA levels increased mitochondrial acetylation levels determined by a high degree of co-localization of TOMM20 and acetylated-lysine. In addition, high PA levels reduced the expression of Sirtuin 3 (SIRT3) and phosphorylated AMP-activated protein kinase (AMPK), while the AMPK activator, AICAR, restored mitochondrial function as well as oocyte ability and reduced the acetylation of mitochondrial protein. Supplementation of culture medium with dorsomorphin dihydrochloride (an AMPK inhibitor) reduced mitochondrial function and increased mitochondrial protein acetylation. Treatment of oocytes with LB100 (an inhibitor of AMPK dephosphorylation) reduced mitochondrial acetylation levels and restored mitochondrial function. Furthermore, high PA levels increased ceramide accumulation in oocytes, and addition of ceramide to the culture medium also induced mitochondrial dysfunction and increased mitochondrial acetylation. This detrimental effect of ceramide was diminished by AICAR treatment of oocytes. Our results indicated that PA induces ceramide accumulation and downregulates the AMPK/SIRT3 pathway causing mitochondrial protein hyperacetylation and dysfunction in oocytes.

Keyword: obesity

Loss of biliverdin reductase-A promotes lipid accumulation and lipotoxicity in mouse proximal tubule cells.

and increased lipid availability have been implicated in the development and progression of chronic kidney disease. One of the major sites of renal lipid accumulation is in the proximal tubule cells of the kidney, suggesting that these cells may be susceptible to lipotoxicity. We previously demonstrated that loss of hepatic biliverdin reductase A (BVRA) causes fat accumulation in livers of mice on a high-fat diet. To determine the role of BVRA in mouse proximal tubule cells, we generated a CRISPR targeting BVRA for a knockout in mouse proximal tubule cells (BVRA KO). The BVRA KO cells had significantly less metabolic potential and mitochondrial respiration, which was exacerbated by treatment with , a saturated fatty . The BVRA KO cells also showed increased intracellular triglycerides which were associated with higher fatty uptake gene cluster of differentiation 36 as well as increased de novo lipogenesis as measured by higher neutral lipids. Additionally, neutrophil gelatinase-associated lipocalin 1 expression, annexin-V FITC staining, and lactate dehydrogenase assays all demonstrated that BVRA KO cells are more sensitive to -induced lipotoxicity than wild-type cells. Phosphorylation of BAD which plays a role in cell survival pathways, was significantly reduced in -treated BVRA KO cells. These data demonstrate the protective role of BVRA in proximal tubule cells against saturated fatty -induced lipotoxicity and suggest that activating BVRA could provide a benefit in protecting from -induced kidney injury.

Keyword: obesity

Blockade of myeloid differentiation protein 2 prevents -induced inflammation and nephropathy.

is a major and independent risk factor of kidney diseases. The pathogenic mechanisms of -associated renal injury are recognized to at least involve a lipid-rich and pro-inflammatory state of the renal tissues, but specific mechanisms establishing causal relation remain unknown. Saturated fatty acids are elevated in , and known to induce chronic inflammation in kidneys. Myeloid differentiation protein 2 (MD2) is an important protein in lipopolysaccharide-induced innate immunity response and inflammation. We suggested that -associated renal injury is regulated by MD2 thereby driving an inflammatory renal injury. The used three mouse models for in vivo study: MD2 knockout mice (KO) maintained on high fat diet (HFD), wild-type mice on HFD plus L6H21, a specific MD2 inhibitor and KO mice given (PA) by IV injection. The in vitro studies were carried out in cultured renal tubular epithelial cells, mouse mesangial cells and primary macrophages, respectively. The HFD mice presented with increased hyperlipidemia, serum creatinine and proteinuria. Renal tissue from HFD mice had increased fibrosis, inflammatory cytokines, macrophage infiltration, and activation of NF-κB and MAPKs. This HFD-induced renal injury profile was not observed in KO mice or L6H21-treated mice. Mice given PA mimmicked the HFD-induced renal injury profiles, which were prevented by MD2 knockout. The in vitro data further confirmed MD2 mediates PA-induced inflammation. MD2 is causally related with -associated renal inflammatory injury. We believe that MD2 is an attractive target for future therapeutic strategies in -associated kidney diseases.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: obesity

A high-fat diet promotes depression-like behavior in mice by suppressing hypothalamic PKA signaling.

is associated with an increased risk of depression. The aim of the present study was to investigate whether is a causative factor for the development of depression and what is the molecular pathway(s) that link these two disorders. Using lipidomic and transcriptomic methods, we identified a mechanism that links exposure to a high-fat diet (HFD) in mice with alterations in hypothalamic function that lead to depression. Consumption of an HFD selectively induced accumulation of in the hypothalamus, suppressed the 3\', 5\'-cyclic AMP (cAMP)/protein kinase A (PKA) signaling pathway, and increased the concentration of free fatty receptor 1 (FFAR1). Deficiency of phosphodiesterase 4A (PDE4A), an enzyme that degrades cAMP and modulates stimulatory regulative G protein (Gs)-coupled G protein-coupled receptor signaling, protected animals either from genetic- or dietary-induced depression phenotype. These findings suggest that dietary intake of saturated fats disrupts hypothalamic functions by suppressing cAMP/PKA signaling through activation of PDE4A. FFAR1 inhibition and/or an increase of cAMP signaling in the hypothalamus could offer potential therapeutic targets to counteract the effects of dietary or genetically induced on depression.

Keyword: obesity

MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue inflammation in . Both MIP-1α and free fatty acids are elevated in /T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling pathways were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling pathways. In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in /T2D may contribute to metabolic inflammation through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: obesity

Signal transducer and activator of transcription-3 drives the high-fat diet-associated prostate cancer growth.

Prostate cancer (PCa) is the second leading cause of cancer death in men. PCa progression can be associated with . Signal transducer and activator of transcription-3 (STAT3) plays a crucial role in PCa growth. However, whether STAT3 plays a role in high-fat diet (HFD)-associated PCa growth is unknown. Our data show that HFD feeding increases tumor size, STAT3 phosphorylation, and (PA) level in the xenograft tissues of the PCa-bearing xenograft mouse model. In vitro studies show that PA increases STAT3 expression and phosphorylation (STAT3-Y705) in PCa. Computational modeling suggests strong and stable binding between PA and unphosphorylated STAT3 at R593 and N538. The binding changes STAT3 structure and activity. Functional studies show that both STAT3 mutants (R583A and N538A) and STAT3 dominant negative significantly reduce PA-enhanced STAT3 phosphorylation, PA-increased PCa cell proliferation, migration, and invasion. In the xenograft mouse models, the HFD-increased tumor growth and STAT3 phosphorylation in tumors are reversed by STAT3 inhibition. Our study not only demonstrates the regulatory role of PA/STAT3 axis in HFD-associated PCa growth but also suggests a novel mechanism of how STAT3 is activated by PA. Our data suggest STAT3 as a therapeutic target for the treatment of HFD-associated PCa.

Keyword: obesity

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of , insulin resistance, type-2 diabetes and metabolic syndrome. Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal metabolic disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA metabolism on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: obesity

Sphingosine-1-phosphate analog FTY720 reverses but not age-induced anabolic resistance to muscle contraction.

Sarcopenia, the age-associated loss of skeletal muscle mass and function, is coupled with declines in physical functioning leading to subsequent higher rates of disability, frailty, morbidity, and mortality. Aging and independently contribute to muscle atrophy that is assumed to be a result of the activation of mutual physiological pathways. Understanding mechanisms contributing to the induction of skeletal muscle atrophy with aging and is important for determining targets that may have pivotal roles in muscle loss in these conditions. We find that aging and equally induce an anabolic resistance to acute skeletal muscle contraction as observed with decreases in anabolic signaling activation after contraction. Furthermore, treatment with the sphingosine-1-phosphate analog FTY720 for 4 wk increased lean mass and strength, and the anabolic signaling response to contraction was improved in obese but not older animals. To determine the role of chronic inflammation and different fatty acids on anabolic resistance in skeletal muscle cells, we overexpressed IKKβ with and without exposure to saturated fatty (SFA; ), polyunsaturated fatty (eicosapentaenoic ), and monounsaturated fatty (oleic ). We found that IKKβ overexpression increased inflammation markers in muscle cells, and this chronic inflammation exacerbated anabolic resistance in response to SFA. Pretreatment with FTY720 reversed the inflammatory effects of in the muscle cells. Taken together, these data demonstrate chronic inflammation can induce anabolic resistance, SFA aggravates these effects, and FTY720 can reverse this by decreasing ceramide accumulation in skeletal muscle.

Keyword: obesity

Inhibition of cholinergic potentiation of insulin secretion from pancreatic islets by chronic elevation of glucose and fatty acids: Protection by casein kinase 2 inhibitor.

Chronic hyperlipidemia and hyperglycemia are characteristic features of type 2 diabetes (T2DM) that are thought to cause or contribute to β-cell dysfunction by "glucolipotoxicity." Previously we have shown that acute treatment of pancreatic islets with fatty acids (FA) decreases acetylcholine-potentiated insulin secretion. This acetylcholine response is mediated by M3 muscarinic receptors, which play a key role in regulating β-cell function. Here we examine whether chronic FA exposure also inhibits acetylcholine-potentiated insulin secretion using mouse and human islets.Islets were cultured for 3 or 4 days at different glucose concentration with 0.5\xa0mM (PA) or a 2:1 mixture of PA and oleic (OA) at 1% albumin (PA/BSA molar ratio 3.3). Afterwards, the response to glucose and acetylcholine were studied in perifusion experiments.FA-induced impairment of insulin secretion and Ca signaling depended strongly on the glucose concentrations of the culture medium. PA and OA in combination reduced acetylcholine potentiation of insulin secretion more than PA alone, both in mouse and human islets, with no evidence of a protective role of OA. In contrast, lipotoxicity was not observed with islets cultured for 3 days in medium containing less than 1\xa0mM glucose and a mixture of glutamine and leucine (7\xa0mM each). High glucose and FAs reduced endoplasmic reticulum (ER) Ca storage capacity; however, preserving ER Ca by blocking the IP3 receptor with xestospongin C did not protect islets from glucolipotoxic effects on insulin secretion. In contrast, an inhibitor of casein kinase 2 (CK2) protected the glucose dependent acetylcholine potentiation of insulin secretion in mouse and human islets against glucolipotoxicity.These results show that chronic FA treatment decreases acetylcholine potentiation of insulin secretion and that this effect is strictly glucose dependent and might involve CK2 phosphorylation of β-cell M3 muscarinic receptors.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: obesity

ACSL5 genotype influence on fatty metabolism: a cellular, tissue, and whole-body study.

Acyl-CoA Synthetase Long Chain 5 (ACSL5) gene\'s rs2419621 T/C polymorphism was associated with ACSL5 mRNA expression and response to lifestyle interventions. However, the mechanistic understanding of the increased response in T allele carriers is lacking. Study objectives were to investigate the effect of rs2419621 genotype and ACSL5 human protein isoforms on fatty oxidation and respiration.Human ACSL5 overexpression in C2C12 mouse myoblasts was conducted to measure C oxidation and protein isoform localization in vitro. C oxidation studies and Western blot analysis of ACSL5 proteins were carried out in rectus abdominis primary myotubes from 5 rs2419621 T allele carriers and 4 non-carriers. In addition, mitochondrial high-resolution respirometry was conducted on vastus lateralis muscle biopsies from 4 rs2419621 T allele carriers and 4 non-carriers. Multiple linear regression analysis was conducted to test the association between rs2419621 genotype and respiratory quotient related pre- and post-lifestyle intervention measurements in postmenopausal women with overweight or .In comparison to rs2419621 non-carriers, T allele carriers displayed higher levels of i) 683aa ACSL5 isoform, localized mainly in the mitochondria, playing a greater role in fatty oxidation in comparison to the 739aa protein isoform ii) in vitro CO production in rectus abdominis primary myotubes iii) in vivo fatty oxidation and lower carbohydrate oxidation post-intervention iv) ex vivo complex I and II tissue respiration in vastus lateralis muscle.These results support the conclusion that rs2419621 T allele carriers, are more responsive to lifestyle interventions partly due to an increase in the short ACSL5 protein isoform, increasing cellular, tissue and whole-body fatty utilization. With the increasing effort to develop personalized medicine to combat , our findings provide additional insight into genotypes that can significantly affect whole body metabolism and response to lifestyle interventions.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High-Fat-Diet-Induced Cardiac Dysfunction and Insulin Resistance by Elevating Tafazzin in Mouse Hearts.

High fat (HF)-diet-induced insulin resistance is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac insulin signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low fat (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs insulin signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the mitochondria. Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores insulin sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced insulin resistance further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to insulin resistance via mediating Drp-1 translocation to the mitochondria, and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates insulin resistance.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: obesity

Sex differences in the phagocytic and migratory activity of microglia and their impairment by .

Sex differences in the incidence, clinical manifestation, disease course, and prognosis of neurological diseases, such as autism spectrum disorders or Alzheimer\'s disease, have been reported. has been postulated as a risk factor for cognitive decline and Alzheimer\'s disease and, during pregnancy, increases the risk of autism spectrum disorders in the offspring. is associated with increased serum and brain levels of free fatty acids, such as , which activate microglial cells triggering a potent inflammatory cascade. In this study, we have determined the effect of in the inflammatory profile, motility, and phagocytosis of primary male and female microglia, both in basal conditions and in the presence of a pro-inflammatory stimulus (interferon-γ). Male microglia in vitro showed higher migration than female microglia under basal and stimulated conditions. In contrast, female microglia had higher basal and stimulated phagocytic activity than male microglia. did not affect basal migration or phagocytosis, but abolished the migration and phagocytic activity of male and female microglia in response to interferon-γ. These findings extend previous observations of sex differences in microglia and suggest that impairs the protective responses of these cells.© 2017 Wiley Periodicals, Inc.

Keyword: obesity

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro diabetes research.

The interplay between hyper-glycemia and -lipidemia in diabetes mellitus (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for lipid transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: obesity

Adiponectin homolog novel osmotin protects /diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in -associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against /diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: obesity

Exacerbation and Prolongation of Psoriasiform Inflammation in Diabetic Obese Mice: A Synergistic Role of CXCL5 and Endoplasmic Reticulum Stress.

Accumulating evidence suggests that psoriasis is frequently accompanied by metabolic disorders, such as and diabetes. However, the mechanisms underlying the association between increased psoriasis severity and concomitant metabolic syndrome have not been fully clarified. Herein, we show that imiquimod-induced psoriasiform inflammation was exacerbated and prolonged in diabetic obese mice compared to that in control mice, accompanied by remarkably increased lesional expressions of Cxcl5 and Il-1b. Notably, a large number of CXCL5 Ly6G cells infiltrated the dermis and subcutaneous fat tissue of the diabetic obese mice. Most macrophages in the subcutaneous fat tissues of the diabetic obese mice were positive for expression of IL-1β and GRP78/Bip, an endoplasmic reticulum stress marker. Depletion of Ly6G cells and macrophages diminished the imiquimod-induced psoriasiform inflammation. Further, CXCL5 potentiated the secretion of IL-1β from macrophages and , a fatty released from subcutaneous adipocytes, further enhanced IL-1β secretion via endoplasmic reticulum stress induction. Combined with the fact that the serum levels of both CXCL5 and are significantly elevated in patients with metabolic syndrome, our results suggest a role for CXCL5 and endoplasmic reticulum stress in the increase of psoriasis severity of patients with concomitant metabolic syndrome.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: obesity

Oleic ameliorates -induced ER stress and inflammation markers in naive and cerulein-treated exocrine pancreas cells.

Dietary fat overload (typical to ) increases the risk of pancreatic pathologies through mechanisms yet to be defined. We previously showed that saturated dietary fat induces pancreatic acinar lipotoxicity and cellular stress. The endoplasmic reticulum (ER) of exocrine pancreas cells is highly developed and thus predisposed to stress. We studied the combination of saturated and unsaturated FAs in metabolic and pancreatitis like cerulein (CER)-induced stress states on cellular ER stress.Exocrine pancreas AR42J and rat primary exocrine acinar cells underwent acute (24 h) challenge with different FAs (saturated, monounsaturated) at different concentrations (250 and 500 µM) and in combination with acute CER-induced stress, and were analyzed for fat accumulation, ER stress unfolded protein response (UPR) and immune and enzyme markers. Acute exposure of AR42J and pancreatic acinar cells to different FAs and their combinations increased triglyceride accumulation. significantly dose-dependently enhanced the UPR, immune factors and pancreatic lipase (PL) levels, as demonstrated by XBP1 splicing and elevation in UPR transcripts and protein levels ( and ). Exposure to high levels in a CER-induced stress state synergistically increased ER stress and inflammation marker levels. Exposure to oleic did not induce ER stress and PL levels and significantly decreased immune factors in an acute CER-induced stress state. Combination of oleic and acids significantly reduced the -induced ER stress, but did not affect the immune factor response. We show that combination of monounsaturated and saturated FAs protects from exocrine pancreatic cellular ER stress in both metabolic and CER-induced stress.© 2019 The Author(s).

Keyword: obesity

Impact of miR-140 Deficiency on Non-Alcoholic Fatty Liver Disease.

We have previously shown that loss of miR-140 has a pro-fibrotic effect in the mammary gland. This study aims to investigate whether miR-140 loss and act synergistically to promote non-alcoholic fatty liver disease (NAFLD), and to identify the underlying mechanisms.Liver tissues were isolated from lean-fat-diet and high-fat-diet fed wild-type and miR-140 knockout mice. Using molecular staining and immunohistochemistry techniques, increased development of NAFLD and fibrotic indicators in miR-140 knockout mice were identified. Utilizing an in vitro model system, miR-140 was demonstrated to target TLR-4, and miR-140 overexpression was shown to be sufficient to inhibit signaling through the TLR-4/NFκB pathway.These findings demonstrate that loss of miR-140 results in increased expression of TLR-4, sensitizing cells to signaling and in increased inflammatory activity through the TLR4/NFκB pathway. This signaling axis promotes NAFLD development in a high-fat diet context and indicates the potential utility of miR-140 rescue as a therapeutic strategy in NAFLD.© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: obesity

Association between increased visceral fat area and alterations in plasma fatty profile in overweight subjects: a cross-sectional study.

Visceral fat accumulation in overweight status has been resulted in changes of fatty profiles. The fatty acids\xa0profiles can be altered by fatty desaturase; the activity\xa0of which is highly associated with and other metabolic diseases. We hypothesized that fatty composition, desaturase activity, and accumulation of visceral fat are interrelated. Thus, the aim of this study was to investigate the association between increased visceral fat area and alterations in plasma fatty profile in overweight subjects with different amounts of visceral fat.Healthy overweight subjects (25.0\xa0kg/m\xa0≤\u2009BMI\u2009<\u200930\xa0kg/m,\xa0n=232) were classified into lower (T1), middle (T2), and upper tertiles\xa0(T3) according to L4 visceral fat area (T1: <71.8\xa0cm, T2: 71.8\xa0cm-99.6\xa0cm, T3: >99.6\xa0cm).The T3 group showed higher amounts of cis-10-heptadecenoic and activity of C16 Δ9-desaturase and C18 Δ9-desaturase and lower activity of Δ5-desaturase than the T1 group. Additionally, the T3 group showed higher amounts of saturated fatty acids, myristic , , stearic , monounsaturated fatty acids, palmitoleic , oleic , n-6 polyunsaturated fatty acids, linoleic , dihomo-γ-linolenic , arachidonic , n-3 PUFAs, and docosapentaenoic than the T1 and T2 groups.This study indicates that greater than a certain area (>99.6\xa0cm) of visceral fat is needed to observe altered levels of individual fatty species and desaturase activities. The results suggest that increased activity of C16 Δ9-desaturase and C18 Δ9-desaturase in parallel with decreased Δ5-desaturase activity may be a causative factor in disturbed fatty metabolism.

Keyword: obesity

Increases Endothelin-1 Expression in Vascular Endothelial Cells through the Induction of Endoplasmic Reticulum Stress and Protein Kinase C Signaling.

We investigated the regulation of endothelin-1 (ET-1) expression in in vivo high-fat diet (HFD)-fed mice and in vitro cultured human aortic endothelial cells (HAECs).Male C57BL/6 mice were fed on standard chow, serum was prepared, and ET-1 levels were analyzed using an ELISA kit. Quantitative PCR was performed using iQ SYBR Green Supermix. Statistical significance was assessed using SPSS, with p < 0.05 considered significant.The serum ET-1 content and endothelial expression of ET-1 mRNA were increased in the HFD-fed mice compared to the chow-fed control mice. Moreover, the mRNA expression of ET-1 was significantly increased in cultured HAECs in response to acute (< 24 h) and chronic (12-16 days) treatments with (PA), one of the most abundant saturated fatty acids in . We found that the induction of ET-1 expression by PA was abolished by pretreating the cells with the endoplasmic reticulum (ER) stress inhibitor 4-phenylbutyric or the protein kinase C (PKC) inhibitor Gö 6850.Our findings demonstrate for the first time that PA increases ET-1 expression in endothelial cells through the induction of ER stress and the activation of PKC, providing novel mechanistic insights into the pathogenesis of -associated hypertension and cardiovascular diseases.© 2018 S. Karger AG, Basel.

Keyword: obesity

Fatty and Lipopolysaccharide Effect on Beta Cells Proteostasis and its Impact on Insulin Secretion.

Metabolic overload by saturated fatty acids (SFA), which comprises β-cell function, and impaired glucose-stimulated insulin secretion are frequently observed in patients suffering from and type 2 diabetes mellitus. The increase of intracellular Ca triggers insulin granule release, therefore several mechanisms regulate Ca efflux within the β-cells, among others, the plasma membrane Ca-ATPase (PMCA). In this work, we describe that lipotoxicity mediated mainly by the saturated (PA) (16C) is associated with loss of protein homeostasis (proteostasis) and potentially cell viability, a phenomenon that was induced to a lesser extent by stearic (18C), myristic (14C) and lauric (12C) acids. PA was localized on endoplasmic reticulum, activating arms of the unfolded protein response (UPR), as also promoted by lipopolysaccharides (LPS)-endotoxins. In particular, our findings demonstrate an alteration in PMCA1/4 expression caused by PA and LPS which trigger the UPR, affecting not only insulin release and contributing to β-cell mass reduction, but also increasing reactive nitrogen species. Nonetheless, stearic (SA) did not show these effects. Remarkably, the proteolytic degradation of PMCA1/4 prompted by PA and LPS was avoided by the action of monounsaturated fatty acids such as oleic and palmitoleic . Oleic recovered cell viability after treatment with PA/LPS and, more interestingly, relieved endoplasmic reticulum (ER) stress. While palmitoleic improved the insulin release, this fatty seems to have more relevant effects upon the expression of regulatory pumps of intracellular Ca. Therefore, chain length and unsaturation of fatty acids are determinant cues in proteostasis of β-cells and, consequently, on the regulation of calcium and insulin secretion.

Keyword: obesity

Myricanol modulates skeletal muscle-adipose tissue crosstalk to alleviate high-fat diet-induced and insulin resistance.

Skeletal muscle is the predominant site for glucose disposal and fatty consumption. Emerging evidence indicates that the crosstalk between adipose tissue and skeletal muscle is critical in maintaining insulin sensitivity and lipid homeostasis. The current study was designed to investigate whether myricanol improves insulin sensitivity and alleviates adiposity through modulating skeletal muscle-adipose tissue crosstalk.The therapeutic effect of myricanol was evaluated on (PA)-treated C2C12 myotubes and high-fat diet (HFD)-fed mice. The crosstalk between myotubes and adipocytes was evaluated using Transwell assay. The cellular lipid content was examined by Nile red staining. The mitochondrial content was assessed by MitoTracker Green staining and citrate synthase activity, and the mitochondrial function was examined by Seahorse assay. Expression of mitochondria-related and insulin signalling pathway proteins was analysed by Western blot, and the irisin level was determined by elisa kit.Myricanol increased mitochondrial quantity and function through activating AMP-activated protein kinase, resulting in reduced lipid accumulation and enhanced insulin-stimulated glucose uptake, in PA-treated C2C12 myotubes. Furthermore, myricanol stimulated irisin production and secretion from myotubes to reduce lipid content in 3T3-L1 adipocytes. In HFD-fed mice, myricanol treatment alleviated adiposity and insulin resistance through enhancing lipid utilization and irisin production in skeletal muscle and inducing browning of inguinal fat.Myricanol modulates skeletal muscle-adipose tissue crosstalk, to stimulate browning of adipose tissue and improve insulin sensitivity in skeletal muscle. Myricanol might be a potential candidate for treating insulin resistance and .© 2019 The British Pharmacological Society.

Keyword: obesity

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: obesity

Palmitate and oleate co-treatment increases myocellular protein content via impaired protein degradation.

Protein balance is a crucial determinant of myocellular size and function. The effects of fatty acids on myocellular protein balance remain controversial. The aim of this study was to determine the direct effects of a mixed-species fatty environment on myocellular protein synthesis and degradation.C2C12 myotubes were cultured in media containing equimolar (250\xa0μM) and oleate (PO) or bovine serum albumin control for ≤72\xa0h. Myocellular protein balance was determined via incorporation (synthesis) or release (degradation) of H-tyrosine after 24, 48, and 72\xa0h of treatment. Expression of major proteolytic genes was measured by reverse transcription polymerase chain reaction.PO significantly increased myocellular protein content at 24, 48, and 72\xa0h. Basal myocellular protein synthesis was unchanged by PO. However, PO significantly decreased basal rate of protein degradation at 24\xa0h and this effect persisted throughout 72\xa0h of treatment. Expression of the proteolytic genes Atrogin-1 (MAFbx), MuRF-1, LC3, and ATG4 B, was reduced during the 72\xa0h PO.A mixed-species fatty environment increases myocellular protein content by decreasing the rate of protein degradation, which may be regulated at the level of gene transcription.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: obesity

Adipokines secretion in feline primary adipose tissue culture in response to dietary fatty acids.

in cats has been associated with alterations in adipokines including: adiponectin, interleukin-6 (IL6), and tumor necrosis factor-α (TNFα). Omega-3 polyunsaturated fatty acids have multiple beneficial effects on -associated disorders, and therefore may alleviate these alterations. This study aimed to determine the effects of body condition, fat depot, troglitazone, and different fatty acids on secretion of adiponectin, IL6 and TNFα from adipose tissue of healthy cats. Subcutaneous and visceral adipose tissue samples were collected from 18 healthy intact female cats, and body condition score (Range 3-7/9) was determined. Concentrations of adiponectin were measured in mature adipocytes cultures and concentrations of IL6 and TNFα were measured in stromovascular cells cultures following treatment with control medium, troglitazone at 10\u2009μM, eicosapentaenoic , arachidonic , or , at 25, 50, or 100\u2009μM.Stromovascular cells of visceral origin secreted higher concentrations of IL6 than corresponding cells of subcutaneous origin (P\u2009=\u20090.003). Arachidonic treatment at 25, 50, and 100\u2009μM increased IL6 secretion in subcutaneous (P\u2009=\u20090.045, P\u2009=\u20090.002, and P\u2009<\u20090.001, respectively) and visceral (P\u2009=\u20090.034, P\u2009=\u20090.001, and P\u2009<\u20090.001, respectively) stromovascular cells. Eicosapentaenoic treatment increased TNFα secretion in subcutaneous stromovascular cells at 25, 50, and 100\u2009μM (P\u2009=\u20090.002, P\u2009=\u20090.001, and P\u2009=\u20090.015, respectively) and in visceral stromovascular cells at 50\u2009μM (P\u2009<\u20090.001). No significant effect on medium adiponectin concentration was observed following troglitazone treatment (P\u2009=\u20090.4) or fatty acids treatments at 25 (P\u2009=\u20090.2), 50 (P\u2009=\u20090.8), or 100 (P\u2009=\u20090.7) μM. Body condition score did not have significant effects on medium concentrations of adiponectin (P\u2009=\u20090.4), IL6 (P\u2009=\u20090.1), or TNFα (P\u2009=\u20090.8).This study demonstrated higher basal secretion of IL6 from visceral compared to subcutaneous adipose tissue, a stimulatory effect of arachidonic on secretion of IL6 and a stimulatory effect of eicosapentaenoic on TNFα from feline adipose tissue.

Keyword: obesity

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only energy metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating -induced inflammation.

Keyword: obesity

Dietary Fatty Acids Amplify Inflammatory Responses to Infection through p38 MAPK Signaling.

is an important risk factor for severe asthma exacerbations, which are mainly caused by respiratory infections. Dietary fatty acids, which are increased systemically in obese patients and are further increased after high-fat meals, affect the innate immune system and may contribute to dysfunctional immune responses to respiratory infection. In this study we investigated the effects of dietary fatty acids on immune responses to respiratory infection in pulmonary fibroblasts and a bronchial epithelial cell line (BEAS-2B). Cells were challenged with BSA-conjugated fatty acids (ω-6 polyunsaturated fatty acids [PUFAs], ω-3 PUFAs, or saturated fatty acids [SFAs]) +/- the viral mimic polyinosinic:polycytidylic (poly[I:C]) or bacterial compound lipoteichoic (LTA), and release of proinflammatory cytokines was measured. In both cell types, challenge with arachidonic (AA) (ω-6 PUFA) and poly(I:C) or LTA led to substantially greater IL-6 and CXCL8 release than either challenge alone, demonstrating synergy. In epithelial cells, (SFA) combined with poly(I:C) also led to greater IL-6 release. The underlying signaling pathways of AA and poly(I:C)- or LTA-induced cytokine release were examined using specific signaling inhibitors and IB. Cytokine production in pulmonary fibroblasts was prostaglandin dependent, and synergistic upregulation occurred via p38 mitogen-activated protein kinase signaling, whereas cytokine production in bronchial epithelial cell lines was mainly mediated through JNK and p38 mitogen-activated protein kinase signaling. We confirmed these findings using rhinovirus infection, demonstrating that AA enhances rhinovirus-induced cytokine release. This study suggests that during respiratory infection, increased levels of dietary ω-6 PUFAs and SFAs may lead to more severe airway inflammation and may contribute to and/or increase the severity of asthma exacerbations.

Keyword: obesity

Association Between Plasma N-Acylethanolamides and High Hemoglobin Concentration in Southern Peruvian Highlanders.

Alarcón-Yaquetto, Dulce E., Lidia Caballero, and Gustavo F. Gonzales. Association between plasma N-acylethanolamides and high hemoglobin concentration in Southern Peruvian highlanders. High Alt Med Biol 18:322-329, 2017.-High-altitude (HA) hypoxia is a stressful condition endured by organisms through different mechanisms. Failing to adapt to chronic HA exposure leads to a disease called chronic mountain sickness (CMS) characterized by excessive erythrocytosis (hemoglobin [Hb] ≥19\u2009g/dL for women and ≥21\u2009g/dL for men). Genes encoding for peroxisome proliferator-activated receptor (PPAR) subunits α and γ have been proposed as candidate genes for HA adaptation. N-acylethanolamides (NAEs) are endogenous fatty substances that bind to PPAR-α and -γ. NAEs are also able to modulate the endocannabinoid system, a signaling pathway activated in physiological stressful conditions. In the frame of a metabolomic study, we measured plasma levels of four NAEs: palmitoylethanolamide (PEA), oleoylethanolamide (OEA), stearoyl ethanolamide (SEA), and linoleoyl ethanolamide (LEA) in natives from Puno (3830\u2009m), a city located in the Peruvian Southern Andes, and Lima (150\u2009m). All NAEs were significantly higher in the HA population (p\u2009<\u20090.001, q\u2009<\u20090.001). Subjects with higher NAE values were those with higher Hb concentration and lower pulse saturation. However, there was no association between NAEs and CMS score. Our results suggest that PEA and OEA could be involved in physiological regulation following long-term HA exposure.

Keyword: oxygen

Cytochrome c release from rat liver mitochondria is compromised by increased saturated cardiolipin species induced by sucrose feeding.

Cytochrome c release from mitochondria has been described to be related to reactive species (ROS) generation. With ROS generation being increased in fatty liver from sucrose-fed (SF) rats, we hypothesized that cytochrome c release might be positively associated with H2O2 generation from SF mitochondria. Surprisingly, cytochrome c release from mitochondria of SF liver was found to be significantly lower compared with control (C) mitochondria oxidizing pyruvate/malate or succinate. Exposure of mitochondria to exogenous superoxide radical generated by the xanthine/xanthine oxidase system elicits a dose-response cytochrome c release in both control and SF mitochondria, but cytochrome c release remains lower in SF mitochondria compared with C mitochondria. Furthermore, the addition of ebselen, PEG-catalase, or catalase, a H2O2 scavenger, significantly reduces cytochrome c release from C and SF mitochondria. Our results suggest that both intra- and extramitochondrial H2O2 are involved in cytochrome c release, but the persisting difference between C and SF levels can be attributed to the differences in cardiolipin compositions. Indeed, the ratio of -rich cardiolipin species was found to be increased in lipid membrane from SF mitochondria compared with C mitochondria, whereas that of linoleic -rich cardiolipin species was found decreased. In addition, the content of tafazzin, a protein responsible for cardiolipin remodeling, was decreased in SF mitochondria. Therefore, we conclude that the changes observed in the composition of cardiolipin molecular species in SF mitochondria may be involved in cytochrome c interaction with mitochondrial inner membrane lipid and in its reduced release from SF mitochondria.Copyright © 2015 the American Physiological Society.

Keyword: oxygen

-Induced Podocyte Apoptosis via the Reactive Species-Dependent Mitochondrial Pathway.

Chronic kidney disease (CKD) is often accompanied by hyperlipidemia, which accelerates progression of the disease. Podocyte injury can lead to dysfunction of the glomerular filtration barrier, which is associated with proteinuria, a risk marker for the progression of CKD. Our previous studies demonstrated that (PA) can induce podocyte apoptosis; however, the underlying mechanisms are unclear. In the present study, we investigated the specific molecular mechanisms of PA-induced apoptosis in cultured podocytes.We cultured mouse podocytes and treated them with PA. Then, cell viability was measured using the Cell Counting Kit-8 colorimetric assay, lipid uptake was assessed by Oil Red O staining and boron-dipyrromethene staining, apoptosis was measured by flow cytometry, mitochondrial injury was assessed by JC-1 staining and transmission electron microscopy, and mitochondrial production of reactive species (ROS) was evaluated by fluorescence microscopy using the MitoSOX Red reagent. The effects of PA on the mitochondria-mediated caspase activation pathway were investigated by examining the expression of caspase-8, cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), B-cell lymphoma 2 (Bcl-2), Bax, Bid, cytochrome c, and Fas-associated protein with death domain (FADD) using western blotting. The translocation of Bax and cytochrome c were detected by immunofluorescence.PA treatment significantly increased lipid accumulation and induced podocyte apoptosis. We investigated whether the two primary apoptosis signaling pathways (death receptor-mediated pathway and mitochondria-mediated pathway) were involved in the execution of PA-induced podocyte apoptosis, and found that the levels of FADD, caspase-8, and Bid did not significantly change during this process. Meanwhile, PA treatment induced an increase in Bax protein expression and a decrease in Bcl-2 protein expression, with Bax translocation to the mitochondria. Furthermore, PA treatment induced mitochondrial impairment, and triggered the release of cytochrome c from the mitochondria to cytosol, with a concomitant dose-dependent increase in the levels of cleaved caspase-9, cleaved caspase-3, and PARP. Meanwhile, PA treatment increased mitochondrial production of ROS, and the mitochondria-targeted antioxidant mitoTEMPO significantly ameliorated PA-induced podocyte apoptosis.Our findings indicated that PA induced caspase-dependent podocyte apoptosis through the mitochondrial pathway, and mitochondrial ROS production participated in this process, thus potentially contributing to podocyte injury.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: oxygen

Dual roles of ULK1 (unc-51 like autophagy activating kinase 1) in cytoprotection against lipotoxicity.

Saturated fatty (SFA)-induced lipotoxicity is caused by the accumulation of reactive species (ROS), which is associated with damaged mitochondria. Moreover, lipotoxicity is crucial for the progression of nonalcoholic steatohepatitis (NASH). Autophagy is required for the clearance of protein aggregates or damaged mitochondria to maintain cellular metabolic homeostasis. The NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2)-KEAP1 (kelch like ECH associated protein 1) pathway is essential for the elimination of ROS. ULK1 (unc-51 like autophagy activating kinase 1; yeast Atg1) is involved in the initiation of autophagy; however, its role in lipotoxicity-induced cell death in hepatocytes and mouse liver has not been elucidated. We now show that ULK1 potentiates the interaction between KEAP1 and the autophagy adaptor protein SQSTM1/p62, thereby mediating NFE2L2 activation in a manner requiring SQSTM1-dependent autophagic KEAP1 degradation. Furthermore, ULK1 is required for the autophagic removal of damaged mitochondria and to enhance binding between SQSTM1 and PINK1 (PTEN induced kinase 1). This study demonstrates the molecular mechanisms underlying the cytoprotective role of ULK1 against lipotoxicity. Thus, ULK1 could represent a potential therapeutic target for the treatment of NASH. Abbreviations: ACTB: actin beta; CM-HDCFDA:5-(and-6)-chloromethyl-2\',7\'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; LPS: lipopolysaccharides; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAPK8/JNK: mitogen-activated protein kinase 8; MEF: mouse embryonic fibroblast; MFN1: mitofusin 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NASH: nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: ; PARP: poly (ADP-ribose) polymerase 1; PINK1: PTEN induced kinase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PRKC/PKC: protein kinase C; RBX1: ring-box 1; ROS: reactive species; SFA: saturated fatty ; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; TUBA: tubulin alpha; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase 1.

Keyword: oxygen

Trimer procyanidin oligomers contribute to the protective effects of cinnamon extracts on pancreatic β-cells in vitro.

Cinnamon extracts rich in procyanidin oligomers have shown to improve pancreatic β-cell function in diabetic db/db mice. The aim of this study was to identify the active compounds in extracts from two species of cinnamon responsible for the pancreatic β-cell protection in vitro.Cinnamon extracts were prepared from Cinnamomum tamala (CT-E) and Cinnamomum cassia (CC-E). Six compounds procyanidin B2 (cpd1), (-)-epicatechin (cpd2), cinnamtannin B1 (cpd3), procyanidin C1 (cpd4), parameritannin A1 (cpd5) and cinnamtannin D1 (cpd6) were isolated from the extracts. INS-1 pancreatic β-cells were exposed to (PA) or H2O2 to induce lipotoxicity and oxidative stress. Cell viability and apoptosis as well as ROS levels were assessed. Glucose-stimulated insulin secretion was examined in PA-treated β-cells and murine islets.CT-E, CC-E as well as the compounds, except cpd5, did not cause cytotoxicity in the β-cells up to the maximum dosage using in this experiment. CT-E and CC-E (12.5-50 μg/mL) dose-dependently increased cell viability in both PA- and H2O2-treated β-cells, and decreased ROS accumulation in H2O2-treated β-cells. CT-E caused more prominent β-cell protection than CC-E. Furthermore, CT-E (25 and 50 μg/mL) dose-dependently increased glucose-stimulated insulin secretion in PA-treated β-cells and murine islets, but CC-E had little effect. Among the 6 compounds, trimer procyanidins cpd3, cpd4 and cpd6 (12.5-50 μmol/L) dose-dependently increased the cell viability and decreased ROS accumulation in H2O2-treated β-cells. The trimer procyanidins also increased glucose-stimulated insulin secretion in PA-treated β-cells.Trimer procyanidins in the cinnamon extracts contribute to the pancreatic β-cell protection, thus to the anti-diabetic activity.

Keyword: oxygen

Cytotoxic effect of Kalanchoe flammea and induction of intrinsic mitochondrial apoptotic signaling in prostate cancer cells.

Kalanchoe flammea Stapf (Crassulaceae) is a medicinal plant grown in the South of Mexico (State of Tabasco), which is commonly used in traditional medicine for the treatment of fever, wounds, inflammation, and cancer.To establish the potential of K. flammea for the treatment of prostate cancer, evaluating its cytotoxic activity, its probable mechanism of action, and carrying out some toxicological safety studies.The cytotoxic activity of the ethyl acetate extract of K. flammea (Kf-EtOAc) was evaluated in several cell lines of prostate cancer by MTT viability assay. The cellular death mechanism was studied by evaluating the translocation of phosphatidylserine (Annexin V); overproduction of reactive species [2\'-7\'-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay]; release of Cytochrome C; activation of caspase-3 and\xa0-9, and regulation of Bcl-2, XIAP, and PKCε proteins by Western Blot analysis. For the evaluation of the safety of Kf-EtOAc, the Ames test, Micronucleus assay, and acute toxicity study were determined.Kf-EtOAc exhibited selective cytotoxic activity against prostate cell lines as follows: PC-3, LNCaP, and PrEC (IC = 1.36\u202f±\u202f0.05; 2.06\u202f±\u202f0.02, and 127.05\u202f±\u202f0.07\u202fμg/mL, respectively). The F82-P2 fraction (rich in coumaric and ) obtained by bioassay-guided fractionation of Kf-EtOAc also demonstrated selective cytotoxic activity against PC-3 cells (IC = 1.05\u202f±\u202f0.06\u202fμg/mL). Kf-EtOAc induces apoptosis by the intrinsic pathway; this mechanism of cell death was confirmed after observing that the extract produces phosphatidylserine translocation, overproduction of reactive species, release of Cytochrome C at mitochondrial level, and activation of caspase-3 and -9. It was also observed that Kf-EtOAc produces significant downregulation of apoptosis-related proteins Bcl-2, XIAP, and PKCε and induces DNA fragmentation and cell cycle arrest. In addition, Kf-EtOAc is non-genotoxic in vitro by Ames test and non-genotoxic in vivo by Micronucleus assay, and no signs of toxicity or death were reported after the administration of a single acute exposure of 2000\u202fmg/kg.K. flammea is a potential candidate for the development of new drugs for the treatment of prostate cancer. However, to propose their use in clinical trials, additional studies are required to understand their pharmacokinetic behavior, as well as the development of a suitable pharmaceutical form.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: oxygen

Beyond the Scavenging of Reactive Species (ROS): Direct Effect of Cerium Oxide Nanoparticles in Reducing Fatty Acids Content in an In Vitro Model of Hepatocellular Steatosis.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic accumulation of lipids. Antisteatotic effects of cerium oxide nanoparticles (CeONPs) have recently been shown in animal models of liver disease. However, it is unclear whether the activity of CeONPs is related solely to the decrease in oxidative stress or, in addition, they directly decrease liver fatty accumulation. To address this question, in this work, we used an in vitro model of hepatocellular steatosis, exposing HepG2 cells to oleic and . Cell uptake of CeONPs and their effect on oxidative stress and viability of hepatic cells cultured with HO were also evaluated. Results show that CeONPs were uptaken by HepG2 cells and reduced oxidative stress and improved cell viability. Treatment with oleic and increased lipogenesis and the content of different fatty acids. CeONPs reduced and stearic and most fatty acids consisting of more than 18 carbon atoms. These effects were associated with significant changes in elongase and desaturase activity. In conclusion, CeONPs directly protected HepG2 cells from cell injury in oxidative stress conditions and reduced fatty content in steatotic conditions by inducing specific changes in fatty metabolism, thus showing potential in the treatment of NAFLD.

Keyword: oxygen

The Mitochondria-Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

Mitochondria-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive species (ROS) production during glucose metabolism in β cells can be exacerbated under hyperglycaemic conditions such as type 2 diabetes (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: oxygen

Concerted action of p62 and Nrf2 protects cells from -induced lipotoxicity.

Nonalcoholic fatty liver disease (NAFLD), frequently associated with obesity and diabetes mellitus, is caused by the accumulation of excess fatty acids within liver cells. (PA), a common saturated fatty found in mammals, induces the generation of reactive species (ROS) and elicits apoptotic cell death, known as lipotoxicity. However, protective mechanisms against PA-induced lipotoxicity have not been elucidated. In this study, we aimed to clarify the role of p62, an adapter protein in the autophagic process, as well as the nuclear factor erythroid 2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) pathway, in protecting cells from PA-induced lipotoxicity. The Nrf2-Keap1 pathway is essential for the protection of cells from oxidative stress. p62 enhances its binding to Keap1 and leads to Nrf2 activation. Here, we show that PA potentiates Keap1 degradation and thereby activates the transcription of Nrf2 target genes partially through autophagy. Furthermore, this PA-mediated Keap1 degradation depends on p62. Correspondingly, a lack of p62 attenuates the PA-mediated Nrf2 activation and increases the susceptibility of cells to oxidative stress. These results indicate that p62 plays an important role in protecting cells against lipotoxicity through Keap1 degradation-mediated Nrf2 activation.Copyright © 2015 Elsevier Inc. All rights reserved.

Keyword: oxygen

Saturated lipids decrease mitofusin 2 leading to endoplasmic reticulum stress activation and insulin resistance in hypothalamic cells.

Endoplasmic reticulum (ER) and mitochondria dysfunction contribute to insulin resistance generation during obesity and diabetes. ER and mitochondria interact through Mitofusin 2 (MTF2), which anchors in the outer mitochondrial and ER membranes regulating energy metabolism. Ablation of MTF2 leads to ER stress activation and insulin resistance. Here we determine whether lipotoxic insult induced by saturated lipids decreases MTF2 expression leading to ER stress response in hypothalamus and its effects on insulin sensitivity using in vitro and in vivo models. We found that lipotoxic stimulation induced by , but not the monounsaturated palmitoleic , decreases MTF2 protein levels in hypothalamic mHypoA-CLU192 cells. Also, incubation activates ER stress response evidenced by increase in the protein levels of GRP78/BIP marker at later stage than MTF2 downregulation. Additionally, we found that MTF2 alterations induced by , but not palmitoleic, stimulation exacerbate insulin resistance in hypothalamic cells. Insulin resistance induced by is prevented by pre-incubation of the anti-inflammatory and the ER stress release reagents, sodium salicylate and 4 phenylbutirate, respectively. Finally, we demonstrated that lipotoxic insult induced by high fat feeding to mice decreases MTF2 proteins levels in arcuate nucleus of hypothalamus. Our data indicate that saturated lipids modulate MTF2 expression in hypothalamus coordinating the ER stress response and the susceptibility to insulin resistance.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: oxygen

Mesenchymal Stem Cells Ameliorated Glucolipotoxicity in HUVECs through TSG-6.

Glucolipotoxicity is one of the critical causal factors of diabetic complications. Whether mesenchymal stem cells (MSCs) have effects on glucolipotoxicity in human umbilical vein endothelial cells (HUVECs) and mechanisms involved are unclear. Thirty mM glucose plus 100 μM was used to induce glucolipotoxicity in HUVECs. MSCs and HUVECs were co-cultured at the ratio of 1:5 via Transwell system. The mRNA expressions of inflammatory factors were detected by RT-qPCR. The productions of reactive species (ROS), cell cycle and apoptosis were analyzed by flow cytometry. The tumor necrosis factor-α stimulated protein 6 (TSG-6) was knockdown in MSCs by RNA interference. High glucose and remarkably impaired cell viability and tube formation capacity, as well as increased the mRNA expression of inflammatory factors, ROS levels, and cell apoptosis in HUVECs. MSC co-cultivation ameliorated these detrimental effects in HUVECs, but no effect on ROS production. Moreover, TSG-6 was dramatically up-regulated by high glucose and fatty stimulation in both MSCs and HUVECs. TSG-6 knockdown partially abolished the protection mediated by MSCs. MSCs had protective effects on high glucose and induced glucolipotoxicity in HUVECs, and TSG-6 secreted by MSCs was likely to play an important role in this process.

Keyword: oxygen

CsrA regulates a metabolic switch from amino to glycerolipid metabolism.

CsrA plays a crucial role in the life-stage-specific expression of virulence phenotypes and metabolic activity. However, its exact role is only partly known. To elucidate how CsrA impacts metabolism we analysed the CsrA depended regulation of metabolic functions by comparative C-isotopologue profiling and consumption experiments of a wild-type (wt) strain and its isogenic mutant. We show that a mutant has significantly lower respiration rates when serine, alanine, pyruvate, α-ketoglutarate or palmitate is the sole carbon source. By contrast, when grown in glucose or glycerol, no differences in respiration were detected. Isotopologue profiling uncovered that the transfer of label from [U-C]serine via pyruvate into the citrate cycle and gluconeogenesis was lower in the mutant as judged from the labelling patterns of protein-derived amino acids, cell-wall-derived diaminopimelate, sugars and amino sugars and 3-hydroxybutyrate derived from polyhydroxybutyrate (PHB). Similarly, the incorporation of [U-C]glucose via the glycolysis/Entner-Doudoroff (ED) pathway but not via the pentose phosphate pathway was repressed in the mutant. On the other hand, fluxes due to [U-C]glycerol utilization were increased in the mutant. In addition, we showed that exogenous [1,2,3,4-C] is efficiently used for PHB synthesis via C-acetyl-CoA. Taken together, CsrA induces serine catabolism via the tricarboxylic cycle and glucose degradation via the ED pathway, but represses glycerol metabolism, fatty degradation and PHB biosynthesis, in particular during exponential growth. Thus, CsrA has a determining role in substrate usage and carbon partitioning during the life cycle and regulates a switch from amino usage in replicative phase to glycerolipid usage during transmissive growth.© 2017 The Authors.

Keyword: oxygen

Palmitate is not an effective fuel for pancreatic islets and amplifies insulin secretion independent of calcium release from endoplasmic reticulum.

The aim of the study was to determine the acute contribution of fuel oxidation in mediating the increase in insulin secretion rate (ISR) in response to fatty acids. Measures of mitochondrial metabolism, as reflected by consumption rate (OCR) and cytochrome c reduction, calcium signaling, and ISR by rat islets were used to evaluate processes stimulated by acute exposure to (PA). The contribution of mitochondrial oxidation of PA was determined in the presence and absence of a blocker of mitochondrial transport of fatty acids (etomoxir) at different glucose concentrations. Subsequent to increasing glucose from 3 to 20 mM, PA caused small increases in OCR and cytosolic calcium (about 20% of the effect of glucose). In contrast, the effect of PA on ISR was almost 3 times that by glucose, suggesting that the metabolism of PA is not the dominant mechanism mediating PA\'s effect on ISR. This was further supported by lack of inhibition of PA-stimulated OCR and ISR when blocking entry of PA into mitochondria (with etomoxir), and PA\'s lack of stimulation of reduced cytochrome c in the presence of high glucose. Consistent with the lack of metabolic stimulation by PA, an inhibitor of calcium release from the endoplasmic reticulum, but not a blocker of L-type calcium channels, abolished the PA-induced elevation of cytosolic calcium. Notably, ISR was unaffected by thapsigargin showing the dissociation of endoplasmic reticulum calcium release and second phase insulin secretion. In conclusion, stimulation of ISR by PA was mediated by mechanisms largely independent of the oxidation of the fuel.

Keyword: oxygen

The NOX1 isoform of NADPH oxidase is involved in dysfunction of liver sinusoids in nonalcoholic fatty liver disease.

The increased production of reactive species (ROS) has been postulated to play a key role in the progression of nonalcoholic fatty liver disease (NAFLD). However, the source of ROS and mechanisms underlying the development of NAFLD have yet to be established. We observed a significant up-regulation of a minor isoform of NADPH oxidase, NOX1, in the liver of nonalcoholic steatohepatitis (NASH) patients as well as of mice fed a high-fat and high-cholesterol (HFC) diet for 8 weeks. In mice deficient in Nox1 (Nox1KO), increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 demonstrated in HFC diet-fed wild-type mice (WT) were significantly attenuated. Concomitantly, increased protein nitrotyrosine adducts, a marker of peroxynitrite-induced injury detected in hepatic sinusoids of WT, were significantly suppressed in Nox1KO. The expression of NOX1 mRNA was much higher in the fractions of enriched liver sinusoidal endothelial cells (LSECs) than in those of hepatocytes. In primary cultured LSECs, (PA) up-regulated the mRNA level of NOX1, but not of NOX2 or NOX4. The production of nitric oxide by LSECs was significantly attenuated by PA-treatment in WT but not in Nox1KO. When the in vitro relaxation of TWNT1, a cell line that originated from hepatic stellate cells, was assessed by the gel contraction assay, the relaxation of stellate cells induced by LSECs was attenuated by PA treatment. In contrast, the relaxation effect of LSECs was preserved in cells isolated from Nox1KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impaired hepatic microcirculation through the reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the progression of NAFLD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: oxygen

Metabolic regulation of NLRP3.

A shift in our understanding of macrophage biology has come about as a result of recent discoveries in the area of metabolic reprogramming of macrophages. The NLRP3 inflammasome drives the activation of caspase-1, leading to the production of IL-1β, IL-18, and a type of cell death termed pyroptosis. The NLRP3 inflammasome has been shown to sense metabolites such as palmitate, uric , and cholesterol crystals and is inhibited by ketone bodies produced during metabolic flux. The NLRP3 inflammasome has also been shown to be regulated by mitochondrial reactive species and components of glycolysis, such as Hexokinase. Here, we review these findings and discuss their importance for inflammation and furthermore discuss potential therapeutic benefits of targeting NLRP3.© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: oxygen

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: oxygen

Metabolic dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the energy source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal lipid loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different metabolic conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: oxygen

EndoC-βH1 cells display increased sensitivity to sodium palmitate when cultured in DMEM/F12 medium.

Aims - Human pancreatic islets are known to die in response to the free fatty of sodium palmitate when cultured in vitro. This is in contrast to EndoC-βH1 cells, which in our hands are not sensitive to the cell death-inducing effects sodium palmitate, making these cells seemingly unsuitable for lipotoxicity studies. However, the EndoC-βH1 cells are routinely cultured in a nutrient mixture based on Dulbecco\'s Modified Eagle Medium (DMEM), which may not be the optimal choice for studies dealing with lipotoxicity. The aim of the present investigation was to define culture conditions that render EndoC-βH1 cells sensitive to toxic effects of sodium palmitate. Methods - EndoC-βH1 cells were cultured at standard conditions in either DMEM or DMEM/F12 culture medium. Cell death was analyzed using propidium iodide staining and flow cytometry. Insulin release and content was quantified using a human insulin ELISA. Results - We presently observe that substitution of DMEM for a DMEM/Ham\'s F12 mixture (50%/50% vol/vol) renders the cells sensitive to the apoptotic effects of sodium palmitate and sodium palmitate + high glucose leading to an increased cell death. Supplementation of the DMEM culture medium with linoleic partially mimicked the effect of DMEM/F12. Culture of EndoC-βH1 cells in DMEM/F12 resulted also in increased proliferation, ROS production and insulin contents, but markers for metabolic stress, autophagy or amyloid deposits were unaffected. Conclusions - The culture conditions for EndoC-βH1 cells can be modified so these cells display signs of lipotoxicity in response to sodium palmitate.

Keyword: oxygen

The Glucotoxicity Protecting Effect of Ezetimibe in Pancreatic Beta Cells via Inhibition of CD36.

Inhibition of CD36, a fatty transporter, has been reported to prevent glucotoxicity and ameliorate high glucose induced beta cell dysfunction. Ezetimibe is a selective cholesterol absorption inhibitor that blocks Niemann Pick C1-like 1 protein, but may exert its effect through suppression of CD36. We attempted to clarify the beneficial effect of ezetimibe on insulin secreting cells and to determine whether this effect is related to change of CD36 expression. mRNA expression of insulin and CD36, intracellular peroxide level and glucose stimulated insulin secretion (GSIS) under normal (5.6 mM) or high glucose (30 mM) condition in INS-1 cells and primary rat islet cells were compared. Changes of the aforementioned factors with treatment with ezetimibe (20 μM) under normal or high glucose condition were also assessed. mRNA expression of insulin was decreased with high glucose, which was reversed by ezetimibe in both INS-1 cells and primary rat islets. CD36 mRNA expression was increased with high glucose, but decreased by ezetimibe in INS-1 cells and primary rat islets. Three-day treatment with high glucose resulted in an increase in intracellular peroxide level; however, it was decreased by treatment with ezetimibe. Decrease in GSIS by three-day treatment with high glucose was reversed by ezetimibe. Palmitate uptake following exposure to high glucose conditions for three days was significantly elevated, which was reversed by ezetimibe in INS-1 cells. Ezetimibe may prevent glucotoxicity in pancreatic β-cells through a decrease in fatty influx via inhibition of CD36.

Keyword: oxygen

The Ethanol Extract of Larvae, Containing Fatty acids and Amino acids, Exerts Anti-Asthmatic Effects through Inhibition of the GATA-3/Th2 Signaling Pathway in Asthmatic Mice.

larvae (HD), a natural product from an insect resource, possesses many pharmacological properties, including anticoagulant, antitumor, anti-inflammatory, and analgesic activity. The major bioactive ingredients include oleic , , palmitoleic , linoleic , proline, and glutamic . Although HD is associated with immunoregulatory activities in allergic diseases, the therapeutic mechanisms of the action of HD in allergic diseases have not been investigated. The aim of this study was to evaluate the anti-asthmatic potential of HD in an ovalbumin (OVA)-induced mouse model of allergic asthma. Moreover, the anti-inflammatory potential of HD was examined to identify a plausible mechanism of action of HD in vitro. HD strongly reduced goblet cell hyperplasia, eosinophil infiltration, and reactive species (ROS), which reduced airway hyperresponsiveness (AHR), inflammation, and the expression of Th2 cytokines (IL-5 and IL-13) in bronchoalveolar lavage fluid (BALF). The expression of IL-5, IL-4, eotaxin-2, lysyl oxidase-like 2 (loxl2), and GATA-binding protein 3 (GATA-3) was attenuated in the lungs. In an in vitro assay, HD exerted immunomodulatory effects through the suppression of Th2 cytokines (IL-5, IL-13), IL-17, and tumor necrosis factor (TNF)-α production through downregulation of GATA-3 expression in EL-4 T cells. These findings suggest that the anti-asthmatic activity of HD may occur through the suppression of Th2 cytokines and total Immunoglobulin E (IgE) production by inhibition of the GATA-3 transcription pathway. Our results suggest that HD may be a potential alternative therapy, or a novel therapeutic traditional medicine, for the treatment of allergic asthma.

Keyword: oxygen

Palmitate-induced impairment of glucose-stimulated insulin secretion precedes mitochondrial dysfunction in mouse pancreatic islets.

It has been well established that excessive levels of glucose and palmitate lower glucose-stimulated insulin secretion (GSIS) by pancreatic β-cells. This β-cell \'glucolipotoxicity\' is possibly mediated by mitochondrial dysfunction, but involvement of bioenergetic failure in the pathological mechanism is the subject of ongoing debate. We show in the present study that increased palmitate levels impair GSIS before altering mitochondrial function. We demonstrate that GSIS defects arise from increased insulin release under basal conditions in addition to decreased insulin secretion under glucose-stimulatory conditions. Real-time respiratory analysis of intact mouse pancreatic islets reveals that mitochondrial ATP synthesis is not involved in the mechanism by which basal insulin is elevated. Equally, mitochondrial lipid oxidation and production of reactive species (ROS) do not contribute to increased basal insulin secretion. Palmitate does not affect KCl-induced insulin release at a basal or stimulatory glucose level, but elevated basal insulin release is attenuated by palmitoleate and associates with increased intracellular calcium. These findings deepen our understanding of β-cell glucolipotoxicity and reveal that palmitate-induced GSIS impairment is disconnected from mitochondrial dysfunction, a notion that is important when targeting β-cells for the treatment of diabetes and when assessing islet function in human transplants.© 2016 Authors; published by Portland Press Limited.

Keyword: oxygen

stimulates interleukin-8 via the TLR4/NF-κB/ROS pathway and induces mitochondrial dysfunction in bovine oviduct epithelial cells.

We investigated the effect of (PA), a major saturated fatty in NEFA, on bovine oviduct epithelial cells (OECs) during in vitro cell culture.Bovine oviductal tissues ipsilateral to the corpus luteum were collected 1-3\xa0days after ovulation; the OECs were isolated and cultured.PA increased lipid accumulation and activated caspase-3 in OECs, resulting in decreased cell proliferation. PA also stimulated the secretion of inflammatory cytokine interleukin (IL)-8 depending on TLR4, NF-κB activation, and reactive species (ROS) production. Moreover, PA induced mitochondrial dysfunction, including mitochondrial fission, ATP production, and mitochondrial ROS production. It also increased levels of LC3 and p62 proteins, suggesting autophagy induction in OECs.We suggest that bovine OECs recognize an excessive increase in endogenous and sterile danger signals, such as PA, which may contribute to chronic oviductal inflammation, resulting in infertility associated with oviductal dysfunction.© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: oxygen

Effects of saturated and omega-3 polyunsaturated fatty acids on Sertoli cell apoptosis.

Obesity is believed to negatively affect male semen quality and is accompanied by dysregulation of free fatty (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report obesity decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive species were elevated. Furthermore, we demonstrated by in vitro assays that saturated (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated fatty acids might be the cause of obesity-induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs.FFA: free fatty ; HFD: high-fat diet; SD: standard diet; PA: ; PUFA: polyunsaturated fatty ; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic fatty liver disease; DCFH-DA: 2\', 7\' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic ; PI: propidium iodide; DHA: docosahexenoic .

Keyword: oxygen

Protective role of the novel hybrid 3,5-dipalmitoyl-nifedipine in a cardiomyoblast culture subjected to simulated ischemia/reperfusion.

This work investigated the acute effects of the calcium channel blocker nifedipine and its new fatty hybrid derived from , 3,5-dipalmitoyl-nifedipine, compared to endocannabinoid anandamide during the process of inducing ischemia and reperfusion in cardiomyoblast H9c2 heart cells. The cardiomyoblasts were treated in 24 or 96-well plates (according to the test being performed) and maintaining the treatment until the end of hypoxia induction. The molecules were tested at concentrations of 10 and 100μM, cells were treated 24h after assembling the experimental plates and immediately before the I/R. Cell viability, apoptosis and necrosis, and generation of reactive species were evaluated. Nifedipine and 3,5-dipalmitoyl-nifedipine were used to assess radical scavenging potential and metal chelation. All tested molecules managed to reduce the levels of reactive species compared to the starvation+vehicle group. In in vitro assays, 3,5-dipalmitoyl-nifedipine showed more antioxidant activity than nifedipine. These results indicate the ability of this molecule to act as a powerful ROS scavenger. Cell viability was highest when cells were induced to I/R by both concentrations of anandamide and the higher concentration of DPN. These treatments also reduced cell death. Therefore, it was demonstrated that the process of hybridization of nifedipine with two chains assigns a greater cardioprotective effect to this molecule, thereby reducing the damage caused by hypoxia and reoxygenation in cardiomyoblast cultures.Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Keyword: oxygen

Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis.

Mitochondrial dysfunction plays a crucial role in the development of non-alcoholic steatohepatitis (NASH). However, the regulator of mitochondrial dysfunction in the pathogenesis of NASH is still largely unclear. CXCR3 is an essential pro-inflammatory factor in chronic liver diseases. We explored the significance of CXCR3 in regulating mitochondrial function during NASH development in animal models and cultured hepatocytes.The effects of CXCR3 on mitochondrial function were evaluated by genetic knockout or pharmacological inhibition in mouse models and . The ultrastructural changes of mitochondria were assessed by transmission electron microscopy (TEM). Hepatic levels of mitochondrial reactive species (ROS), DNA damage, membrane potential and ATP were examined.CXCR3 ablation by genetic knockout or pharmacological inhibition in mice protected against NASH development by influencing mitochondrial function. Similarly, depletion of CXCR3 reduced steatohepatitis injury in cultured hepatocytes. TEM analysis revealed that liver mitochondrial integrity was much improved in CXCR3 knockout (CXCR3) compared to wildtype (WT) mice. In agreement with this, impaired mitochondrial function was pronounced in WT mice compared to CXCR3 mice, evidenced by increased protein expression of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and decreased protein expression of mitofusin-1 (MFN1). Mitochondrial dysfunction was induced in AML-12 hepatocytes by methionine and choline deficient medium and in HepG2 cells by . The impaired mitochondrial function in both cell lines was evidenced by reduced membrane potential and ATP content, and by increased mitochondrial ROS accumulation and DNA damage. However, CXCR3 knockdown by siCXCR3 significantly diminished the mitochondrial dysfunction in both AML-12 and HepG2 hepatocytes. In addition, inhibition of CXCR3 by CXCR3 specific antagonists SCH546738 and AMG487 restored mitochondrial function and inhibited mitochondrial-dependent apoptosis in the liver of WT mice fed with methionine and choline deficient diet.CXCR3 induces mitochondrial dysfunction, which contributes to the pathogenesis of steatohepatitis. Pharmacologic blockade of CXCR3 prevents mitochondrial dysfunction and restores the severity of steatohepatitis, indicating a potential clinical impact for controlling the disease.

Keyword: oxygen

increases invasiveness of pancreatic cancer cells AsPC-1 through TLR4/ROS/NF-κB/MMP-9 signaling pathway.

Pancreatic cancer (PC) is an aggressive malady with proclivity for early metastasis. Overexpression of toll-like receptor 4 (TLR4) in pancreatic ductal adenocarcinoma, the most common type of pancreatic malignancy, correlates to tumor size, lymph node involvement, venous invasion and pathological stage. Lipopolysaccharides (LPS) are natural TLR4 ligands that have been shown to increase the invasive ability of PC cells. However, rapid inactivation of circulating LPS and low systemic absorption of inhaled LPS from the bronchoalveolar compartment make other agonists such as saturated fatty acids more suitable to be considered for TLR4-related cell invasiveness. Interestingly, PC risk was strongly associated to intake of saturated fat from animal food sources, in particular to consumption of saturated (PA). In the present study, we investigated the influence of PA on the invasive capacity of human PC cells AsPC-1. Using specific inhibitors, we found that PA stimulation of these tumor cells induced a TLR4-mediated cell invasion. Our results also indicate that the signaling events downstream of TLR4 involved generation of reactive species, activation of nuclear factor-kappa beta, and secretion and activation of matrix metalloproteinase 9 (MMP-9). Furthermore, PA stimulation decreased the levels of the micro RNA 29c (miR-29c). Of note, while inhibition of miR-29c increased MMP-9 mRNA levels, MMP-9 secretion and activation, and invasiveness, miR-29c mimic abrogated all these PA-stimulated effects. These results strongly suggest that miR-29c could be an attractive potential pharmacological agent for antitumoral therapy in PC.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: oxygen

Alpha-linolenic protects the developmental capacity of bovine cumulus-oocyte complexes matured under lipotoxic conditions in vitro.

Elevated concentrations of free fatty acids (FFAs), predominantly , stearic, and oleic acids (PSO), exert detrimental effects on oocyte developmental competence. This study examined the effects of omega-3 alpha-linolenic (ALA) during in vitro oocyte maturation (IVM) in the presence of PSO on subsequent embryo development and quality, and the cellular mechanisms that might be involved. Bovine cumulus-oocyte complexes (COCs) were supplemented during IVM with ALA (50 μM), PSO (425 μM), or PSO+ALA. Compared with FFA-free controls (P\xa0<\xa00.05), PSO increased embryo fragmentation and decreased good quality embryos on day 2 postfertilization. Day 7 blastocyst rate was also reduced. Day 8 blastocysts had lower cell counts and higher apoptosis but normal metabolic profile. In the PSO group, cumulus cell (CC) expansion was inhibited with an increased CC apoptosis while COC metabolism was not affected. Mitochondrial inner membrane potential (MMP; JC-1 staining) was reduced in the CCs and oocytes. Heat shock protein 70 (HSP70) but not glucose-regulated protein 78 kDa (GRP78, known as BiP; an endoplasmic reticulum stress marker) was upregulated in the CCs. Higher reactive species levels (DCHFDA staining) were detected in the oocytes. In contrast, adding ALA in the presence of PSO normalized embryo fragmentation, cleavage, blastocyst rates, and blastocyst quality compared to controls (P\xa0>\xa00.05). Combined treatment with ALA also reduced CC apoptosis, partially recovered CC expansion, abrogated the reduction in MMP in the CCs but not in the oocytes, and reduced BiP and HSP70 expression in CCs, compared with PSO only (P\xa0<\xa00.05). In conclusion, ALA supplementation protected oocyte developmental capacity under lipotoxic conditions mainly by protecting cumulus cell viability.© The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: oxygen

Sumoylation of PPARγ contributes to vascular endothelium insulin resistance through stabilizing the PPARγ-NcoR complex.

Sumoylation of peroxisome proliferator-activated receptor\xa0γ (PPARγ) affects its stabilization, sublocalization, and transcriptional activity. However, it remains largely unknown whether PPARγ sumoylation inhibits the transactivation effect, leading to endothelium insulin resistance (IR). To test this possibility, human umbilical vascular endothelial cells (HUVECs) with a 90% confluence were randomly allocated to two batches. One batch was first pretreated with or without vitamin E for 24\u2009hr and the other infected with adenoviruses containing either PIAS1-shRNA (protein inhibitor of activated STAT1-short hairpin RNA) or scramble shRNA. Cells were suffered from high glucose and (PA) exposure for further 48\u2009hr. The levels of PPARγ, p-IKK, IKK, and NcoR (nuclear corepressors) were measured by western blot analysis. The interaction of IKK and PIAS1, as well as the PPARγ sumoylation, were examined by coimmunoprecipitation. The results showed that the exposure of high glucose and PA\xa0induced reactive species (ROS) production and IKK activation in HUVECs, promoting the interaction of IKK and PIAS1 and the sumoylation of PPARγ. However, vitamin E and PIAS1-shRNA partially decreased ROS production and IKK activation induced by high glucose and PA exposure. These data indicate that ROS-IKK-PIAS1 pathway mediates PPARγ sumoylation, leading to endothelium IR via stabilizing PPARγ-NcoR complex. These findings benefit understanding of regulatory networks of insulin signaling, which might provide a potential target to prevent and cure IR-related diseases.© 2019 Wiley Periodicals, Inc.

Keyword: oxygen

Long-chain fatty activates hepatocytes through CD36 mediated oxidative stress.

Accumulating evidence suggests that activated hepatocytes are involved in the deposition of the excess extracellular matrix during liver fibrosis via the epithelial to mesenchymal transition. Lipid accumulation in hepatocytes are implicated in the pathogenesis of chronic liver injury. CD36 is known to mediate long-chain fatty (LCFA) uptake and lipid metabolism. However, it is unclear whether LCFA directly promotes hepatocyte activation and the involved mechanisms have not been fully clarified.Mice were fed with a high fat diet (HFD) and normal hepatocyte cells (Chang liver cells) were treated with (PA) in vivo and in vitro. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to examine the gene and protein expression of molecules involved in hepatic fibrogenesis and hepatocyte activation. CD36 was knocked down by transfecting CD36 siRNA into hepatocyte cells. Hydrogen peroxide (HO) and reactive species (ROS) levels were detected using commercial kits.HFD induced a profibrogenic response and up-regulated CD36 expression in vivo. Analogously, PA increased lipid accumulation and induced human hepatocyte activation in vitro, which was also accompanied by increased CD36 expression. Interestingly, knockdown of CD36 resulted in a reduction of hepatocyte lipid deposition and decreased expression of Acta2 (34% decrease), Vimentin (29% decrease), Desmin (60% decrease), and TGF-β signaling pathway related genes. In addition, HFD and PA increased the production of HO in vivo (48% increase) and in vitro (385% increase), and the antioxidant, NAC, ameliorated PA-induced hepatocyte activation. Furthermore, silencing of CD36 in vitro markedly attenuated PA-induced oxidative stress (HO: 41% decrease; ROS: 39% decrease), and the anti-activation effects of CD36 knockdown could be abolished by pretreatment with HO.Our study demonstrated that LCFA facilitates hepatocyte activation by up-regulating oxidative stress through CD36, which could be an important mechanism in the development of hepatic fibrosis.

Keyword: oxygen

Telmisartan protects against high glucose/high lipid-induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: oxygen

Adelmidrol + sodium hyaluronate in IC/BPS or conditions associated to chronic urothelial inflammation. A translational study.

Interstitial cystitis/painful bladder syndrome (IC/PBS) is a chronic bladder condition characterized by frequent urination, bladder inflammation and pain. It is a particular challenging disease and a clear unmet medical need in terms of identifying new therapeutic strategies. The aim of study was to evaluate the anti-inflammatory effects of intravesical Vessilen (a new formulation of 2% adelmidrol (the diethanolamide derivative of azelaic ) + 0.1% sodium hyaluronate) administration in rodent models of IC/BPS and in IC/BPS patients or other bladder disorders. Acute and chronic animal models of cystitis were induced by a single or repetitive intraperitoneal injections of cyclophosphamide (CYP); patients with IC/BPS or with bladder pain syndrome associated with symptoms of the lower urinary tract treated once weekly by bladder instillation of Vessilen for 8 weeks. CYP instillation caused macroscopic and histological bladder alterations, inflammatory infiltrates, increased mast cell numbers, bladder pain, increased expression of nitrotyrosine, decreased expression of endothelial tight junction zonula occludens-1. Intravesical Vessilen® treatment was able to ameliorate CYP induced bladder inflammation and pain by inhibiting nuclear factor-κB pathway and inflammatory mediator levels as well as reduced mechanical allodynia and nerve growth factor levels. A significant improvement in quality of life and symptom intensity were evident in patients with IC/BPS or other bladder disorders treated with Vessilen. Vessilen could be a new therapeutic approach for human cystitis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

New fatty dihydropyridines present cardioprotective potential in H9c2 cardioblasts submitted to simulated ischemia and reperfusion.

Nifedipine is a calcium channel blocker dihydropyridine that has been used in the treatment of hypertension. The production of reactive species and calcium overload are the main contributors to myocardial ischemia-reperfusion (I / R) injury. We investigated the ability of novel dihydropyridines (DHPs) to improve the effect of protecting against the injury induced by ischemia and reperfusion in cardioblasts when compared to nifedipine. Forty three DHPs were created varying the fatty chains derived from , stearic and oleic acids and aromatic moiety in addition to the addition of chemical elements such as chlorine, nitrogen dioxide, furfural, hydroxyl and methoxy. Cytotoxicity and inhibition of linoleic oxidation were evaluated for all new DHPs and also for nifedipine. The alpha-tocopherol and butylated hydroxytoluene (BHT) were used as antioxidants controls. The compounds with the best antioxidant potential were used in the ischemia and reperfusion (I / R) induction test in cardioblasts (H9c2). Cardioblasts were treated 24\u2009h after assembly of plates and submitted to the ischemia simulation (30\u2009min), after which, normoxia and cellular nutrition conditions were reestablished, simulating reperfusion (additional 30\u2009min). Right after, cell viability, apoptosis, necrosis, and the generation of reactive species (ROS) were evaluated. Cell viability during I / R was not altered in cells treated with nifedipine, BHT and the new DHP composed of with hydroxyl group in the aromatic substituent. The other new DHPs increased cell viability during I / R simulation and reduced levels of reactive species compared to the I / R group, demonstrating the antioxidant capacity of the new DHPs. Therefore, DHPS with and oleic acids in the C3 and C5 position with NO2 or Cl in aromatic moiety, presented the highest antioxidant potential (linoleic oxidant test). The new DHPs increased cell viability during I / R simulation and reduced levels of reactive species compared to the ischemia and reperfusion group, demonstrating the antioxidant capacity of the new DHPs. Taken together, these results indicate that those new DHPs have a greater cardioprotective antioxidant capacity to face the damages of ischemia and reperfusion.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: oxygen

stimulates energy metabolism and inhibits insulin/PI3K/AKT signaling in differentiated human neuroblastoma cells: The role of mTOR activation and mitochondrial ROS production.

The high consumption of saturated lipids has been largely associated with the increasing prevalence of metabolic diseases. In particular, saturated fatty acids such as (PA) have been implicated in the development of insulin resistance in peripheral tissues. However, how neurons develop insulin resistance in response to lipid overload is not fully understood. Here, we used cultured rat cortical neurons and differentiated human neuroblastoma cells to demonstrate that PA blocks insulin-induced metabolic activation, inhibits the activation of the insulin/PI3K/Akt pathway and activates mTOR kinase downstream of Akt. Despite the fact that fatty acids are not normally used as a significant source of fuel by neural cells, we also found that short-term neuronal exposure to PA reduces the NAD/NADH ratio, indicating that PA modifies the neuronal energy balance. Finally, inhibiting mitochondrial ROS production with mitoTEMPO prevented the deleterious effect of PA on insulin signaling. This work provides novel evidence of the mechanisms behind saturated fatty -induced insulin resistance and its metabolic consequences on neuronal cells.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Systematic evaluation of phenolic compounds and protective capacity of a new mulberry cultivar J33 against -induced lipotoxicity using a simulated digestion method.

This research aimed to investigate the protective effects of a new mulberry cultivar J33 with simulated gastrointestinal digestion against (PA)-induced lipotoxicity. LC-MS analysis revealed that the contents of four flavonoid glycosides (quercetin rhamnosylhexoside hexoside, quercetin rhamnosylhexoside, quercetin hexoside, kaempferol rhamnosylhexoside) increased after digestion. Besides, mulberry digest (MBD) at 0.5-2\u202fmg/mL significantly reduced PA-induced lipotoxicity in human hepatocytes, while mulberry extract without digestion (MBE) showed no protection. Further investigations demonstrated that the protection of MBD was attributed to two aspects. On the one hand, MBD could attenuate PA-induced oxidative stress by suppressing ROS accumulation, regulating intracellular glutathione and ameliorating mitochondrial dysfunction. On the other hand, MBD could promote PA incorporation into inert triglycerides (TG) to deal with the acute lipid overload, reducing the lipotoxicity caused by PA. Overall, our research might provide a new perspective of mulberry cultivar J33 in ameliorating non-alcoholic fatty liver disease (NAFLD).Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Let there be light: stability of monolayers at the air/salt water interface in the presence and absence of simulated solar light and a photosensitizer.

Long-chain fatty monolayers are known surfactants present at air/water interfaces. However, little is known about the stability of these long-chain fatty monolayers in the presence of solar radiation. Here we have investigated, for the first time, the stability of monolayers on salt water interfaces in the presence and absence of simulated solar light with and without a photosensitizer in the underlying salt subphase. Using surface sensitive probes to measure changes in the properties of these monolayers upon irradiation, we found that the monolayers become less stable in the presence of light and a photosensitizer, in this case humic , in the salt solution. The presence of the photosensitizer is essential in significantly reducing the stability of the monolayer upon irradiation. The mechanism for this loss of stability is due to interfacial photochemistry involving electronically excited humic and molecular reacting with at the interface to form more oxygenated and less surface-active species. These oxygenated species can then more readily partition into the underlying solution.

Keyword: oxygen

Oriented surface epitope imprinted polymer-based quartz crystal microbalance sensor for cytochrome c.

A quartz crystal microbalance (QCM) sensor for detecting cytochrome c based on an oriented surface epitope imprinted polymer was fabricated in this paper. By using the -modified epitope of cytochrome c as the template and the 3-aminopropyltriethoxysilane as the monomer, we prepared a new oriented surface epitope imprinted polymer by the reverse microemulsion polymerization. The prepared oriented imprinted polymer had better imprinting effect than the non-oriented imprinted polymer. And compared to previous studies, this polymerization method is simple and could be carried out at room temperature in the presence of , under regular atmospheric conditions. Then, by combining the advantages of molecularly imprinted polymers and QCM sensors, we used the prepared polymer to establish a QCM sensor. The described sensor showed good sensitivity and selectivity towards cytochrome c. The linear range was from 0.005\u202fμg\u202fmL to 0.050\u202fμg\u202fmL and the detection limit was 3.6\u202fng\u202fmL which is lower than most of previous works. Besides, it could be used for real sample analysis and had satisfactory reproducibility and accuracy. This work proposed a new way of fabricating oriented surface epitope imprinted polymers-based QCM sensors for selectively detecting proteins at very low concentrations.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: oxygen

Adiponectin protects induced endothelial inflammation and insulin resistance via regulating ROS/IKKβ pathways.

Endothelial inflammation and insulin resistance (IR) has been closely associated with endothelial dysfunction. Adiponectin (APN), an adipocyte-secreted hormone from adipose tissues, showed cardioprotective effects. Here, the protective effect of APN on (PA)-induced endothelial inflammation and IR was investigated. Cultured human umbilical vein endothelial cells (HUVECs) were treated with PA without or without APN pretreatment. The expression of inflammatory cytokines TNF-α, IL-6, adhesion molecule ICAM-1 were determined by western blotting, ELISA, and real-time PCR. The protein expression and protein-protein interaction were determined by western blotting and immunoprecipitation. The intracellular reactive species (ROS) and nitric oxide (NO) production were monitored with fluorescence probes. PA-induced secretion of TNF-α, IL-6, and expression of ICAM-1 at protein and mRNA levels, which was significantly inhibited by APN. PA treatment caused increase of ROS generation, NOX2, p-IKKβ, p-IκBα, p-p65 expression, and p-IκBα-IKKβ interaction, which were all partly reversed by APN. ROS scavenger N-acetylcysteine (NAC) and NF-κB inhibitor PDTC showed similar effect on PA-induced secretion of TNF-α, IL-6, and expression of ICAM-1. Furthermore, APN and NAC pretreatment restored PA-induced increase of p-IRS-1(S307), decrease of p-IRS-1(Tyr). In addition, insulin-triggered expression of p-IRS-1(Tyr), p-PI3K, p-AKT, p-eNOS and NO generation were inhibited by PA, which were also restored by both APN and NAC. These results suggested that APN ameliorated endothelial inflammation and IR through ROS/IKKβ pathway. This study shed new insights into the mechanisms of APN\'s cardiovascular protective effect.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Wax Ester Fermentation and Its Application for Biofuel Production.

In Euglena cells under anaerobic conditions, paramylon, the storage polysaccharide, is promptly degraded and converted to wax esters. The wax esters synthesized are composed of saturated fatty acids and alcohols with chain lengths of 10-18, and the major constituents are myristic and myristyl alcohol. Since the anaerobic cells gain ATP through the conversion of paramylon to wax esters, the phenomenon is named "wax ester fermentation". The wax ester fermentation is quite unique in that the end products, i.e. wax esters, have relatively high molecular weights, are insoluble in water, and accumulate in the cells, in contrast to the common fermentation end products such as lactic and ethanol.A unique metabolic pathway involved in the wax ester fermentation is the mitochondrial fatty synthetic system. In this system, fatty are synthesized by the reversal of β-oxidation with an exception that trans-2-enoyl-CoA reductase functions instead of acyl-CoA dehydrogenase. Therefore, acetyl-CoA is directly used as a C donor in this fatty synthesis, and the conversion of acetyl-CoA to malonyl-CoA, which requires ATP, is not necessary. Consequently, the mitochondrial fatty synthetic system makes possible the net gain of ATP through the synthesis of wax esters from paramylon. In addition, acetyl-CoA is provided in the anaerobic cells from pyruvate by the action of a unique enzyme, sensitive pyruvate:NADP oxidoreductase, instead of the common pyruvate dehydrogenase multienzyme complex.Wax esters produced by anaerobic Euglena are promising biofuels because myristic (C) in contrast to other algal produced fatty acids, such as (C) and stearic (C), has a low freezing point making it suitable as a drop-in jet fuel. To improve wax ester production, the molecular mechanisms by which wax ester fermentation is regulated in response to aerobic and anaerobic conditions have been gradually elucidated by identifying individual genes related to the wax ester fermentation metabolic pathway and by comprehensive gene/protein expression analysis. In addition, expression of the cyanobacterial Calvin cycle fructose-1,6-bisphosphatase/sedohepturose-1,7-bisphosphatase, in Euglena provided photosynthesis resulting in increased paramylon accumulation enhancing wax ester production. This chapter will discuss the biochemistry of the wax ester fermentation, recent advances in our understanding of the regulation of the wax ester fermentation and genetic engineering approaches to increase production of wax esters for biofuels.

Keyword: oxygen

Suppression of Lipogenesis via Reactive Species-AMPK Signaling for Treating Malignant and Proliferative Diseases.

Systemic diseases often have common characteristics. The aim of this study was to investigate the feasibility of targeting common pathological metabolism to inhibit the progression of malignant and proliferative diseases.Gefitinib-resistant (G-R) nonsmall-cell lung cancer (NSCLC) and rheumatoid arthritis (RA) were studied as conditions representative of malignant and proliferative diseases, respectively. Strong lipogenic activity and high expression of sterol regulatory element-binding protein 1 (SREBP1) were found in both G-R NSCLC cells and synovial fibroblasts from RA patients (RASFs). Berberine (BBR), an effective suppressor of SREBP1 and lipogenesis regulated through reactive species (ROS)/AMPK pathway, selectively inhibited the growth of G-R NSCLC cells and RASFs but not that of normal cells. It effectively caused mitochondrial dysfunction, activated ROS/AMPK pathway, and finally suppressed cellular lipogenesis and cell proliferation. Addition of ROS blocker, AMPK inhibitor, and significantly reduced the effect of BBR. In an in vivo study, treatment of BBR led to significant inhibition of mouse tumor xenograft growth and remarkably slowed down the development of adjuvant-induced arthritis in rats. Innovation and Conclusion: Targeting ROS/AMPK/lipogenesis signaling pathway selectively inhibited the growth of G-R NSCLC cells and the progress of RASFs in vitro and in vivo, which provides a new avenue for treating malignancies and proliferative diseases. Antioxid. Redox Signal. 28, 339-357.

Keyword: oxygen

Bi-allelic mutations of LONP1 encoding the mitochondrial LonP1 protease cause pyruvate dehydrogenase deficiency and profound neurodegeneration with progressive cerebellar atrophy.

LonP1 is crucial for maintaining mitochondrial proteostasis and mitigating cell stress. We identified a novel homozygous missense LONP1 variant, c.2282 C\xa0>\xa0T, (p.Pro761Leu), by whole-exome and Sanger sequencing in two siblings born to healthy consanguineous parents. Both siblings presented with stepwise regression during infancy, profound hypotonia and muscle weakness, severe intellectual disability and progressive cerebellar atrophy on brain imaging. Muscle biopsy revealed the absence of ragged-red fibers, however, scattered cytochrome c oxidase-negative staining and electron dense mitochondrial inclusions were observed. Primary cultured fibroblasts from the siblings showed normal levels of mtDNA and mitochondrial transcripts, and normal activities of oxidative phosphorylation complexes I through V. Interestingly, fibroblasts of both siblings showed glucose-repressed consumption compared to their mother, whereas galactose and utilization were similar. Notably, the siblings\' fibroblasts had reduced pyruvate dehydrogenase (PDH) activity and elevated intracellular lactate:pyruvate ratios, whereas plasma ratios were normal. We demonstrated that in the siblings\' fibroblasts, PDH dysfunction was caused by increased levels of the phosphorylated E1α subunit of PDH, which inhibits enzyme activity. Blocking E1α phosphorylation activated PDH and reduced intracellular lactate concentrations. In addition, overexpressing wild-type LonP1 in the siblings\' fibroblasts down-regulated phosphoE1α. Furthermore, in vitro studies demonstrated that purified LonP1-P761L failed to degrade phosphorylated E1α, in contrast to wild-type LonP1. We propose a novel mechanism whereby homozygous expression of the LonP1-P761L variant leads to PDH deficiency and energy metabolism dysfunction, which promotes severe neurologic impairment and neurodegeneration.

Keyword: oxygen

Moderate Exercise Prevents Functional Remodeling of the Anterior Pituitary Gland in Diet-Induced Insulin Resistance in Rats: Role of Oxidative Stress and Autophagy.

A sustained elevation of glucocorticoid production, associated with the establishment of insulin resistance (IR) could add to the deleterious effects of the IR state. The aim of this study is to analyze the consequences of long-term feeding with a sucrose-rich diet (SRD) on Pomc/ACTH production, define the underlying cellular processes, and determine the effects of moderate exercise (ME) on these parameters. Animals fed a standard chow with or without 30% sucrose in the drinking water were subjected to ME. Circulating hormone levels were determined, and pituitary tissues were processed and analyzed by immunobloting and quantitative real-time PCR. Parameters of oxidative stress (OxS), endoplasmic reticulum stress, and autophagy were also determined. Rats fed SRD developed a decrease in pituitary Pomc/ACTH expression levels, increased expression of antioxidant enzymes, and induction of endoplasmic reticulum stress and autophagy. ME prevented pituitary dysfunction as well as induction of antioxidant enzymes and autophagy. Reporter assays were performed in AtT-20 corticotroph cells incubated in the presence of . Pomc transcription was inhibited by -dependent induction of OxS and autophagy, as judged by the effect of activators and inhibitors of both processes. Long-term feeding with SRD triggers the generation of OxS and autophagy in the pituitary gland, which could lead to a decline in Pomc/ACTH/glucocorticoid production. These effects could be attributed to an increase in fatty acids availability to the pituitary gland. ME was able to prevent these alterations, suggesting additional beneficial effects of ME as a therapeutic strategy in the management of IR.

Keyword: oxygen

Curcumin Ester Facilitates Protection of Neuroblastoma against Oligomeric Aβ40 Insult.

The generation of reactive species (ROS) caused by amyloid-β (Aβ) is considered to be one of mechanisms underlying the development of Alzheimer\'s disease. Curcumin can attenuate Aβ-induced neurotoxicity through ROS scavenging, but the protective effect of intracellular curcumin on neurocyte membranes against extracellular Aβ may be compromised. To address this issue, we synthesized a curcumin ester (P-curcumin) which can be cultivated on the cell membrane and investigated the neuroprotective effect of P-curcumin and its interaction with Aβ.P-curcumin was prepared through chemical synthesis. Its structure was determined via nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). An MTT assay was used to assess Aβ cytotoxicity and the protective effect of P-curcumin on SH-SY5Y cells. The effect of P-curcumin on Aβ-induced ROS production in vitro and in vivo were assessed based on changes in dichlorofluorescein (DCF) fluorescence. A spectrophotometric method was employed to detect lipid peroxidation. To mimic the interaction of P-curcumin on cell membranes with Aβ, liposomes were prepared by thin film method. Finally, the interactions between free P-curcumin and P-curcumin cultivated on liposomes and Aβ were determined via spectrophotometry.A novel derivative, curcumin ester was prepared and characterized. This curcumin, cultivated on the membranes of neurocytes, may prevent Aβ-mediated ROS production and may inhibit the direct interaction between Aβ and the cellular membrane. Furthermore, P-curcumin could scavenge Aβ-mediated ROS as curcumin in vitro and in vivo, and had the potential to prevent lipid peroxidation. Morphological analyses showed that P-curcumin was better than curcumin at protecting cell shape. To examine P-curcumin\'s ability to attenuate direct interaction between Aβ and cell membranes, the binding affinity of Aβ to curcumin and P-curcumin was determined. The association constants for free P-curcumin and curcumin were 7.66 × 104 M-1 and 7.61 × 105 M-1, respectively. In the liposome-trapped state, the association constants were 3.71 × 105 M-1 for P-curcumin and 1.44× 106 M-1 for curcumin. With this data, the thermodynamic constants of P-curcumin association with soluble Aβ (ΔH, ΔS, and ΔG) were also determined.Cultivated curcumin weakened the direct interaction between Aβ and cell membranes and showed greater neuroprotective effects against Aβ insult than free curcumin.© 2017 The Author(s). Published by S. Karger AG, Basel.

Keyword: oxygen

Thioredoxin interacting protein mediates lipid-induced impairment of glucose uptake in skeletal muscle.

Insulin resistance (IR) is an important determinant of type-2 diabetes mellitus (T2DM). Free fatty acids (FFAs) induce IR by various mechanisms. A surfeit of circulating FFA leads to intra-myocellular lipid accumulation that induces mitochondrial ROS generation and worsens IR. However, the molecular mechanisms behind are unclear. We identified thioredoxin interacting protein (TxNIP), which is overexpressed in T2DM, to be a promoter of ROS-induced IR. We observed upregulation of TxNIP upon palmitate treatment in skeletal muscle cells that led to ROS generation and Glut-4 downregulation resulting in impaired glucose-uptake. FFA-induced overexpression of TxNIP gene was mediated through the activation of its bona-fide trans activator, ChREBP. Further, Palmitate-induced impairment in AMPK-SIRT-1 pathway resulted in overexpression of ChREBP. While Fenofibrate, abrogated PA-induced TxNIP expression and ROS generation in skeletal muscle cells, Saroglitazar, a dual PPARα/γ-agonist, not only inhibited PA-induced TXNIP expression but also led to greater improvement in glucose uptake. Taken together, TxNIP appears to be an important factor in FFA-induced ROS generation and IR in skeletal muscle cells, which can be modulated for the management of this complex disorder.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: oxygen

The effect of chronic exposure to high concentrations on the aerobic metabolism of human endothelial EA.hy926 cells.

A chronic elevation of circulating free fatty acids (FFAs) is associated with diseases like obesity or diabetes and can lead to lipotoxicity. The goals of this study were to assess the influence of chronic exposure to high (PAL) levels on mitochondrial respiratory functions in endothelial cells and isolated mitochondria. Human umbilical vein endothelial cells (EA.hy926 line) were grown for 6\xa0days in a medium containing either 100 or 150\xa0μM PAL. Growth at high PAL concentrations induced a considerable increase in fatty -supplied respiration and a reduction of mitochondrial respiration during carbohydrate and glutamine oxidation. High PAL levels elevated intracellular and mitochondrial superoxide generation; increased inflammation marker, acyl-coenzyme A (CoA) dehydrogenase, uncoupling protein 2 (UCP2), and superoxide dismutase 2 expression; and decreased hexokinase I and pyruvate dehydrogenase expression. No change in aerobic respiration capacity was observed, while fermentation was decreased. In mitochondria isolated from high PAL-treated cells, an increase in the oxidation of palmitoylcarnitine, a decrease in the oxidation of pyruvate, and an increase in UCP2 activity were observed. Our results demonstrate that exposure to high PAL levels induces a shift in endothelial aerobic metabolism toward the oxidation of fatty acids. Increased levels of PAL caused impairment and uncoupling of the mitochondrial oxidative phosphorylation system. Our data indicate that FFAs significantly affect endothelial oxidative metabolism, reactive species (ROS) formation, and cell viability and, thus, might contribute to endothelial and vascular dysfunction.

Keyword: oxygen

is a toll-like receptor 4 ligand that induces human dendritic cell secretion of IL-1β.

(PA) and other saturated fatty acids are known to stimulate pro-inflammatory responses in human immune cells via Toll-like receptor 4 (TLR4). However, the molecular mechanism responsible for fatty stimulation of TLR4 remains unknown. Here, we demonstrate that PA functions as a ligand for TLR4 on human monocyte derived dendritic cells (MoDCs). Hydrophobicity protein modeling indicated PA can associate with the hydrophobic binding pocket of TLR4 adaptor protein MD-2. Isothermal titration calorimetry quantified heat absorption that occurred during PA titration into TLR4/MD2, indicating that PA binds to TLR4/MD2. Treatment of human MoDCs with PA resulted in endocytosis of TLR4, further supporting the function of PA as a TLR4 agonist. In addition, PA stimulated DC maturation and activation based on the upregulation of DC costimulatory factors CD86 and CD83. Further experiments showed that PA induced TLR4 dependent secretion of the pro-inflammatory cytokine IL-1β. Lastly, our experimental data show that PA stimulation of NF-κB canonical pathway activation is regulated by TLR4 signaling and that reactive species may be important in upregulating this pro-inflammatory response. Our experiments demonstrate for the first time that PA activation of TLR4 occurs in response to direct molecular interactions between PA and MD-2. In summary, our findings suggest a likely molecular mechanism for PA induction of pro-inflammatory immune responses in human dendritic cells expressing TLR4.

Keyword: oxygen

Protocatechuic -Ameliorated Endothelial Oxidative Stress through Regulating Acetylation Level via CD36/AMPK Pathway.

As one of the main metabolites of anthocyanin, protocatechuic (PCA) possesses strong antioxidant activity. In the present study, we explored the capacity of PCA on the alleviation of endothelial oxidative stress and investigated the underlying mechanisms using RNA sequencing (RNA-Seq). In comparison with (PA)-treated cells, PCA (100 μM) significantly decreased the generations of 3-nitrotyrosine (3-NT) and 8-hydroxydeoxyguanosine (8-OHdG) (0.82 ± 0.01 vs 1.16 ± 0.05 and 0.80 ± 0.01 vs 1.48 ± 0.15, respectively, p < 0.01), two biomarkers of oxidative damage, and restored the levels of nitric oxide (NO) (0.97 ± 0.04 vs 0.54 ± 0.02, p < 0.01) and mitochondrial membrane potential (MMP) (0.96 ± 0.03 vs 0.86 ± 0.02, p < 0.01) in human umbilical vein endothelial cells (HUVECs). PCA also obviously reduced the level of reactive species (ROS) (0.86 ± 0.15 vs 2.67 ± 0.09, p < 0.01) in aorta from high-fat diet (HFD)-fed mice. RNA-Seq and Western blot analysis indicated that PCA markedly reduced the expression of cluster of differentiation 36 (CD36), a membrane fatty transporter, and reduced the generations of adenosine triphosphate (ATP) and acetyl coenzyme A (Ac-CoA). These effects of PCA were associated with decreased level of acetylated-lysine and restored the activity of manganese-dependent superoxide dismutase (MnSOD) through reducing the generation of Ac-CoA or activating Sirt1 and Sirt3 via a CD36/AMP-kinase (AMPK) dependent pathway.

Keyword: oxygen

Liraglutide ameliorates non-alcoholic steatohepatitis by inhibiting NLRP3 inflammasome and pyroptosis activation via mitophagy.

Non-alcoholic steatohepatitis (NASH) is a key step in the progression of non-alcoholic fatty liver disease (NAFLD), which causes serious health problems worldwide. The nucleotide-binding oligomerization domain, leucine-rich repeat-containing receptor-containing pyrin domain 3 (NLRP3) inflammasome and pyroptosis play crucial roles in the progression of NASH. Our team has provided clinical evidence of the effects of glucagon-like peptide-1 (GLP-1) on the improvement in liver function and histological resolution of NAFLD. Preliminary work has demonstrated that GLP-1 inhibited NLRP3 inflammasome activation in a mouse model of NAFLD. We further explored the potential molecular mechanisms underlying the anti-inflammatory effect of liraglutide, a long-acting GLP-1 analog, in the treatment of NASH. We established a HepG2 cell model of NASH using double stimulation with and lipopolysaccharide to assess NLRP3 inflammasome and pyroptotic cell activity and to evaluate mitochondrial function and mitophagy. Liraglutide reduced lipid accumulation, inhibited NLRP3 inflammasome and pyroptosis activation, attenuated mitochondrial dysfunction and reactive species generation, augmented mitophagy in hepatocytes. Mitophagy inhibition with 3-methyladenine/PINK1-directed siRNA weakened the liraglutide-mediated suppression of inflammatory injury. We propose that liraglutide suppresses NLRP3 inflammasome-induced hepatocyte pyroptosis via mitophagy to slow the progression of NASH.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: oxygen

Hydrogen Sulphide modulating mitochondrial morphology to promote mitophagy in endothelial cells under high-glucose and high-palmitate.

Endothelial cell dysfunction is one of the main reasons for type II diabetes vascular complications. Hydrogen sulphide (H S) has antioxidative effect, but its regulation on mitochondrial dynamics and mitophagy in aortic endothelial cells under hyperglycaemia and hyperlipidaemia is unclear. Rat aortic endothelial cells (RAECs) were treated with 40 mM glucose and 200 μM palmitate to imitate endothelium under hyperglycaemia and hyperlipidaemia, and 100 μM NaHS was used as an exogenous H S donor. Firstly, we demonstrated that high glucose and palmitate decreased H S production and CSE expression in RAECs. Then, the antioxidative effect of H S was proved in RAECs under high glucose and palmitate to reduce mitochondrial ROS level. We also showed that exogenous H S inhibited mitochondrial apoptosis in RAECs under high glucose and palmitate. Using Mito Tracker and transmission electron microscopy assay, we revealed that exogenous H S decreased mitochondrial fragments and significantly reduced the expression of p-Drp-1/Drp-1 and Fis1 compared to high-glucose and high-palmitate group, whereas it increased mitophagy by transmission electron microscopy assay. We demonstrated that exogenous H S facilitated Parkin recruited by PINK1 by immunoprecipitation and immunostaining assays and then ubiquitylated mitofusin 2 (Mfn2), which illuminated the mechanism of exogenous H S on mitophagy. Parkin siRNA suppressed the expression of Mfn2, Nix and LC3B, which revealed that it eliminated mitophagy. In summary, exogenous H S could protect RAECs against apoptosis under high glucose and palmitate by suppressing oxidative stress, decreasing mitochondrial fragments and promoting mitophagy. Based on these results, we proposed a new mechanism of H S on protecting endothelium, which might provide a new strategy for type II diabetes vascular complication.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: oxygen

Metabolic flexibility to lipid availability during exercise is enhanced in individuals with high insulin sensitivity.

Metabolic flexibility to lipid (MetFlex-lip) is the capacity to adapt lipid oxidation to lipid availability. Hypothetically, impaired MetFlex-lip in skeletal muscle induces accumulation of lipid metabolites that interfere with insulin signaling. Our aim was to compare MetFlex-lip during exercise in subjects with low (Low_IS) vs. high (High_IS) insulin sensitivity. Twenty healthy men were designated as Low_IS or High_IS on the basis of the median of the homeostatic model assessment of insulin resistance index. Groups had similar age, body mass index, and maximum uptake (V̇o). Subjects cycled at 50% V̇o until expending 650 kcal. Adaptation in lipid oxidation was calculated as the drop in respiratory quotient (RQ) at the end of exercise vs. the maximum RQ (ΔRQ). Lipid availability was calculated as the increase in circulating nonesterified fatty acids (NEFA) at the end of exercise vs. the minimum NEFA (ΔNEFA). ΔRQ as a function of ΔNEFA was used to determine MetFlex-lip. On average, RQ and circulating NEFA changed similarly in both groups. However, ΔRQ correlated with ΔNEFA in High_IS ( r\u2009=\u2009-0.83, P < 0.01) but not in Low_IS ( r\u2009=\u2009-0.25, P = 0.48) subjects. Thus the slope of the ΔRQ vs. ΔNEFA relationship was steeper in High_IS vs. Low_IS subjects (-0.139 ± 0.03 vs. -0.025 ± 0.03 RQ·mmol·l, respectively; P < 0.05), with similar intercepts. We conclude that in subjects with High_IS lipid-to-carbohydrate oxidation ratio adapts to the increased circulating NEFA availability during exercise. Such MetFlex-lip appears impaired in subjects with Low_IS. Whether a cause-effect relationship exists between impaired MetFlex-lip and low insulin sensitivity remains to be determined.

Keyword: oxygen

Geraniol improves endothelial function by inhibiting NOX-2 derived oxidative stress in high fat diet fed mice.

Endothelial dysfunction occurs in obese patients and high-fat diet (HFD) fed experimental animals. While geraniol has been reported to ameliorate inflammation and oxidative stress, inhibit tumor cell proliferation, and improve atherosclerosis, its direct effect on endothelial function remains uncharacterized. The present study therefore investigated the effect of geraniol on endothelial function in HFD mice and its underlying mechanisms. C57 BL/6 mice were fed an HFD (n\xa0=\xa040) or a normal diet (n\xa0=\xa020) for 8 weeks. HFD fed mice then were randomized to intraperitoneal treatment with geraniol (n\xa0=\xa020) or vehicle (n\xa0=\xa020) for another 6 weeks. Acetylcholine (Ach)-induced endothelial dependent vasorelaxation was measured on wire myography; reactive species (ROS) generation was assessed by fluorescence imaging, and NADPH oxidases (NOXs) and adhesive molecules VCAM-1 and ICAM-1 protein expression by western blotting. Geraniol improved endothelial function in HFD fed mice, as evidenced by its: 1. restoring endothelial dependent vasorelaxation induced by Ach, and reversing increased VCAM-1 and ICAM-1 expression; 2. attenuating HFD induced increased serum TBARS and aortic ROS generation; and 3. downregulating aortic NOX-2 expression in both HFD fed mice and in treated endothelial cells. Geraniol therefore protects against endothelial dysfunction induced by HFD through reducing NOX-2 associated ROS generation.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: oxygen

PEA and luteolin synergistically reduce mast cell-mediated toxicity and elicit neuroprotection in cell-based models of brain ischemia.

The combination of palmitoylethanolamide (PEA), an endogenous fatty amide belonging to the family of the N-acylethanolamines, and the flavonoid luteolin has been found to exert neuroprotective activities in a variety of mouse models of neurological disorders, including brain ischemia. Indirect findings suggest that the two molecules can reduce the activation of mastocytes in brain ischemia, thus modulating crucial cells that trigger the inflammatory cascade. Though, no evidence exists about a direct effect of PEA and luteolin on mast cells in experimental models of brain ischemia, either used separately or in combination. In order to fill this gap, we developed a novel cell-based model of severe brain ischemia consisting of primary mouse cortical neurons and cloned mast cells derived from mouse fetal liver (MC/9 cells) subjected to and glucose deprivation (OGD). OGD exposure promoted both mast cell degranulation and the release of lactate dehydrogenase (LDH) in a time-dependent fashion. MC/9 cells exacerbated neuronal damage in neuron-mast cells co-cultures exposed to OGD. Likewise, the conditioned medium derived from OGD-exposed MC/9 cells induced significant neurotoxicity in control primary neurons. PEA and luteolin pre-treatment synergistically prevented the OGD-induced degranulation of mast cells and reduced the neurotoxic potential of MC/9 cells conditioned medium. Finally, the association of the two drugs promoted a direct synergistic neuroprotection even in pure cortical neurons exposed to OGD. In summary, our results indicate that mast cells release neurotoxic factors upon OGD-induced activation. The association PEA-luteolin actively reduces mast cell-mediated neurotoxicity as well as pure neurons susceptibility to OGD.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: oxygen

Orexin A attenuates -induced hypothalamic cell death.

(PA), an abundant dietary saturated fatty , contributes to obesity and hypothalamic dysregulation in part through increase in oxidative stress, insulin resistance, and neuroinflammation. Increased production of reactive species (ROS) as a result of PA exposure contributes to the onset of neuronal apoptosis. Additionally, high fat diets lead to changes in hypothalamic gene expression profiles including suppression of the anti-apoptotic protein B cell lymphoma 2 (Bcl-2) and upregulation of the pro-apoptotic protein B cell lymphoma 2 associated X protein (Bax). Orexin A (OXA), a hypothalamic peptide important in obesity resistance, also contributes to neuroprotection. Prior studies have demonstrated that OXA attenuates oxidative stress induced cell death. We hypothesized that OXA would be neuroprotective against PA induced cell death. To test this, we treated an immortalized hypothalamic cell line (designated mHypoA-1/2) with OXA and PA. We demonstrate that OXA attenuates PA-induced hypothalamic cell death via reduced caspase-3/7 apoptosis, stabilization of Bcl-2 gene expression, and reduced Bax/Bcl-2 gene expression ratio. We also found that OXA inhibits ROS production after PA exposure. Finally, we show that PA exposure in mHypoA-1/2 cells significantly reduces basal respiration, maximum respiration, ATP production, and reserve capacity. However, OXA treatment reverses PA-induced changes in intracellular metabolism, increasing basal respiration, maximum respiration, ATP production, and reserve capacity. Collectively, these results support that OXA protects against PA-induced hypothalamic dysregulation, and may represent one mechanism through which OXA can ameliorate effects of obesogenic diet on brain health.Published by Elsevier Inc.

Keyword: oxygen

Pterostilbene reverses mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 diabetes and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: oxygen

Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles.

We developed a method that allows for real-time assessment of cellular metabolism in isolated, intact long skeletal muscle fibre bundles from adult mice. This method can be used to study changes in mitochondrial function and fuel utilisation in live skeletal muscle fibre bundles. Our method enables flexibility in experimental design and high-throughput assessment of mitochondrial parameters in isolated skeletal muscle fibre bundles. Extensor digitorum longus (EDL) fibre bundles obtained from chronic high-fat diet fed mice had lower basal consumption under FCCP-induced maximal respiration, when compared to control chow-fed mice. EDL fibre bundles obtained from chronic high-fat diet fed mice had enhanced mitochondrial oxidation capacity under FCCP-induced maximal respiration, when compared to control chow-fed mice.Metabolic dysfunction in skeletal muscle contributes to the aetiology and development of muscle diseases and metabolic diseases. As such, assessment of skeletal muscle cellular bioenergetics provides a powerful means to understand the role of skeletal muscle metabolism in disease and to identify possible therapeutic targets. Here, we developed a method that allows for the real-time assessment of cellular respiration in intact skeletal muscle fibre bundles obtained from the extensor digitorum longus (EDL) muscle of adult mice. Using this method, we assessed the contribution of ATP turnover and proton leak to basal mitochondrial consumption rate (OCR). Our data demonstrate that the mitochondria in EDL fibres are loosely coupled. Moreover, in the presence of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), we show that palmitate exposure induced comparable peak OCR and higher total OCR in EDL fibre bundles when compared to pyruvate exposure, suggesting that fatty acids might be a more sustainable fuel source for skeletal muscle when mitochondria are driven to maximal respiration. Application of this method to EDL fibre bundles obtained from chronic high-fat diet fed mice revealed lower basal OCR and enhanced mitochondrial oxidation capacity in the presence of FCCP when compared to the chow-diet fed control mice. By using a 96-well microplate format, our method provides a flexible and efficient platform to investigate mitochondrial parameters of intact skeletal muscle fibres obtained from adult mice.© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

Keyword: oxygen

Eicosapentaenoic Potentiates Brown Thermogenesis through FFAR4-dependent Up-regulation of miR-30b and miR-378.

Emerging evidence suggests that n-3 polyunsaturated fatty acids (PUFA) promote brown adipose tissue thermogenesis. However, the underlying mechanisms remain elusive. Here, we hypothesize that n-3 PUFA promotes brown adipogenesis by modulating miRNAs. To test this hypothesis, murine brown preadipocytes were induced to differentiate the fatty acids of , oleate, or eicosapentaenoic (EPA). The increases of brown-specific signature genes and consumption rate by EPA were concurrent with up-regulation of miR-30b and 378 but not by oleate or . Next, we hypothesize that free fatty receptor 4 (Ffar4), a functional receptor for n-3 PUFA, modulates miR-30b and 378. Treatment of Ffar4 agonist (GW9508) recapitulated the thermogenic activation of EPA by increasing consumption rate, brown-specific marker genes, and miR-30b and 378, which were abrogated in Ffar4-silenced cells. Intriguingly, addition of the miR-30b mimic was unable to restore EPA-induced Ucp1 expression in Ffar4-depleted cells, implicating that Ffar4 signaling activity is required for up-regulating the brown adipogenic program. Moreover, blockage of miR-30b or 378 by locked nucleic inhibitors significantly attenuated Ffar4 as well as brown-specific signature gene expression, suggesting the signaling interplay between Ffar4 and miR-30b/378. The association between miR-30b/378 and brown thermogenesis was also confirmed in fish oil-fed C57/BL6 mice. Interestingly, the Ffar4 agonism-mediated signaling axis of Ffar4-miR-30b/378-Ucp1 was linked with an elevation of cAMP in brown adipocytes, similar to cold-exposed or fish oil-fed brown fat. Taken together, our work identifies a novel function of Ffar4 in modulating brown adipogenesis partly through a mechanism involving cAMP activation and up-regulation of miR-30b and miR-378.© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: oxygen

Disturbance of mitochondrial functions provoked by the major long-chain 3-hydroxylated fatty acids accumulating in MTP and LCHAD deficiencies in skeletal muscle.

The pathogenesis of the muscular symptoms and recurrent rhabdomyolysis that are commonly manifested in patients with mitochondrial trifunctional protein (MTP) and long-chain 3-hydroxy-acyl-CoA dehydrogenase (LCHAD) deficiencies is still unknown. In this study we investigated the effects of the major long-chain monocarboxylic 3-hydroxylated fatty acids (LCHFA) accumulating in these disorders, namely 3-hydroxytetradecanoic (3HTA) and 3-hydroxypalmitic (3HPA) acids, on important mitochondrial functions in rat skeletal muscle mitochondria. 3HTA and 3HPA markedly increased resting (state 4) and decreased ADP-stimulated (state 3) and CCCP-stimulated (uncoupled) respiration. 3HPA provoked similar effects in permeabilized skeletal muscle fibers, validating the results obtained in purified mitochondria. Furthermore, 3HTA and 3HPA markedly diminished mitochondrial membrane potential, NAD(P)H content and Ca(2+) retention capacity in Ca(2+)-loaded mitochondria. Mitochondrial permeability transition (mPT) induction probably underlie these effects since they were totally prevented by cyclosporin A and ADP. In contrast, the dicarboxylic analogue of 3HTA did not alter the tested parameters. Our data strongly indicate that 3HTA and 3HPA behave as metabolic inhibitors, uncouplers of oxidative phosphorylation and mPT inducers in skeletal muscle. It is proposed that these pathomechanisms disrupting mitochondrial homeostasis may be involved in the muscle alterations characteristic of MTP and LCHAD deficiencies.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Argirein alleviates vascular endothelial insulin resistance through suppressing the activation of Nox4-dependent O production in diabetic rats.

Insulin resistance in endothelial cells contributes to the development of cardiovascular disease in type 2 diabetes mellitus (T2DM). Therefore, there are great potential clinical implications in developing pharmacological interventions targeting endothelial insulin resistance. Our previous studies indicated that argirein which was developed by combining rhein with L-arginine by a hydrogen bond, could substantially relieved stress related exacerbation of cardiac failure and alleviated cardiac dysfunction in T2DM, which was associated with suppressing NADPH oxidase activity. However, it is unclear whether argirein treatment attenuates the vascular lesion and dysfunction in T2DM and its underlying mechanisms.The rat aortic endothelial cells (RAECs) were used to treat with (PA), a most common saturated free fatty , which could induce insulin resistance. It was showed that argirein increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of IRS-1-ser307 and AKT-ser473, consequently resulting in the increase of the production of eNOS and NO in PA-induced RAECs. We further found that argirein blocked the Nox4-dependent superoxide (O) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vitro, argirein increased the release of endothelial NO to relieve the vasodilatory response to acetylcholine and insulin, and restored the expression of Nox4 and pIRS-1-ser307 in the aorta endothelium of high-fat diet (HFD)-fed rats following an injection of streptozocin (STZ).These results suggested that argirein could improve endothelial insulin resistance which was attributed to inhibiting Nox4-dependent redox signaling in RAECs. These studies thus revealed the novel effect of argirein to prevent the vascular complication in T2DM.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: oxygen

RNASET2 is required for ROS propagation during oxidative stress-mediated cell death.

RNASET2 is a ubiquitously expressed acidic ribonuclease that has been implicated in diverse pathophysiological processes including tumorigeneis, vitiligo, asthenozoospermia, and neurodegeneration. Prior studies indicate that RNASET2 is induced in response to oxidative stress and that overexpression of RNASET2 sensitizes cells to reactive species (ROS)-induced cell death through a mechanism that is independent of catalytic activity. Herein, we report a loss-of-function genetic screen that identified RNASET2 as an essential gene for lipotoxic cell death. Haploinsufficiency of RNASET2 confers increased antioxidant capacity and generalized resistance to oxidative stress-mediated cell death in cultured cells. This function is critically dependent on catalytic activity. Furthermore, knockdown of RNASET2 in the Drosophila fat body confers increased survival in the setting of oxidative stress inducers. Together, these findings demonstrate that RNASET2 regulates antioxidant tone and is required for physiological ROS responses.

Keyword: oxygen

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak consumption (Peak VO2), uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: oxygen

Role of SGK1 for fatty uptake, cell survival and radioresistance of NCI-H460 lung cancer cells exposed to acute or chronic cycling severe hypoxia.

Unsaturated fatty acids (FA) are required for cancer cell growth. In normoxia cells can generate unsaturated FA from saturated stearic and by desaturation. However, since the desaturation step is -dependent hypoxic cancer cells display an increased dependence on the uptake of unsaturated FA. Up to now the mechanism of increased FA uptake in hypoxia is largely unknown. Here we aimed to study the role of human serum and glucocorticoid-inducible kinase (SGK1) in the regulation of FA uptake in cancer cells exposed to acute or chronic cycling hypoxia and explore its use as target for the radiosensitization of hypoxic cancer cells.The effect of SGK1-inhibition (GSK650394) on NCI-H460 lung adenocarcinoma cells exposed to normoxia, acute or chronic cycling hypoxia was analyzed under standard and serum-deprived conditions by short-term proliferation, apoptosis and cell death assays. The impact of SGK1-inhibition on radiation sensitivity was determined by standard colony formation assays. The effect of GSK650394 on FA uptake was quantified by measuring intracellular accumulation of fluorescent FA (C1-BODIPY®-C12).Exposure to acute or chronic cycling hypoxia was associated with up-regulated expression of SGK1 in NCI-H460 cells, increased uptake of FA from the culture medium, and increased sensitivity to serum deprivation. Survival of serum-deprived hypoxic NCI-H460 cells was rescued by the addition of the unsaturated FA, oleic , whereas the saturated FA, was highly toxic to the hypoxic cancer cells. Interestingly, SGK1 inhibition abrogated the rescue effect of oleic in serum-deprived hypoxic cancer cells and this effect was associated with a reduction in FA uptake particularly in anoxia-tolerant cancer cells exposed to severe hypoxia. Finally, SKG1 inhibition decreased long-term survival and potently sensitized the parental and anoxia-tolerant NCI-H460 cells to the cytotoxic effects of ionizing radiation in normoxia as well as the anoxia-tolerant cancer cells in severe hypoxia.Our data suggest that SGK1 plays a role in the regulation of FA uptake that becomes essential under conditions of acute or chronic cycling hypoxia. We assume that SGK1 may represent a promising therapeutic target for the eradication of hypoxic cancer cells.

Keyword: oxygen

Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB.

Endothelial oxidative injury is a key event in the pathogenesis of atherosclerosis (AS). Resveratrol (RSV) attenuates the oxidative injury in human umbilical vein endothelial cells (HUVECs). Autophagy is critical for the RSV-induced protective effects. However, the exact underlying mechanisms haven\'t been completely elucidated. Thus, we aimed to explore the role of autophagy of the anti-oxidation of RSV and the underlying mechanism in (PA)-stimulated HUVECs.HUVECs were pretreated with 10\u2009μM of RSV for 2\u2009h and treated with 200\u2009μM of PA for an additional 24\u2009h. Cell viability, intracellular reactive species (ROS) and malondialdehyde (MDA) levels were estimated with a microplate reader and confocal microscope. Autophagosomes were analyzed by transmission electron microscopy, while lysosomes by confocal microscopy. The expression of transcription factor EB (TFEB) and related genes were quantified by qRT-PCR assay. Furthermore, TFEB levels, autophagy, and lysosomes were examined by western blot assay.RSV pretreatment suppressed the PA-induced decline in cell viability and elevation in ROS and MDA levels in HUVECs. RSV pretreatment also increased LC3 production and P62 degradation while promoted the autophagosomes formation. However, 3-methyladenine (3-MA) treatment attenuated RSV-induced autophagy. RSV pretreatment upregulated the TFEB and TFEB-modulated downstream genes expression in a concentration-dependent manner. Additionally, in cells transfected with TFEB small interfering RNA, RSV-induced TFEB expression and subsequent autophagy were abolished. Meanwhile, the TFEB-modulated genes expression, the lysosomes formation and the RSV-induced anti-oxidation were suppressed.In HUVECs, RSV attenuates endothelial oxidative injury by inducing autophagy in a TFEB-dependent manner.

Keyword: oxygen

Type 2 Diabetes Promotes Cell Centrosome Amplification via AKT-ROS-Dependent Signalling of ROCK1 and 14-3-3σ.

Type 2 diabetes is associated with oxidative stress and DNA damage which can cause centrosome amplification. Thus, the study investigated centrosome amplification in type 2 diabetes and the underlying mechanisms.Centrosome numbers in human peripheral blood mononuclear blood cells (PBMC) from healthy subjects and patients with type 2 diabetes were compared to access the association between type 2 diabetes and centrosome amplification. Colon cancer cells were used to investigate the molecular mechanisms underlying the centrosome amplification triggered by high glucose, insulin and . Western blot analysis was used to quantify the level of protein and protein phosphorylation. Immunofluorescent staining was performed to detect centrosomes. ROS was quantified using flow cytometry technique. Transcriptpmic profiling was performed using Illumina HiSeqTM500 platform.We found that centrosome amplification was increased PBMC from the type 2 diabetic patients, which correlated with the levels of fasting blood glucose and HbA1c. High glucose, insulin and , alone or in combinations, induced ROS production and centrosome amplification. Together, they increased AKT activation as well as the expression, binding and centrosome translation of ROCK1 and 14-3-3σ. Results from further analyses showed that AKT-ROS-dependent upregulations of expression, binding and centrosome translocation of ROCK1 and 14-3-3σ was the molecular pathway underlying the centrosome amplification in vitro triggered by high glucose, insulin and . Moreover, the key in vitro molecular signalling events activated by high glucose, insulin and were verified in PBMC from the patients with type 2 diabetes.Our results show that type 2 diabetes promotes cell centrosome amplification, and suggest that the diabetic pathophysiological factors-activated AKT-ROS-dependent signalling of ROCK1 and 14-3-3σ is the underlying molecular mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: oxygen

Downregulation of sirtuin 3 by increases the oxidative stress, impairment of mitochondrial function, and apoptosis in liver cells.

Elevated levels of saturated fatty acids show a strong cytotoxic effect in liver cells. Sirtuin 3 (SIRT3), a mitochondrially localized member of NAD -dependent deacetylase has been shown to protect hepatocytes against the oxidative stress. The role of SIRT3 on the cytotoxicity caused by fatty acids in liver cells is not fully understood. The aim of this study was to evaluate the expression level of SIRT3, oxidative stress, and mitochondrial impairments in human hepatoma HepG2 cells exposed to (PA). Our results showed that PA treatment caused the deposition of lipid droplets and resulted in an increased expression of tumor necrosis factor-α in a dose-dependent manner. Excessive accumulation of PA induces the reactive species formation and apoptosis while dissipating the mitochondrial transmembrane potential. The level of SIRT3 expression in both nuclear and mitochondrial fractions in HepG2 cells was decreased with the increase in PA concentrations. However, in the cytosolic fraction, the SIRT3 was undetectable. In conclusion, our results showed that PA caused an increase in inflammation and oxidative stress in HepG2 cells. The exposure of PA also resulted in the decline in transmembrane potential and an increase in apoptosis. The underexpression of nuclear and mitochondrial SIRT3 by PA suggests that the PA target the process that regulates the stress-related gene expression and mitochondrial functions.© 2019 Wiley Periodicals, Inc.

Keyword: oxygen

Rare ginsenosides ameliorate lipid overload-induced myocardial insulin resistance via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 diabetes. However, their pharmacological studies regarding myocardial fuel metabolism and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel metabolism and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high fat diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive species (ROS) production were measured by fluorescence labeling. Other assessments including consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of lipid overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: oxygen

Metabolic Alterations and the Protective Effect of Punicalagin Against Glutamate-Induced Oxidative Toxicity in HT22 Cells.

Oxidative stress is involved in many neurological diseases, including Alzheimer\'s disease. Punicalagin (PC) is a hydrolysable polyphenol derived from Punica granatum and a potent antioxidant. In this study, the neuroprotective effect of PC on glutamate-induced oxidative stress was evaluated in the mouse hippocampal cell line, HT22. PC treatment protected HT22 cells from glutamate-induced cell death in a concentration-dependent manner, potentially attenuated glutamate-induced intracellular reactive species (ROS) and restored the mitochondrial membrane depolarization. Metabolic alterations after glutamate-induced oxidative stress and the protective effect of PC were evaluated with HPLC and GC-MS profiling methods with multivariate statistical analyses. Alterations in ten metabolites were identified, including amino acids, aspartic , asparagine, threonine, anserine, cysteine, tryptophan, lysine, as well as fatty acids , stearic , and palmitoleic . Metabolic pathway analysis revealed the involvement of multiple affected pathways, such as cysteine and methionine metabolism, tryptophan metabolism, alanine, aspartate, and glutamate and fatty oxidation. These results clearly demonstrate that PC is a promising therapeutic agent for oxidative stress-associated diseases.

Keyword: oxygen

-induced autophagy increases reactive species via the Ca/PKCα/NOX4 pathway and impairs endothelial function in human umbilical vein endothelial cells.

It is well known that the lipotoxic mechanism of (PA), a main constituent of triglyceride, is dependent on reactive species (ROS). Recently, it has also been reported that PA is an autophagy inducer. However, the causal association and underlying mechanism of induced autophagy and ROS in PA toxicity remain unclear. The present study demonstrates for the first time that PA-induced autophagy enhances ROS generation via activating the calcium ion/protein kinase Cα/nicotinamide adenine dinucleotide phosphate oxidase 4 (Ca/PKCα/NOX4) pathway in human umbilical vein endothelial cells (HUVECs). It was revealed that PA treatment resulted in a significant increase in ROS generation and autophagic activity, leading to endothelial dysfunction as indicated by downregulated nitric oxide synthesis, decreased capillary-like structure formation and damaged cell repair capability. Furthermore, PA effectively activated the Ca/PKCα/NOX4 pathway, which is indicative of upregulated cytosolic Ca levels, activated PKCα and increased NOX4 protein expression. 3-Methyladenine was then used to inhibit autophagy, which significantly reduced PA-induced ROS generation and blocked the Ca/PKCα/NOX4 pathway. The endothelial dysfunction caused by PA was ameliorated by downregulating ROS generation using a NOX4 inhibitor. In conclusion, PA-induced autophagy contributes to endothelial dysfunction by increasing oxidative stress via the Ca/PKCα/NOX4 pathway in HUVECs.

Keyword: oxygen

Co-overexpression of native phospholipid-biosynthetic genes plsX and plsC enhances lipid production in Synechocystis sp. PCC 6803.

The overexpression of native plsX and plsC genes involving in fatty /phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and evolution rate of all overexpressing (OX) strains were normally similar as wild type. For fatty compositions, saturated fatty , in particular (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated fatty acids were observed, specifically linoleic (18:2) and alpha-linolenic (18:3). The plsC/plsX-overexpressing (OX\u2009+\u2009XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX\u2009+\u2009XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.

Keyword: oxygen

miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: oxygen

Biofuel and co-products from algae solvent extraction.

This study reports a novel method of using algae biomass as a source of lipid and various other co-products. Solvent-based extraction techniques could yield a number of products simultaneously. Further, the study focuses on all possible characterization and utilization of the three layers obtained from chloroform-methanol extraction of lipids. The lipid from the chloroform layer was transesterified for Fatty methyl esters (FAME) production. The fatty methyl esters derived by oleic , , linoleic and phytol were majorly analysed by GC-MS. The methanol layer was analysed with HPLC and stachyose, maltotriose, glucose, fructose, acetic , butyric , DMSO, glycerol were identified. The cell debris was further physically activated, and physiochemical properties of raw algae, residual algae and algae bio-char were compared. Spectrum peaks of FTIR study identified many alkyl‒halide stretches. Similarly, EDX analysed the presence of carbon, nitrogen, , potassium, chlorine, calcium, iron, magnesium and phosphorus. The SEM reveals that residual algae was comparatively crystalline and hence could not be utilized directly as an adsorbent. Therefore, further physical treatment was applied, and methylene blue dye adsorption study was also conducted to know the time and capacity of biochar as an adsorbent. However, organic and mineral enriched biomass could be used directly as fertilizer for agricultural purposes.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: oxygen

BNIP3 induction by hypoxia stimulates FASN-dependent free fatty production enhancing therapeutic potential of umbilical cord blood-derived human mesenchymal stem cells.

Mitophagy under hypoxia is an important factor for maintaining and regulating stem cell functions. We previously demonstrated that fatty synthase (FASN) induced by hypoxia is a critical lipid metabolic factor determining the therapeutic efficacy of umbilical cord blood-derived human mesenchymal stem cells (UCB-hMSCs). Therefore, we investigated the mechanism of a major mitophagy regulator controlling lipid metabolism and therapeutic potential of UCB-hMSCs. This study revealed that Bcl2/adenovirus E1B 19kDa protein-interacting protein 3 (BNIP3)-dependent mitophagy is important for reducing mitochondrial reactive species accumulation, anti-apoptosis, and migration under hypoxia. And, BNIP3 expression was regulated by CREB binding protein-mediated transcriptional actions of HIF-1α and FOXO3. Silencing of BNIP3 suppressed free fatty (FFA) synthesis regulated by SREBP1/FASN pathway, which is involved in UCB-hMSC apoptosis via caspases cleavage and migration via cofilin-1-mediated F-actin reorganization in hypoxia. Moreover, reduced mouse skin wound-healing capacity of UCB-hMSC with hypoxia pretreatment by BNIP3 silencing was recovered by . Collectively, our findings suggest that BNIP3-mediated mitophagy under hypoxia leads to FASN-induced FFA synthesis, which is critical for therapeutic potential of UCB-hMSCs with hypoxia pretreatment.Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: oxygen

TLR4 knockout attenuated high fat diet-induced cardiac dysfunction via NF-κB/JNK-dependent activation of autophagy.

Obesity is commonly associated with a low grade systemic inflammation, which may contribute to the onset and development of myocardial remodeling and contractile dysfunction. Toll-like receptor 4 (TLR4) plays an important role in innate immunity and inflammation although its role in high fat diet-induced obesity cardiac dysfunction remains elusive. This study was designed to examine the effect of TLR4 ablation on high fat diet intake-induced cardiac anomalies, if any, and underlying mechanism(s) involved. Wild-type (WT) and TLR4 knockout mice were fed normal or high fat (60% calorie from fat) diet for 12weeks prior to assessment of mechanical and intracellular Ca properties. The inflammatory signaling proteins (TLR4, NF-κB, and JNK) and autophagic markers (Atg5, Atg12, LC3B and p62) were evaluated. Our results revealed that high fat diet intake promoted obesity, marked decrease in fractional shortening, and cardiomyocyte contractile capacity with dampened intracellular Ca release and clearance, elevated ROS generation and oxidative stress as measured by aconitase activity, the effects of which were significantly attenuated by TLR4 knockout. In addition, high fat intake downregulated levels of Atg5, Atg12 and LC3B, while increasing p62 accumulation. TLR4 knockout itself did not affect Atg5, Atg12, LC3B and p62 levels while it reconciled high fat diet intake-induced changes in autophagy. In addition, TLR4 knockout alleviated high fat diet-induced phosphorylation of IKKβ, JNK and mTOR. In vitro study revealed that suppressed cardiomyocyte contractile function, the effect of which was inhibited the TLR4 inhibitor CLI-095, the JNK inhibitor AS601245 or the NF-κB inhibitor Celastrol. Taken together, these data showed that TLR4 knockout ameliorated high fat diet-induced cardiac contractile and intracellular Ca anomalies through inhibition of inflammation and ROS, possibly through a NF-κB/JNK-dependent activation of autophagy. This article is part of a Special Issue entitled: Genetic and epigenetic control of heart failure - edited by Jun Ren & Megan Yingmei Zhang.Copyright © 2017. Published by Elsevier B.V.

Keyword: oxygen

Ameliorative effect of panaxynol on the reduction in high-molecular-weight adiponectin secretion from 3T3-L1 adipocytes treated with acids.

Reduced plasma levels of the high-molecular weight (HMW) form of adiponectin, rather than total adiponectin levels, have been shown to be closely associated with various metabolic diseases including insulin resistance, type 2 diabetes, and cardiovascular disease. Therefore, we sought to explore active, naturally occurring compounds that promote the recovery of HMW adiponectin secretion suppressed by in our model. A total of 90 crude drug extracts were screened for the ability to augment HMW adiponectin secretion from 3T3-L1 adipocytes treated with . Panaxynol was isolated from Saposhnikovia divaricata as an active compound with HMW adiponectin promoting properties. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists are reported to increase the secretion of HMW adiponectin, although the effects of panaxynol were found to be independent of PPARγ activation. When the underlying mechanisms were further examined, panaxynol was found to inhibit the -induced downregulation of forkhead box O1 (FoxO1) protein, and the anti-lipotoxic effects were abolished by a FoxO1 inhibitor. Furthermore, CCAAT/enhancer-binding protein-α (C/EBPα) mRNA levels were also increased by panaxynol. Reactive species have critical roles in the reduction in HMW adiponection secretion by ; however, panaxynol reduced this increase in reactive species generation, followed by reductions in markers of endoplasmic reticulum stress and inflammation. Taken together, these findings suggest that panaxynol ameliorates the impaired HMW adiponection secretion in adipocytes treated with by restoring FoxO1 expression, owing to inhibition of reactive species generation, in a PPARγ-independent manner.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: oxygen

ShenFu Preparation Protects AML12 Cells Against -Induced Injury Through Inhibition of Both JNK/Nox4 and JNK/NFκB Pathways.

Nonalcoholic steatohepatitis includes steatosis along with liver inflammation, hepatocyte injury and fibrosis. In this study, we investigated the protective role and the potential mechanisms of a traditional Chinese medicine ShenFu (SF) preparation in an in vitro hepatic steatosis model.In (PA)-induced murine hepatic AML12 cell injury, effects of SF preparation on cellular apoptosis and intracellular triglyceride (iTG) level were assessed using TUNEL and TG Colorimetric Assay. Reactive species (ROS) and mitochondrial membrane potential (MMP) levels were measured using DCF and JC-1 assay. Cytokine levels were evaluated using ELISA assay. Immunoblot was used to compare the activation level of c-Jun N terminal kinase (JNK), NADPH oxidase (Nox4), and NFκB pathways.Addition of SF preparation prevented PA-mediated increase of apoptosis and iTG as well as IL-8 and IL-6. In PA-treated cell, SF preparation reduced the level of Nox4 and ROS, while increasing the level of MMP and the expression of manganese superoxide dismutase (MnSOD) and catalase, indicating emendation of mitochondrial dysfunction. Nox4 inhibitor GKT137381 prevented PA-induced increase of ROS and apoptosis, while decreasing iTG slightly and not influencing the level of IL-8 and IL-6. SF preparation prevented PA-induced upregulation of phospho-JNK. JNK inhibitor SP600125 prevented PA-mediated increase of Nox4, IL-8, IL-6 and iTG. Nuclear translocation of NFκB/p65 was detected in PA-treated cells, which was prevented by SF preparation. An IκB degradation inhibitor, BAY11-7082, prevented PA-induced increase of IL-8 and IL-6 as well as iTG, whereas it only decreased ROS levels slightly and showed no influence on cellular apoptosis.SF preparation shows a beneficial role in prevention of hepatocyte injury by attenuating oxidative stress and cytokines production at least partially through inhibition of JNK/Nox4 and JNK/NFκB pathway, respectively.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: oxygen

Sonodynamic therapy inhibits palmitate-induced beta cell dysfunction via PINK1/Parkin-dependent mitophagy.

In type 2 diabetes mellitus (T2DM), the overload of glucose and lipids can promote oxidative stress and inflammatory responses and contribute to the failure of beta cells. However, therapies that can modulate the function of beta cells and thus prevent their failure have not been well explored. In this study, beta cell injury model was established with (PA) to simulate the lipotoxicity (high-fat diet) found in T2DM. Sonodynamic therapy (SDT), a novel physicochemical treatment, was applied to treat injured beta cells. We found that SDT had specific effects on mitochondria and induced transient large amount of mitochondrial reactive species (ROS) production in beta cells. SDT also improved the morphology and function of abnormal mitochondria, inhibited inflammatory response and reduced beta cell dysfunction. The improvement of mitochondria was mediated by PINK1/Parkin-dependent mitophagy. Additionally, SDT rescued the transcription of PINK1 mRNA which was blocked by PA treatment, thus providing abundant PINK1 for mitophagy. Moreover, SDT also increased insulin secretion from beta cells. The protective effects of SDT were abrogated when mitophagy was inhibited by cyclosporin A (CsA). In summary, SDT potently inhibits lipotoxicity-induced beta cell failure via PINK1/Parkin-dependent mitophagy, providing theoretical guidance for T2DM treatment in aspects of islet protection.

Keyword: oxygen

Polysaccharide from Rubus chingii Hu affords protection against -induced lipotoxicity in human hepatocytes.

(PA) is known to induce lipotoxicity, a metabolic syndrome as a result of lipid accumulation in multiple cell lines. Bioactive phytochemicals derived from vegetables and fruits have gained increasing attention owing to their potential on suppressing the detrimental effect of excessive PA accumulation. However, the protective effect of natural phytochemicals derived from Rubus chingii Hu, a kind of fruit widely grown in China, against PA-induced lipotoxicity is still uncleared. In the present study, we therefore extracted the polysaccharide from Rubus chingii Hu, and identified its chemical structure. Structural characterization by HPLC, HPGPC, IR spectroscopy and GC indicated that the polysaccharide mainly consists of galacturonic and arabinose with copious 1\u202f→\u202f2 glycosidic linkages in its backbone. In addition, our results showed the cytoprotective effect of the polysaccharide against PA-induced lipotoxicity in normal human hepatocyte cell line L02. Further study indicated that the polysaccharide mitigated oxidative stress through impeding cellular reactive species (ROS) accumulation, alleviating mitochondrial membrane potential (MMP) collapse and attenuating glutathione (GSH) reduction. Overall, this study revealed that Rubus chingii Hu polysaccharide was capable of effectively alleviating -induced lipotoxicity, which provided a novel perspective of the health-promoting potential of isolated polysaccharide.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: oxygen

induces neurotoxicity and gliatoxicity in SH-SY5Y human neuroblastoma and T98G human glioblastoma cells.

Obesity-related central nervous system (CNS) pathologies like neuroinflammation and reactive gliosis are associated with high-fat diet (HFD) related elevation of saturated fatty acids like (PA) in neurons and astrocytes of the brain.Human neuroblastoma cells SH-SY5Y (as a neuronal model) and human glioblastoma cells T98G (as an astrocytic model), were treated with 100-500 µM PA, oleic (OA) or lauric (LA) for 24 h or 48 h, and their cell viability was assessed by 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of stable overexpression of γ-synuclein (γ-syn), a neuronal protein recently recognized as a novel regulator of lipid handling in adipocytes, and transient overexpression of Parkinson\'s disease (PD) α-synuclein [α-syn; wild-type (wt) and its pathogenic mutants A53T, A30P and E46K] in SH-SY5Y and T98G cells, were also evaluated. The effects of co-treatment of PA with paraquat (PQ), a Parkinsonian pesticide, and leptin, a hormone involved in the brain-adipose axis, were also assessed. Cell death mode and cell cycle were analyzed by Annexin V/PI flow cytometry. Reactive species (ROS) level was determined using 2\',7\'-dichlorofluorescien diacetate (DCFH-DA) assay and lipid peroxidation level was determined using thiobarbituric reactive substances (TBARS) assay.MTT assay revealed dose- and time-dependent PA cytotoxicity on SH-SY5Y and T98G cells, but not OA and LA. The cytotoxicity was significantly lower in SH-SY5Y-γ-syn cells, while transient overexpression of wt α-syn or its PD mutants (A30P and E46K, but not A53T) modestly (but still significantly) rescued the cytotoxicity of PA in SH-SY5Y and T98G cells. Co-treatment of increasing concentrations of PQ exacerbated PA\'s neurotoxicity. Pre-treatment of leptin, an anti-apoptotic adipokine, did not successfully rescue SH-SY5Y cells from PA-induced cytotoxicity-suggesting a mechanism of PA-induced leptin resistance. Annexin V/PI flow cytometry analysis revealed PA-induced increase in percentages of cells in annexin V-positive/PI-negative quadrant (early apoptosis) and subG-G fraction, accompanied by a decrease in G-M phase cells. The PA-induced ROS production and lipid peroxidation was at greater extent in T98G as compared to that in SH-SY5Y.In conclusion, PA induces apoptosis by increasing oxidative stress in neurons and astrocytes. Taken together, the results suggest that HFD may cause neuronal and astrocytic damage, which indirectly proposes that CNS pathologies involving neuroinflammation and reactive gliosis could be prevented via the diet regimen.

Keyword: oxygen

Protocatechuic Ameliorated -Induced Oxidative Damage in Endothelial Cells through Activating Endogenous Antioxidant Enzymes via an Adenosine-Monophosphate-Activated-Protein-Kinase-Dependent Pathway.

Protocatechuic (PCA, 3,4-dihydroxybenzoic ), the main metabolite of anthocyanins, is widely distributed in fruits and vegetables and has been reported to possess a strong antioxidant activity. Herein, we aimed to investigate the protective effect of PCA against high (PA)-induced oxidative damage and the underling molecular mechanisms in human umbilical vein endothelial cells (HUVECs). PCA reduced the levels of intracellular reactive species and malondialdehyde and increased the activities of endogenous antioxidant enzymes, including superoxide dismutase, glutathione peroxidase 1, and heme oxygenase 1 (HO-1). Metabolomic analysis showed that PCA affected numerous metabolites, especially some of which were related with energy metabolism. PCA also upregulated the phosphorylation of adenosine-monophosphate-activated protein kinase (AMPK) at Thr through activating liver kinase B1 and then promoted the expression of p-Nrf2 and HO-1. Moreover, PCA reversed the decreased expression of peroxisome proliferator-activated receptor γ coactivator 1α and significantly increased the mitochondrial density. Collectively, these results demonstrated that PCA attenuated PA-induced oxidative damage in HUVECs via an AMPK-dependent pathway.

Keyword: oxygen

[Influence of Fatty Acids on Consumption in Isolated Cardiomyocytes of Rats with Ischemic or Diabetic Heart Disease].

one of the reasons of violation of the functional viability of the myocardium is considered to be the deprivation and lack of energy. The reason is the inhibitory effect of fatty acids on glucose oxidation. Recently, however, new data have been published proving the need for fatty acids and their importance in the maintenance and regulation of the functional activity of the myocardium in chronic pathology.to investigate the influence of free polyunsaturated and saturated fatty acids (FA) on the uptake of isolated cardiomyocytes in intact rats and animals with ischemic or diabetic heart disease.the executed non-randomized controlled study. It includied 3 groups of male rats of Wistar line (weight 250-300g) with 10 animals in each group. Myocardial infarction ("heart attack" group) was caused by ligation of the left coronary artery, diabetes ("diabetes" group)--by intraperitoneal injection of streptozotocin, and "control" group (intact animals). Myocardial infarction caused by ligation of the left coronary artery, and diabetes by intraperitoneal injection of streptozotocin. Isolated cardiac myocytes were obtained by the enzymatic method. consumption was assessed polarographically at different saturation incubation medium with ([O₂] ≤ 8 mg/l and ([O₂] ≥ 16 mg/l). Arachidonic and acids were applied as fatty acids.It is established that the introduction of the incubation medium 20 µm arachidonic or fatty significantly increased the consumption of intact cardiomyocytes of rats. Both at the ischemic and at the diabetic injury to the heart the opposite result was obtained. The most pronounced decrease in consumption was indicated in the group with diabetes mellitus.The inhibitory effect of LCD on the rate of consumption may be associated with the influence of the ischemic or diabetic injury to the heart on the barrierfunction ofmitochondrial membranes of cardiomyocytes, the activity of membrane-associated enzymes and their associated processes.

Keyword: oxygen

HAMSCs/HBMSCs coculture system ameliorates osteogenesis and angiogenesis against glucolipotoxicity.

Osteoporosis and vascular lesions induced by glucolipotoxicity are common complications of diabetes mellitus (DM). In order to deal with these complications, we designed a new therapeutic strategy, i.e. coculture system containing human amnion-derived mesenchymal stem cells (HAMSCs) and human bone marrow mesenchymal stem cells (HBMSCs). Two in\xa0vitro coculture models, transwell and mixed cocultures, were proposed for 7 days with variable HAMSCs: HBMSCs ratios. Then, supernatant from each coculture was used to reverse the deficiency of HBMSCs and human umbilical vein endothelial cells (HUVECs) impaired by high glucose and (GP). We found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation, osteogenic differentiation and superoxide dismutase (SOD) activity, as well as induced the reactive species (ROS) level in HBMSCs. Meanwhile, glucolipotoxicity suppressed cell proliferation, tube formation capacity and angiogenic potential of HUVECs. Though, HAMSCs/HBMSCs coculture system reduced HBMSCs dysfunction by antioxidant properties and promoted angiogenesis in HUVECs. The mixed HAMSCs/HBMSCs coculture at the optimal ratio of 3/1 showed significantly greater cell proliferation, antioxidant properties, osteogenic and angiogenic differentiation than HBMSCs or HUVECs alone. In conclusion, the current coculture system of HAMSCs/HBMSCs can be a potential therapeutic material for advancing bone and vascular regeneration against DM-induced glucolipotoxicity.Copyright © 2018 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: oxygen

Thermo-chemo-sonic pre-digestion of waste activated sludge for yeast cultivation to extract lipids for biodiesel production.

The low cost biosynthesis of microbial lipids are an efficient feedstock to replace plant based oil for biodiesel production. The present study objective is to explore the effect of thermo-chemo-sonic pre-digestion of municipal Waste Activated Sludge (WAS) to cultivate oleaginous L.\xa0starkeyi MTCC-1400 as a model organism to produce high yield biomass and lipid. Higher Suspended Solids (SS) reduction (20 and 15.71%) and Chemical Demand (COD) solubilization (27.6 and 22.3%) were achieved at a Specific Energy (SE) input of 5569\xa0kJ/kg for WAS digested with NaOH and KOH, respectively. The maximum biomass of 17.52\xa0g\xa0L and lipid 64.3% dwt were attained in NaOH pre-digested sample. The analyzed lipid profile exhibited high content of (45.6%) and oleic (38.7%) which are more suitable for biofuel production. Thus, these results strongly motivate the use of pre-digested WAS as an efficient and economical substrate for biodiesel production.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: oxygen

NDP-MSH reduces oxidative damage induced by in primary astrocytes.

Recent findings relate obesity to inflammation in key hypothalamic areas for body weight control. Hypothalamic inflammation has also been related to oxidative stress. (PA) is the most abundant free fatty found in food, and in vitro studies indicate that it triggers a pro-inflammatory response in the brain. Melanocortins are neuropeptides with proven anti-inflammatory and neuroprotective action mediated by melanocortin receptor 4 (MC4R), but little is known about the effect of melanocortins on oxidative stress. The aim of this study was to investigate whether melanocortins could alleviate oxidative stress induced by a high fat diet (HFD) model. We found that NDP-MSH treatment decreased PA-induced reactive species production in astrocytes, an effect blocked by the MC4R inhibitor JKC363. NDP-MSH abolished nuclear translocation of Nrf2 induced by PA and blocked the inhibitory effect of PA on superoxide dismutase (SOD) activity and glutathione levels while it also per se increased activity of SOD and γ-glutamate cysteine ligase (γ-GCL) antioxidant enzymes. However, HFD reduced hypothalamic MC4R and brain derived neurotrophic factor mRNA levels, thereby preventing the neuroprotective mechanism induced by melanocortins.© 2019 British Society for Neuroendocrinology.

Keyword: oxygen

MD2 Blockage Protects Obesity-Induced Vascular Remodeling via Activating AMPK/Nrf2.

Obesity and increased free fatty (FFA) levels are tightly linked with vascular oxidative stress and remodeling. Myeloid differentiation 2 (MD2), an important protein in innate immunity, is requisite for endotoxin lipopolysaccharide responsiveness. This study shows that (PA) also bonds to MD2, initiating cardiac inflammatory injury. However, it is not clear whether MD2 plays a role in noninflammatory systems such as obesity- and FFA-related oxidative stress involved in vascular remodeling and injury. The aim of this study is to examine whether MD2 participates in reactive species increase and vascular remodeling.Male MD2 mice and wild-type littermates with a C57BL/6 background were fed a high-fat diet (HFD) to establish obesity-induced vascular remodeling. Rat aortic endothelial cells (RAECs) and vascular smooth muscle cells (VSMCs) were treated with PA to induce oxidative stress and injury.In vivo, MD2 deficiency significantly reduced HFD-induced vascular oxidative stress, fibrosis, and remodeling, accompanied with AMP-activated kinase (AMPK) activation and nuclear factor erythroid (Nrf2) upregulation. In VSMCs and RAECs, inhibition of MD2 by neutralizing monoclonal antibody to MD2 or small interfering RNA knockdown significantly activated the AMPK/Nrf2-signaling pathway and reduced PA-induced oxidative stress and cell injury.It was demonstrated that the deletion or inhibition of MD2 protects against HFD/FFA-induced vascular oxidative stress and remodeling by activating the AMPK/Nrf2-signaling pathway.© 2017 The Obesity Society.

Keyword: oxygen

[ induces hepatocellular oxidative stress and activation of inflammasomes].

To evaluate the effect of (PA) on oxidative stress and activation of inflammasomes in hepatocytes.To test the dose-dependent effect of PA on normal murine hepatocytes AML12, the cells were treated with 0, 0.15, 0.25 and 0.4 mmol/L of (PA). The cells were also divided into blank control group, 0.25 mmol/L PA group and 0.25 mmol/L PA+N-acetylcysteine (NAC) group to examine the effect of reactive species (ROS) on the activation of inflammasomes. After 24 h of treatment, lipid accumulation, total ROS, mitochondrial ROS, expression and localization of NOX4, and expressions of inflammasomes and IL-1β were detected in the hepatocytes.Compared with the control cells, PA treatment of the cells significantly increased cytoplasmic lipid accumulation, concentrations of total ROS (12 463.09±2.72 vs 6691.23±2.45, P=0.00) and mitochondrial ROS (64.98±0.94 vs 45.04±0.92, P=0.00), and the expressions of NOX4, NLRP3, ASC, caspase-1, and IL-1β (1603.52±1.32 vs 2629.33±2.57, P=0.00). The mitochondria and NOX4 were found to be co-localized in the cytoplasm. NAC obviously reduced cellular ROS level stimulated by PA (7782.15±2.87 vs 5445.6±1.17, P=0.00) and suppressed the expressions of NLRP3, ASC and caspase-1.PA treatment can stimulate lipid accumulation in hepatocytes and induce oxidative stress through NOX4 and mitochondria pathway to activate inflammasomes and stimulate the secretion of IL-1β.

Keyword: oxygen

Tibolone Reduces Oxidative Damage and Inflammation in Microglia Stimulated with through Mechanisms Involving Estrogen Receptor Beta.

High concentrations of in plasma increase both the inflammation associated with obesity and the susceptibility to develop a neurodegenerative event. In the brain, the inflammatory response is mediated by activated microglial cells, which undergo morphological and biochemical changes and can directly affect cell viability. Recent evidence shows that the use of estrogenic compounds can control microglia-induced inflammation with promising results. In this study, we explored the actions of the synthetic steroid tibolone on BV-2 microglia cells stimulated with . Our results demonstrated that tibolone increased cell viability and reduced nuclear fragmentation and the production of reactive species, as well as preserved mitochondrial membrane potential. These effects were accompanied by reduced nuclear translocation of NF-κB p65, upregulation of neuroglobin, and improved antioxidant defense. Furthermore, estrogen receptor beta (ERβ) inhibition partially dampened tibolone\'s protective actions in BV-2 cells stimulated with . In conclusion, tibolone protects BV-2 cells by a mechanism involving ERβ and upregulation of neuroglobin.

Keyword: oxygen

Optimization of simultaneous biomass production and nutrient removal by mixotrophic Chlorella sp. using response surface methodology.

The bioprospecting of potentially mixotrophic microalgae in a constructed wetland was conducted. A locally isolated microalga, Chlorella sp., was grown to determine the effect of temperature, aeration rate, and cultivation time on simultaneous biomass production and nutrient removal from piggery wastewater using central composite design (CCD). The most important variable for the biomass productivity of Chlorella sp. was aeration rate, while that for lipid content and nutrient removal efficiency was cultivation time. Total nitrogen (TN) and total phosphorus (TP) removal efficiencies were higher than that of chemical demand (COD) from piggery wastewater. The CCD results indicate that the highest biomass productivity (79.2 mg L(-1) d(-1)) and simultaneous nutrient removal efficiency (TN 80.9%, TP 99.2%, COD 74.5%) were obtained with a cultivation temperature of 25 °C, a cultivation time of 5 days, and an air aeration rate of 1.6 L L(-1) min(-1). (C16:0) and linoleic (C18:2) were both abundant in Chlorella sp. cells under mixotrophic cultivation with piggery wastewater.

Keyword: oxygen

Human amnion-derived mesenchymal stem cells promote osteogenesis of human bone marrow mesenchymal stem cells against glucolipotoxicity.

Epidemiological evidence suggests that diabetes mellitus (DM) is an important factor in promoting periodontitis. It not only affects the attachment of connective tissue but also causes loss of alveolar bone. Hence, there is an urgent need to find an effective treatment for DM-induced bone deficiency. This study aimed to investigate the effects of human amniotic mesenchymal stem cells (HAMSCs) on the proliferation and osteogenic differentiation of DM-induced human bone marrow mesenchymal stem cells (HBMSCs). High glucose and (GP) were used to mimic DM-induced glucolipotoxicity. The proliferation levels were measured using flow cytometry. Alkaline phosphatase activity substrate assays, Alizarin red S staining, and western blotting were used to investigate osteogenic differentiation. Oxidative stress was measured by assaying the levels of reactive species. This study found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation and osteogenesis, and upregulated the oxidative stress level in HBMSCs. However, HAMSCs attenuated HBMSC dysfunction through antioxidant activity by influencing p38 mitogen-activated protein kinase and vascular endothelial growth factor secretion. In conclusion, our findings indicate that HAMSCs might be suitable for treating DM-mediated bone deficiency.

Keyword: oxygen

TERT inhibition leads to reduction of IL-6 expression induced by and interferes with the protective effects of tibolone in an astrocytic cell model.

Although it has been shown that telomerase has neuroprotective effects, mainly as a result of its non-canonical functions in neuronal cells, its role with respect to glial cells remains unknown. There is growing evidence indicating that telomerase plays an important role with respect to inflammation, especially in the regulation of pro-inflammatory cytokine gene expression. The present study aimed to evaluate the role of telomerase in an astrocyte cell model treated with (PA) and tibolone. Cell death, reactive species production and interleukin-6 expression were evaluated under telomerase inhibition with the BIBR1532 compound in T98G cells treated with tibolone and PA, using fluorometry, flow cytometry, enzyme-linked immunosorbent assays and the quantitative polymerase chain reaction. The results obtained showed that telomerase protein was increased by PA after 36\xa0hours, alone or in combination with tibolone, and that its activity was affected by PA. Telomerase inhibition reduced interleukin-6 expression and it interfered with the protective effects of tibolone on cell death. Moreover, tibolone increased Tyr707 phosphorylation in PA-treated cells. In the present study, we provide novel findings about the regulation of telomerase by PA and tibolone. Telomerase was involved in inflammation by PA and in protective effects of tibolone. Therefore, we conclude that telomerase could play a dual role in these cells.© 2019 British Society for Neuroendocrinology.

Keyword: oxygen

Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

Keyword: oxygen

NAFLD causes selective CD4(+) T lymphocyte loss and promotes hepatocarcinogenesis.

Hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death. Non-alcoholic fatty liver disease (NAFLD) affects a large proportion of the US population and is considered to be a metabolic predisposition to liver cancer. However, the role of adaptive immune responses in NAFLD-promoted HCC is largely unknown. Here we show, in mouse models and human samples, that dysregulation of lipid metabolism in NAFLD causes a selective loss of intrahepatic CD4(+) but not CD8(+) T lymphocytes, leading to accelerated hepatocarcinogenesis. We also demonstrate that CD4(+) T lymphocytes have greater mitochondrial mass than CD8(+) T lymphocytes and generate higher levels of mitochondrially derived reactive species (ROS). Disruption of mitochondrial function by linoleic , a fatty accumulated in NAFLD, causes more oxidative damage than other free fatty acids such as , and mediates selective loss of intrahepatic CD4(+) T lymphocytes. In vivo blockade of ROS reversed NAFLD-induced hepatic CD4(+) T lymphocyte decrease and delayed NAFLD-promoted HCC. Our results provide an unexpected link between lipid dysregulation and impaired anti-tumour surveillance.

Keyword: oxygen

triggers cell apoptosis in RGC-5 retinal ganglion cells through the Akt/FoxO1 signaling pathway.

Hallmarks of the pathophysiology of glaucoma are oxidative stress and apoptotic death of retinal ganglion cells (RGCs). Lipotoxicity, involving a series of pathological cellular responses after exposure to elevated levels of fatty acids, leads to oxidative stress and cell death in various cell types. The phosphatidylinositol-3-kinase/protein kinase B/Forkhead box O1 (PI3K/Akt/FoxO1) pathway is crucial for cell survival and apoptosis. More importantly, FoxO1 gene has been reported to confer relatively higher risks for eye diseases including glaucoma. However, little information is available regarding the interaction between FoxO1 and RGC apoptosis, much less a precise mechanism. In the present study, immortalized rat retinal ganglion cell line 5 (RGC-5) was used as a model to study the toxicity of (PA), as well as underlying mechanisms. We found that PA exposure significantly decreased cell viability by enhancing apoptosis in RGC-5 cells, as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. PA also induced a remarkable increase in reactive species and malondialdehyde. Moreover, PA significantly decreased the level of phospho-Akt and phospho-FoxO1 in cells. Finally, shRNA knockdown and plasmid overexpression studies displayed that downregulation of Akt protein or upregulation of FoxO1 protein augmented cell death, while knockdown of FoxO1 or overexpression of Akt1 abolished PA-induced cell death. Collectively, our results indicated that PA-induced cell death is mediated through modulation of Akt/FoxO1 pathway activity.

Keyword: oxygen

A microplate assay for measuring cell death in C2C12 cells.

The main goal of this study was to develop a straightforward and rapid microplate assay for measuring propidium iodide (PI) in C2C12 cells. The PI method has proven to be an efficient quantitative assay for analyzing cell viability through PI fluorescence analysis. Importantly, the protocol takes less than 30 min and the results are reproducible. C2C12 cells were exposed to an increasing concentration of palmitate for a period of 24 h to induce cell death, and the PI fluorescence increased in a concentration-dependent manner. Evaluation of mitochondrial function and the production of reactive species confirmed the deleterious effects of palmitate. Also, the microplate PI assay demonstrated high sensitivity, as indicated by the detection of modest fluctuations in cell viability in response to catalase overexpression in palmitate-treated cells. The microplate PI assay, therefore, offers an accurate method for use in in-vitro studies.

Keyword: oxygen

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

Inflammation markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free fatty acids (FFAs) are known to cause damage in various tissues by inducing inflammation. In this study, we investigated whether a FFA (palmitate) induces inflammation in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish liver and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including diabetes.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

A novel role for small molecule glycomimetics in the protection against lipid-induced endothelial dysfunction: Involvement of Akt/eNOS and Nrf2/ARE signaling.

Glycomimetics are a diverse array of saccharide-inspired compounds, designed to mimic the bioactive functions of glycosaminoglycans. Therefore, glycomimetics represent a unique source of novel therapies to target aberrant signaling and protein interactions in a wide range of diseases. We investigated the protective effects of four newly synthesized small molecule glycomimetics against lipid-induced endothelial dysfunction, with an emphasis on nitric oxide (NO) and oxidative stress.Four aromatic sugar mimetics were synthesized by the stepwise transformation of 2,5-dihydroxybenzoic to derivatives (C1-C4) incorporating sulfate groups to mimic the structure of heparan sulfate.Glycomimetic-treated human umbilical vein endothelial cells (HUVECs) were exposed to to model lipid-induced oxidative stress. Palmitate-induced impairment of NO production was restored by the glycomimetics, through activation of Akt/eNOS signaling. Furthermore, C1-C4 significantly inhibited palmitate-induced reactive species (ROS) production, lipid peroxidation, and activity and expression of NADPH oxidase. These effects were attributed to activation of the Nrf2/ARE pathway and downstream activation of cellular antioxidant and cytoprotective proteins. In ex vivo vascular reactivity studies, the glycomimetics (C1-C4) also demonstrated a significant improvement in endothelium-dependent relaxation and decreased ROS production and NADPH oxidase activity in isolated mouse thoracic aortic rings exposed to palmitate.The small molecule glycomimetics, C1-C4, protect against lipid-induced endothelial dysfunction through up-regulation of Akt/eNOS and Nrf2/ARE signaling pathways. Thus, carbohydrate-derived therapeutics are a new class of glycomimetic drugs targeting endothelial dysfunction, regarded as the first line of defense against vascular complications in cardiovascular disease.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: oxygen

Phycoremediation potential of marine microalga Tetraselmis indica on secondary treated domestic sewage for nutrient removal and biodiesel production.

Marine microalga Tetraselmis indica (T. indica) was cultivated in secondary treated domestic sewage (STDS) in batch mode. Optimization studies showed that after 14\xa0days of cultivation period, highest biomass yield reached was 0.88\xa0±\xa00.04\xa0g/L at the optimum temperature of 27\xa0±\xa01\xa0°C and light intensity of 135\xa0μmol\xa0m\xa0s. T. indica removed about 60.93% phosphate, 78.46% nitrate, 72.94% chemical demand (COD), 73.17% biological demand (BOD), 98.90% total dissolved solids (TDS) and heavy metals (83.11% Cd, 55.67% Ca, 45.12% Cu, 13.67% Mn, 50.88% Pb, and 98.92% Al) from STDS. The level of electrical conductivity was reduced to 0.0974\xa0±\xa00.045\xa0dS/m. The fatty methyl ester (FAME) profile showed the presence of (12.91%), oleic (35.94%), linoleic (14.89%) and eicosanoic (12.34%). This study indicates the potential of T. indica for removal of pollutants from STDS and also its capability of biodiesel production.

Keyword: oxygen

Resveratrol reduces intracellular reactive species levels by inducing autophagy through the AMPK-mTOR pathway.

Oxidative stress induced by free fatty aggravates endothelial injury, which leads to diabetic cardiovascular complications. Reduction of intracellular oxidative stress may attenuate these pathogenic processes. The dietary polyphenol resveratrol reportedly exerts potential protective effects against endothelial injury. This study determined whether resveratrol can reduce the (PA)-induced generation of reactive species (ROS) and further explored the underlying molecular mechanisms. We found that resveratrol significantly reduced the PA-induced endothelial ROS levels in human aortic endothelial cells. Resveratrol also induced endothelial cell autophagy, which mediated the effect of resveratrol on ROS reduction. Resveratrol stimulated autophagy via the AMP-activated protein kinase (AMPK)-mTOR pathway. Taken together, these data suggest that resveratrol prevents PA-induced intracellular ROS by autophagy regulation via the AMPK-mTOR pathway. Thus, the induction of autophagy by resveratrol may provide a novel therapeutic candidate for cardioprotection in metabolic syndrome.

Keyword: oxygen

Resistance exercise training and in\xa0vitro skeletal muscle oxidative capacity in older adults.

Whether resistance exercise training (RET) improves skeletal muscle substrate oxidative capacity and reduces mitochondrial production of reactive species in older adults remains unclear. To address this, 19 older males (≥60\xa0years) were randomized to a RET (n\xa0=\xa011) or to a waitlist control group (n\xa0=\xa08) that remained sedentary for 12\xa0weeks. RET was comprised of three upper body and four lower body movements on resistance machines. One set of 8-12 repetitions to failure of each movement was performed on three nonconsecutive days/week. Improvements in chest press and leg press strength were assessed using a three-repetition maximum (3 RM). Body composition was assessed via dual energy X-ray absorptiometry. Muscle biopsies were obtained from the vastus lateralis muscle at baseline and at both 3\xa0weeks and 12\xa0weeks. Palmitate and pyruvate oxidation rates were measured from the (14)CO2 produced from [1-(14)C] and [U-(14)C] pyruvate, respectively, during incubation of muscle homogenates. PGC-1α, TFAM, and PPARδ levels were quantified using qRT-PCR Citrate synthase (CS) and β-HAD activities were determined spectrophotometrically. Mitochondrial production of reactive species (ROS) were assessed using the Amplex Red Hydrogen Peroxide/Peroxidase assay. There were no significant changes in body weight or body composition following the intervention. Chest press and leg press strength (3RM) increased ~34% (both P\xa0<\xa00.01) with RET There were no significant changes in pyruvate or fatty oxidation or in the expression of target genes with the intervention. There was a modest increase (P\xa0<\xa00.05) in βHAD activity with RET at 12\xa0weeks but the change in CS enzyme activity was not significant. In addition, there were no significant changes in ROS production in either group following RET Taken together, the findings of this study suggest that 12\xa0weeks of low volume RET does not increase skeletal muscle oxidative capacity or reduce ROS production in older adults.© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Keyword: oxygen

Dynamics of ceramide generation and metabolism in response to fenretinide--Diversity within and among leukemia.

Fenretinide, N-(4-hydroxyphenyl)retinamide, (4-HPR), a synthetic retinoid, owes its cancer-toxic effects in part to the generation of ceramide, a potent tumor-suppressing sphingolipid. As such, 4-HPR has garnered considerable interest as a chemotherapeutic. Cancer cells, however, via various metabolic routes, inactivate ceramide, and this can limit 4-HPR efficacy. As relatively little is known regarding 4-HPR-induced ceramide management in acute myelogeneous leukemia (AML), we undertook the present study to evaluate the impact of 4-HPR on ceramide production, metabolism, and cytotoxicity. In KG-1, HL-60, and HL-60/VCR (multidrug resistant) human leukemia cells, 4-HPR induced 15-, 2-, and 20-fold increases in ceramide (measured using [3H]), respectively. By use of specific inhibitors we show that ceramide was produced by sphingomyelinase and de novo pathways in response to 4-HPR exposure. HL-60/VCR cells metabolized ceramide to glucosylceramide (GC). 4-HPR exposure (1.25-10 μM) reduced viability in all cell lines, with approximate IC50\'s ranging from 1 to 8.0 μM. Reactive species (ROS) were generated in response to 4-HPR treatment, and the concomitant cytotoxicity was reversed by addition of vitamin E. 4-HPR was not cytotoxic nor did it elicit ceramide formation in K562, a chronic myeloid leukemia cell line; however, K562 cells were sensitive to a cell-deliverable form of ceramide, C6-ceramide. Treatment of Molt-3, an acute lymphoblastic leukemia cell line, with 4-HPR revealed moderate ceramide production (5-fold over control), robust conversion of ceramide to GC and sphingomyelin, and resistance to 4-HPR and C6-ceramide. In conclusion, this work demonstrates diversity within and among leukemia in 4-HPR sensitivity and ceramide generation and subsequent metabolism. As such, knowledge of these metabolic pathways can provide guidance for enhancing ceramide-driven effects of 4-HPR in treatment of leukemia.Copyright © 2015 Elsevier Ltd. All rights reserved.

Keyword: oxygen

induces interleukin-1β secretion via NLRP3 inflammasomes and inflammatory responses through ROS production in human placental cells.

Maternal obesity, a major risk factor for adverse pregnancy complications, results in inflammatory cytokine release in the placenta. Levels of free fatty acids are elevated in the plasma of obese human. These fatty acids include obesity-related acids, which is a major saturated fatty , that promotes inflammatory responses. Increasing evidence indicates that nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) inflammasomes mediate inflammatory responses induced by endogenous danger signals. We hypothesized that inflammatory responses associated with gestational obesity cause inflammation. To test this hypothesis, we investigated the effect of on the activation of NLRP3 inflammasomes and inflammatory responses in a human Sw.71 trophoblast cell line. stimulated caspase-1 activation and markedly increased interleukin (IL)-1β secretion in Sw.71 cells. Treatment with a caspase-1 inhibitor diminished -induced IL-1β release. In addition, NLRP3 and caspase-1 genome editing using a CRISPR/Cas9 system in Sw.71 cells suppressed IL-1β secretion, which was stimulated by . Moreover, stimulated caspase-3 activation and inflammatory cytokine secretion (e.g., IL-6 and IL-8). -induced cytokine secretion were dependent on caspase-3 activation. In addition, -induced IL-1β, IL-6, and IL-8 secretion was depended on reactive species (ROS) generation. In conclusion, caused activation of NLRP3 inflammasomes and inflammatory responses, inducing IL-1β, IL-6, and IL-8 secretion, which is associated with ROS generation, in human Sw.71 placental cells. We suggest that obesity-related induces placental inflammation, resulting in association with pregnancy complications.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: oxygen

Application of quality-by-design approach to optimize diallyl disulfide-loaded solid lipid nanoparticles.

The current work was carried out by the principles of quality-by-design approach to develop an optimized solid lipid nanoparticles (SLNs) formulation of diallyl disulfide (DADS) through systematic statistical study. And its antitumor activity of DADS was also evaluated on breast cancer cell lines. To understand the effect of formulation variables (critical parameters) on the responses (critical quality attributes) of SLN, a 3-factor, 3-level Box-Behnken design, was explored to predict the responses such as particle size (Y1) and % entrapment efficiency (EE) (Y2) when concentration of surfactant (X1), amount of lipid (X2), and volume of solvent (X3) were selected as independent variables. Particle size analysis revealed that all the batches were within the nanometer range. DADS was released from the SLN much more rapidly at pH 4.5 than at pH 7.4, which is a desirable characteristic for tumor-targeted drug delivery. The cytotoxicity, reactive species (ROS), determination revealed that the antitumor activity of DADS is enhanced with SLN compared to DADS-free drug, and apoptosis is the mechanism underlying the cytotoxicity. The present study indicated the remarkable potential of DADS-SLN in enhancing the anticancer effect of DADS in breast cancer cells in vitro.

Keyword: oxygen

Homoplantaginin Inhibits -induced Endothelial Cells Inflammation by Suppressing TLR4 and NLRP3 Inflammasome.

(PA)-induced vascular endothelial inflammation plays a pivotal role in the occurrence and development of vascular diseases. The present study was conducted to examine the effect of homoplantaginin, a main flavonoid from a traditional Chinese medicine Salvia plebeia R. Br., on PA-treated human umbilical vein endothelial cells inflammation and the underlying molecular mechanism. Firstly, we found that homoplantaginin (0.1, 1, 10 μM) dose-dependently reduced expression of toll-like receptor-4 evoked by PA (100 μM). The inhibitory effect of homoplantaginin was further confirmed under lipopolysaccharide challenge. In addition, downstream adapted proteins including myeloid differentiation primary response gene 88, toll/interleukin-1 receptor-domain containing adaptor-inducing interferon-β and tumor necrosis factors receptor associated factor-6 were successfully inhibited by homoplantaginin under PA treatment. Also, we found that homoplantaginin tightly controlled PA-induced reactive species to prevent nucleotide-binding domain-like receptor 3 (NLRP3) inflammasome activation by suppressing reactive species-sensitive thioredoxin-interacting protein, NLRP3, and caspase-1. Meanwhile, protein and mRNA levels of inflammatory mediators (interleukin-1β, intercellular cell adhesion molecule-1, and monocyte chemotactic protein-1) were decreased by homoplantaginin. Furthermore, homoplantaginin restored PA-impaired nitric oxide generation. Taken together, these results indicated that homoplantaginin protected endothelial cells from ameliorating PA-induced endothelial inflammation via suppressing toll-like receptor-4 and NLRP3 pathways, and restoring nitric oxide generation, suggesting it may be a potential candidate for further development in the prevention and treatment of vascular diseases.

Keyword: oxygen

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less , indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: oxygen

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: oxygen

PINK1-Parkin-Mediated Mitophagy Protects Mitochondrial Integrity and Prevents Metabolic Stress-Induced Endothelial Injury.

Mitochondrial injury and dysfunction, a significant feature in metabolic syndrome, triggers endothelial cell dysfunction and cell death. Increasing evidence suggests that mitophagy, a process of autophagic turnover of damaged mitochondria, maintains mitochondrial integrity. PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase 1) and Parkin signaling is a key pathway in mitophagy control. In this study, we examined whether this pathway could protect mitochondria under metabolic stress. We found that (PA) induced significant mitophagy and activated PINK1 and Parkin in endothelial cells. Knocking down PINK1 or Parkin reduced mitophagy, leading to impaired clearance of damaged mitochondria and intracellular accumulation of mitochondrial fragments. Furthermore, PINK1 and Parkin prevented PA-induced mitochondrial dysfunction, ROS production and apoptosis. Finally, we show that PINK1 and Parkin were up-regulated in vascular wall of obese mice and diabetic mice. Our study demonstrates that PINK1-Parkin pathway is activated in response to metabolic stress. Through induction of mitophagy, this pathway protects mitochondrial integrity and prevents metabolic stress-induced endothelial injury.

Keyword: oxygen

Neuronal nitric oxide synthase modulation of intracellular Ca handling overrides fatty potentiation of cardiac inotropy in hypertensive rats.

Cardiac neuronal nitric oxide synthase (nNOS) is an important molecule that regulates intracellular Ca homeostasis and contractility of healthy and diseased hearts. Here, we examined the effects of nNOS on fatty (FA) regulation of left ventricular (LV) myocyte contraction in sham and angiotensin II (Ang II)-induced hypertensive (HTN) rats. Our results showed that (PA, 100\xa0μM) increased the amplitudes of sarcomere shortening and intracellular ATP in sham but not in HTN despite consumption rate (OCR) was increased by PA in both groups. Carnitine palmitoyltransferase-1 inhibitor, etomoxir (ETO), reduced OCR and ATP with PA in sham and HTN but prevented PA potentiation of sarcomere shortening only\xa0in sham. PA increased nNOS-derived NO only in HTN. Inhibition of nNOS with S-methyl-L-thiocitrulline (SMTC) prevented PA-induced OCR and restored PA potentiation of myocyte contraction in HTN. Mechanistically, PA increased intracellular Ca transient ([Ca]) without changing Ca influx via L-type Ca channel (I-) and reduced myofilament Ca sensitivity in sham. nNOS inhibition increased [Ca], I- and reduced myofilament Ca sensitivity prior to PA supplementation; as such, normalized PA increment of [Ca]. In HTN, PA reduced I- without affecting [Ca] or myofilament Ca sensitivity. However, PA increased I-, [Ca] and reduced myofilament Ca sensitivity following nNOS inhibition. Myocardial FA oxidation (F-fluoro-6-thia-heptadecanoic , F-FTHA) was comparable between groups, but nNOS inhibition increased it only in HTN. Collectively, PA increases myocyte contraction through stimulating [Ca] and mitochondrial activity in healthy hearts. PA-dependent cardiac inotropy was limited by nNOS in HTN, predominantly due to its modulatory effect on [Ca] handling.

Keyword: oxygen

Palmitoleic (16:1n7) increases consumption, fatty oxidation and ATP content in white adipocytes.

We have recently demonstrated that palmitoleic (16:1n7) increases lipolysis, glucose uptake and glucose utilization for energy production in white adipose cells. In the present study, we tested the hypothesis that palmitoleic modulates bioenergetic activity in white adipocytes.For this, 3\xa0T3-L1 pre-adipocytes were differentiated into mature adipocytes in the presence (or absence) of (16:0) or palmitoleic (16:1n7) at 100 or 200\xa0μM. The following parameters were evaluated: lipolysis, lipogenesis, fatty (FA) oxidation, ATP content, consumption, mitochondrial mass, citrate synthase activity and protein content of mitochondrial oxidative phosphorylation (OXPHOS) complexes.Treatment with 16:1n7 during 9\xa0days raised basal and isoproterenol-stimulated lipolysis, FA incorporation into triacylglycerol (TAG), FA oxidation, consumption, protein expression of subunits representing OXPHOS complex II, III, and V and intracellular ATP content. These effects were not observed in adipocytes treated with 16:0.Palmitoleic , by concerted action on lipolysis, FA esterification, mitochondrial FA oxidation, consumption and ATP content, does enhance white adipocyte energy expenditure and may act as local hormone.

Keyword: oxygen

Short-term starvation is a strategy to unravel the cellular capacity of oxidizing specific exogenous/endogenous substrates in mitochondria.

Mitochondrial oxidation of nutrients is tightly regulated in response to the cellular environment and changes in energy demands. studies evaluating the mitochondrial capacity of oxidizing different substrates are important for understanding metabolic shifts in physiological adaptations and pathological conditions, but may be influenced by the nutrients present in the culture medium or by the utilization of endogenous stores. One such influence is exemplified by the Crabtree effect (the glucose-mediated inhibition of mitochondrial respiration) as most experiments are performed in glucose-containing media. Here, using high-resolution respirometry, we evaluated the oxidation of endogenous or exogenous substrates by cell lines harboring different metabolic profiles. We found that a 1-h deprivation of the main energetic nutrients is an appropriate strategy to abolish interference of endogenous or undesirable exogenous substrates with the cellular capacity of oxidizing specific substrates, namely glutamine, pyruvate, glucose, or palmitate, in mitochondria. This approach primed mitochondria to immediately increase their consumption after the addition of the exogenous nutrients. All starved cells could oxidize exogenous glutamine, whereas the capacity for oxidizing palmitate was limited to human hepatocarcinoma Huh7 cells and to C2C12 mouse myoblasts that differentiated into myotubes. In the presence of exogenous glucose, starvation decreased the Crabtree effect in Huh7 and C2C12 cells and abrogated it in mouse neuroblastoma N2A cells. Interestingly, the fact that the Crabtree effect was observed only for mitochondrial basal respiration but not for the maximum respiratory capacity suggests it is not caused by a direct effect on the electron transport system.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: oxygen

Attenuation of -Induced Lipotoxicity by Chlorogenic through Activation of SIRT1 in Hepatocytes.

Saturated free fatty acids (FFAs) induce hepatocyte lipotoxicity, wherein oxidative stress-associated mitochondrial dysfunction is mechanistically involved. Chlorogenic (CGA), a potent antioxidant and anti-inflammatory compound, protects against high-fat-diet-induced oxidative stress and mitochondrial dysfunction in liver. This study investigates whether CGA protects against FFA-induced hepatocyte lipotoxicity via the regulation of mitochondrial fission/fusion and elucidates its underlying mechanisms.AML12 cell, a non-transformed hepatocyte cell line, is treated with palmitate. Here, it is shown that CGA prevents palmitate-induced lipotoxicity by activation of SIRT1 regulated mitochondrial morphology. CGA treatment mitigates oxidative stress and mitochondrial dysfunction, as evidenced by a decrease in reactive species (ROS) production, and an increase in mitochondrial mass and mitochondrial membrane potential. CGA also significantly decreases Bax expression and thereby reduces mitochondria-mediated caspase-dependent apoptosis. Mechanistically, CGA attenuates ROS-induced mitochondrial fragmentation by inhibiting dynamin-related protein 1 (Drp1) and enhancing Mfn2 expression. In contrast, the inhibitory effects of CGA on the generation of mitochondrial ROS and Drp1 are blocked by siRNA knockdown of SIRT1.Collectively, these findings show that supplementation with CGA protects hepatocytes from FFA-induced lipotoxicity through activation of SIRT1, which reverses the oxidative stress and dysfunction of mitochondrial biogenesis directly.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: oxygen

Effects of Grazing on the Behaviour, Oxidative and Immune Status, and Production of Organic Dairy Cows.

This study compared the effects of a short daily grazing time with those of permanent free-stall housing on the behaviour, oxidative status, immune response, and milk production of organically reared cows. During a 63-day period, two homogeneous groups of eight lactating Brown cows were allocated to either housing (H) in a free-stall building for 24 h/day. Feeding was based on a total mixed ration or grazing (G) on barley grass for 5 h/day, and housing in a free-stall structure with feeding was based on the same total mixed ration offered to the H group. With regard to behaviour, H cows spent more time idling, walking, drinking, and self-grooming, whereas G cows showed a greater intent to eat and interact socially. Moreover, G cows exhibited slightly higher reactive metabolites and similar biological antioxidant potential concentrations than the H group, which indicates that short grazing resulted in an almost negligible increase in oxidative stress and an unchanged antioxidant capacity. Skin tests, performed by injecting phytohemoagglutinin intradermally, indicated that G cows had thicker skin than H cows at the end of the trial, an index of a better cell-mediated immune response. Grazing did not affect milk yield but improved milk quality in terms of an increase in fat and a reduction in urea content, somatic cell count, and total microbial count. Milk from G cows was richer in saturated fatty acids, likely because of the contribution of present in the grazed barley grass, and also showed higher contents of some healthy fatty acids, such as rumenic and α-linolenic , and a lower omega-6/omega-3 ratio. These results show that including a short grazing time in the diets of organic dairy cows does not have negative consequences for milk production and contributes to improved milk quality as well as to a more efficient immune response in the cows.

Keyword: oxygen

Disruption of the mitochondria-associated ER membrane (MAM) plays a central role in -induced insulin resistance.

The mitochondria-associated ER membrane (MAM) is a specialized subdomain of ER that physically connects with mitochondria. Although disruption of inter-organellar crosstalk via the MAM impairs cellular homeostasis, its pathological significance in insulin resistance in type 2 diabetes mellitus remains unclear. Here, we reveal the importance of reduced MAM formation in the induction of fatty -evoked insulin resistance in hepatocytes. (PA) repressed insulin-stimulated Akt phosphorylation in HepG2 cells within 12h. Treatment with an inhibitor of the ER stress response failed to restore PA-mediated suppression of Akt activation. Mitochondrial reactive species (ROS) production did not increase in PA-treated cells. Even short-term exposure (3h) to PA reduced the calcium flux from ER to mitochondria, followed by a significant decrease in MAM contact area, suggesting that PA suppressed the functional interaction between ER and mitochondria. Forced expression of mitofusin-2, a critical component of the MAM, partially restored MAM contact area and ameliorated the PA-elicited suppression of insulin sensitivity with Ser473 phosphorylation of Akt selectively improved. These results suggest that loss of proximity between ER and mitochondria, but not perturbation of homeostasis in the two organelles individually, plays crucial roles in PA-evoked Akt inactivation in hepatic insulin resistance.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: oxygen

Attenuation of palmitate induced insulin resistance in muscle cells by harmala, clove and river red gum.

The present study aimed to decipher the mechanism of action of selected anti-diabetic plants extracts on mediated insulin resistance in muscle cells. Our results showed that extract from Peganum harmala seeds, Eucalyptus camaldulensis and Syzygium aromaticum leaves, showed significant antioxidant activity. We found that these extracts were able to affect stress signalling by reducing p-38 MAP kinase phosphorylation. They also reduced phosphorylation of substrate for insulin receptor (IRS) at serine residues and increased its phosphorylation at tyrosine residues and also enhanced PKB phosphorylation. Glucose uptake was also enhanced in muscle cells after treatment with these extracts. Extracts from Lantana camara, Psidium gujava fruit and different parts of Cassia alata did not affect FFA mediated down-regulation of insulin signalling. The study conclude that seeds of Peganum harmala and leaves of Eucalyptus camaldulensis and Syzygium aromaticum enhanced insulin signal transduction and glucose uptake in muscle cells via reducing oxidative stress. As a result, these herbal extracts may be considered useful to protect from insulin resistance.

Keyword: oxygen

Furanoid F- F6 Uniquely Induces NETosis Compared to C16 and C18 Fatty Acids in Human Neutrophils.

Various biomolecules induce neutrophil extracellular trap (NET) formation or NETosis. However, the effect of fatty acids on NETosis has not been clearly established. In this study, we focused on the NETosis-inducing ability of several lipid molecules. We extracted the lipid molecules present in Arabian Gulf catfish (, Val) skin gel, which has multiple therapeutic activities. Gas chromatography⁻mass spectrometry (GC-MS) analysis of the lipid fraction-3 from the gel with NETosis-inducing activity contained fatty acids including a furanoid F- (F6; 12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic ) and common long-chain fatty acids such as (PA; C16:0), palmitoleic (PO; C16:1), stearic (SA; C18:0), and oleic (OA; C18:1). Using pure molecules, we show that all of these fatty acids induce NETosis to different degrees in a dose-dependent fashion. Notably, F6 induces a unique form of NETosis that is rapid and induces reactive species (ROS) production by both NADPH oxidase (NOX) and mitochondria. F6 also induces citrullination of histone. By contrast, the common fatty acids (PA, PO, SA, and OA) only induce NOX-dependent NETosis. The activation of the kinases such as ERK (extracellular signal-regulated kinase) and JNK (c-Jun N-terminal kinase) is important for long-chain fatty -induced NETosis, whereas, in F--induced NETosis, Akt is additionally needed. Nevertheless, NETosis induced by all of these compounds requires the final chromatin decondensation step of transcriptional firing. These findings are useful for understanding F-- and other fatty -induced NETosis and to establish the active ingredients with therapeutic potential for regulating diseases involving NET formation.

Keyword: oxygen

Ablation of TMEM126B protects against heart injury via improving mitochondrial function in high fat diet (HFD)-induced mice.

The mitochondrial dysfunction in the pathogenesis of myocardial damage associated with high fat diet (HFD)-induced obesity remains largely unknown. Transmembrane protein 126B (TMEM126B), as a complex I assembly factor, plays a key role in regulating mitochondrial function. In the present study, the effects of TMEM126B on mitochondrial function were investigated using genetic knockout approach in HFD-induced mouse models with obesity. We found that TMEM126B was significantly increased in HFD-treated cardiac samples. Genetic ablation of TMEM126B alleviated HFD-mediated metabolic disorder and heart injury. TEM results suggested that cardiac mitochondrial integrity was improved in TMEM126B knockout mice compared with the wild type (WT) mice after HFD challenge. Additionally, the mitochondrial dysfunction induced by HFD was alleviated in mice with TMEM126B knockout, as evidenced by the decreased protein expression levels of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and increased expression of mitofusin-1 (MFN1). The mitochondrial impairments were further confirmed in (PA)-incubated cardiomyocytes, as evidenced by the down-regulated membrane potential and ATP levels, and by the up-regulated mitochondrial reactive species (ROS) production and DNA damage, which were significantly reversed by TMEM126B knockdown in\xa0vitro. Finally, TMEM126B ablation suppressed mitochondrial-dependent apoptotic death in the hearts of HFD mice. Therefore, TMEM126B led to mitochondrial impairments, contributing to the pathogenesis of HFD-induced cardiac injury, and blockage of TMEM126B could inhibit mitochondrial dysfunction, paving the road to new therapeutic modalities for the prevention of obesity-associated heart injury.Copyright © 2019. Published by Elsevier Inc.

Keyword: oxygen

Insight into the molecular mechanism of heme oxygenase-1 induction by docosahexaenoic in U937 cells.

Heme oxygenase-1 (HO-1) has anti-inflammatory effects on myeloid cells in response to various stimuli. To date, little is known about whether fatty acids can affect HO-1 induction. Here, we report the induction of HO-1 by docosahexaenoic (DHA) and the associated molecular mechanisms in human myelomonocytic lymphoma U937 cells. When U937 cells were treated with DHA, eicosapentaenoic , or oleic , DHA was the most effective inducer of HO-1. The activation of AKT and glycogen synthase kinase-3β did not significantly change after DHA treatment. However, DHA increased the activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), but not of other mitogen-activated protein kinases such as p38 and JNK. The increase in HO-1 expression was significantly inhibited by U0126, an ERK1/2 inhibitor. Nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf-2) and its binding to the HO-1 promoter significantly increased upon DHA treatment. An increase in intracellular reactive species was detected by dichlorofluorescein diacetate, but not by hydroethidium or 2-[6-(4-hydroxy)phenoxy-3H-xanthen-3-on-9-yl] benzoic after DHA treatment. Pretreatment with NAC dramatically inhibited the ERK1/2 activation, binding of Nrf-2 to antioxidant response elements (AREs) located in the HO-1 promoter and the induction of HO-1 by DHA. In conclusion, DHA increased HO-1 expression in U937 cells via activation of ERK1/2 and increased Nrf-2 binding to ARE in the HO-1 promoter. These findings will help develop better strategies for treating inflammatory disorders with DHA.Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Keyword: oxygen

Withaferin A protects against -induced endothelial insulin resistance and dysfunction through suppression of oxidative stress and inflammation.

Activation of inflammatory pathways via reactive species (ROS) by free fatty acids (FFA) in obesity gives rise to insulin resistance and endothelial dysfunction. Withaferin A (WA), possesses both antioxidant and anti-inflammatory properties and therefore would be a good strategy to suppress (PA)-induced oxidative stress and inflammation and hence, insulin resistance and dysfunction in the endothelium. Effect of WA on PA-induced insulin resistance in human umbilical vein endothelial cells (HUVECs) was determined by evaluating insulin signaling mechanisms whilst effect of this drug on PA-induced endothelial dysfunction was determined in acetylcholine-mediated relaxation in isolated rat aortic preparations. WA significantly inhibited ROS production and inflammation induced by PA. Furthermore, WA significantly decreased TNF-α and IL-6 production in endothelial cells by specifically suppressing IKKβ/NF-κβ phosphorylation. WA inhibited inflammation-stimulated IRS-1 serine phosphorylation and improved the impaired insulin PI3-K signaling, and restored the decreased nitric oxide (NO) production triggered by PA. WA also decreased endothelin-1 and plasminogen activator inhibitor type-1 levels, and restored the impaired endothelium-mediated vasodilation in isolated aortic preparations. These findings suggest that WA inhibited both ROS production and inflammation to restore impaired insulin resistance in cultured endothelial cells and improve endothelial dysfunction in rat aortic rings.

Keyword: oxygen

Exercise Training Protects Cardiomyocytes from Deleterious Effects of Palmitate.

We investigated the effects of palmitate, a high saturated fat, on Ca, action potential and reactive species dynamics in cardiomyocytes from untrained and trained mice. Male mice were subjected to moderate intensity exercise training on a treadmill. Cardiomyocytes of untrained and trained mice were isolated, treated for 30\u2009min with palmitate and intracellular calcium transient and action potential duration were recorded. Additionally, we assessed reactive species generation. Treatment of cardiomyocytes from untrained mice with palmitate induced a significant decrease in Ca transient magnitude by 34%. Exercise training did not change cardiomyocyte Ca dynamics in the control group. However, trained cardiomyocytes were protected from deleterious effects of palmitate. Action potential duration was not altered by palmitate in either untrained or trained cardiomyocytes. Moreover, palmitate treatment increased reactive species generation in both untrained and trained cardiomyocytes. Nevertheless, the levels of reactive species in trained cardiomyocytes treated with palmitate were still 27% lower than those seen at basal conditions in untrained cardiomyocytes. Taken together, these findings indicate that exercise training protects cardiomyocytes from deleterious effects of palmitate possibly by inhibiting exacerbated ROS production.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: oxygen

Effect of salinity stress on growth, lipid productivity, fatty composition, and biodiesel properties in Acutodesmus obliquus and Chlorella vulgaris.

Two microalgae strains including Chlorella vulgaris and Acutodesmus obliquus were grown on BG11 medium with salinity stress ranging from 0.06 to 0.4\xa0M NaCl. Highest lipid content in C. vulgaris and A. obliquus was 49 and 43% in BG11 amended with 0.4\xa0M NaCl. The microalgal strains C. vulgaris and A. obliquus grow better at 0.06\xa0M NaCl concentration than control condition. At 0.06\xa0M NaCl, improved dry biomass content in C. vulgaris and A. obliquus was 0.92 and 0.68\xa0gL, respectively. Stress biomarkers like reactive species, antioxidant enzyme catalase, and ascorbate peroxidase were also lowest at 0.06\xa0M NaCl concentration revealing that both the microalgal strains are well acclimatized at 0.06\xa0M NaCl concentration. The fatty composition of the investigated microalgal strains was also improved by increased NaCl concentration. At 0.4\xa0M NaCl, (37%), oleic (15.5%), and linoleic (20%) were the dominant fatty acids in C. vulgaris while (54%) and stearic (26.6%) were major fatty acids found in A. obliquus. Fatty profiling of C. vulgaris and A. obliquus significantly varied with salinity concentration. Therefore, the study showed that salt stress is an effective stress that could increase not only the lipid content but also improved the fatty composition which could make C. vulgaris and A. obliquus potential strains for biodiesel production.

Keyword: oxygen

Neutral ceramidase-enriched exosomes prevent -induced insulin resistance in H4IIEC3 hepatocytes.

Oversupply of free fatty acids such as (PA) from the portal vein may cause liver insulin resistance. Production of reactive species plays a pivotal role in PA-induced insulin resistance in H4IIEC3 hepatocytes. Recently, we found that exosomes secreted from INS-1 cells that were transfected with neutral ceramidase (NCDase) plasmids had raised NCDase activity; these NCDase-enriched exosomes could inhibit PA-induced INS-1 cell apoptosis. Here, we showed that PA reduced insulin-stimulated tyrosine phosphorylation of insulin receptor substrate 2 and decreased insulin-stimulated uptake of the fluorescent glucose analog 2-NBDG, confirming that insulin resistance occurred in PA-treated H4IIEC3 cells. Moreover, NCDase-enriched exosomes from INS-1 cells rescued PA-induced H4IIEC3 insulin resistance and blocked PA-induced reactive species production in which ceramide was involved.

Keyword: oxygen

Hepatocyte miR-33a mediates mitochondrial dysfunction and hepatosteatosis by suppressing NDUFA5.

Emerging evidence suggests that microRNAs (miRNAs) are essential for metabolic haemostasis of liver tissues. Among them, miR-33a is supposed to modulate the cholesterol export and fatty oxidation, but whether miR-33a involves in the process of fatty liver disease is unclear. To disclose the hypothesis, we utilized miR-33a mimic and antisense to explore their effects in primary hepatocytes or high-fat diet (HFD)-fed mice. Treatment with (PA) or HFD significantly increased the expression of miR-33a in hepatocytes or liver tissues. In primary hepatocytes, miR-33a mimic decreased mitochondrial function, including reduction of ATP production and consumption, whereas miR-33a inhibition protected PA-induced mitochondrial dysfunction. Interestingly, miR-33a selectively suppressed mitochondrial complex I activity and protein expression, but not other complexes. Through bioinformatics prediction, we found miR-33a directly targeted on the 3\'-UTR of NDUFA5. Dual-luciferase reporter analysis further confirmed the direct suppression of miR-33a on NDUFA5 expression. More importantly, administration of miR-33a antisense could effectively restore HFD-induced mitochondrial dysfunction through up-regulation of NDUFA5 levels. Mice treated with miR-33a antisense also exhibited improved liver function and structural disorders under obese status. Taken together, miR-33a was an important mediator of hepatocyte mitochondrial function, and the therapeutic benefits implied miR-33a antisense had the potential clinical application in combating the fatty liver disease.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: oxygen

AIF loss deregulates hematopoiesis and reveals different adaptive metabolic responses in bone marrow cells and thymocytes.

Mitochondrial metabolism is a tightly regulated process that plays a central role throughout the lifespan of hematopoietic cells. Herein, we analyze the consequences of the mitochondrial oxidative phosphorylation (OXPHOS)/metabolism disorder associated with the cell-specific hematopoietic ablation of apoptosis-inducing factor (AIF). AIF-null (AIF ) mice developed pancytopenia that was associated with hypocellular bone marrow (BM) and thymus atrophy. Although myeloid cells were relatively spared, the B-cell and erythroid lineages were altered with increased frequencies of precursor B cells, pro-erythroblasts I, and basophilic erythroblasts II. T-cell populations were dramatically reduced with a thymopoiesis blockade at a double negative (DN) immature state, with DN1 accumulation and delayed DN2/DN3 and DN3/DN4 transitions. In BM cells, the OXPHOS/metabolism dysfunction provoked by the loss of AIF was counterbalanced by the augmentation of the mitochondrial biogenesis and a shift towards anaerobic glycolysis. Nevertheless, in a caspase-independent process, the resulting excess of reactive species compromised the viability of the hematopoietic stem cells (HSC) and progenitors. This led to the progressive exhaustion of the HSC pool, a reduced capacity of the BM progenitors to differentiate into colonies in methylcellulose assays, and the absence of cell-autonomous HSC repopulating potential in vivo. In contrast to BM cells, AIF thymocytes compensated for the OXPHOS breakdown by enhancing fatty β-oxidation. By over-expressing CPT1, ACADL and PDK4, three key enzymes facilitating fatty β-oxidation (e.g., assimilation), the AIF thymocytes retrieved the ATP levels of the AIF cells. As a consequence, it was possible to significantly reestablish AIF thymopoiesis in vivo by feeding the animals with a high-fat diet complemented with an antioxidant. Overall, our data reveal that the mitochondrial signals regulated by AIF are critical to hematopoietic decision-making. Emerging as a link between mitochondrial metabolism and hematopoietic cell fate, AIF-mediated OXPHOS regulation represents a target for the development of new immunomodulatory therapeutics.

Keyword: oxygen

Dihydromyricetin Ameliorates Nonalcoholic Fatty Liver Disease by Improving Mitochondrial Respiratory Capacity and Redox Homeostasis Through Modulation of SIRT3 Signaling.

Our previous clinical trial indicated that the flavonoid dihydromyricetin (DHM) could improve hepatic steatosis in patients with nonalcoholic fatty liver disease (NAFLD), altough the potential mechanisms of these effects remained elusive. Here, we investigated the hepatoprotective role of DHM on high-fat diet (HFD)-induced NAFLD. DHM supplementation could effectively ameliorate the development of NAFLD by inhibiting hepatic lipid accumulation both in HFD-fed wild-type mice and in -induced hepatocytes. We reveal for the first time that mitochondrial dysfunction characterized by ATP depletion and augmented oxidative stress could be reversed by DHM treatment. Moreover, DHM enhanced the mitochondrial respiratory capacity by increasing the expression and enzymatic activities of mitochondrial complexes and increased mitochondrial reactive species scavenging by restoring manganese superoxide dismutase (SOD2) activity. Interestingly, the benefits of DHM were abrogated in knockout (SIRT3KO) mice and in hepatocytes transfected with siRNA or treated with an SIRT3-specific inhibitor. We further showed that DHM could increase SIRT3 expression by activating the adenosine monophosphate-activated protein kinase (AMPK)-peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC1α)/estrogen-related receptor-α (ERRα) signaling pathway. Our work indicates that SIRT3 plays a critical role in the DHM-mediated beneficial effects that include ameliorating mitochondrial dysfunction and oxidative stress in a nutritional NAFLD model both and Our results suggest that DHM prevents NAFLD by improving mitochondrial respiratory capacity and redox homeostasis in hepatocytes through a SIRT3-dependent mechanism. These results could provide a foundation to identify new DHM-based preventive and therapeutic strategies for NAFLD.

Keyword: oxygen

Engineering cytosolic acetyl-coenzyme A supply in Saccharomyces cerevisiae: Pathway stoichiometry, free-energy conservation and redox-cofactor balancing.

Saccharomyces cerevisiae is an important industrial cell factory and an attractive experimental model for evaluating novel metabolic engineering strategies. Many current and potential products of this yeast require acetyl coenzyme A (acetyl-CoA) as a precursor and pathways towards these products are generally expressed in its cytosol. The native S. cerevisiae pathway for production of cytosolic acetyl-CoA consumes 2 ATP equivalents in the acetyl-CoA synthetase reaction. Catabolism of additional sugar substrate, which may be required to generate this ATP, negatively affects product yields. Here, we review alternative pathways that can be engineered into yeast to optimize supply of cytosolic acetyl-CoA as a precursor for product formation. Particular attention is paid to reaction stoichiometry, free-energy conservation and redox-cofactor balancing of alternative pathways for acetyl-CoA synthesis from glucose. A theoretical analysis of maximally attainable yields on glucose of four compounds (n-butanol, citric , and farnesene) showed a strong product dependency of the optimal pathway configuration for acetyl-CoA synthesis. Moreover, this analysis showed that combination of different acetyl-CoA production pathways may be required to achieve optimal product yields. This review underlines that an integral analysis of energy coupling and redox-cofactor balancing in precursor-supply and product-formation pathways is crucial for the design of efficient cell factories.Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: oxygen

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the liver of high-fat and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in liver sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Keyword: oxygen

Blueberry Metabolites Attenuate Lipotoxicity-Induced Endothelial Dysfunction.

Lipotoxicity-induced endothelial dysfunction is an important vascular complication associated with diabetes. Clinical studies support the vascular benefits of blueberry anthocyanins, but the underlying mechanism is unclear. The hypothesis that metabolites of blueberry anthocyanins attenuate lipotoxicity-induced endothelial dysfunction was tested.Human aortic endothelial cells (HAECs) were treated for 6 h with either: (i) the parent anthocyanins (malvidin-3-glucoside and cyanidin-3-glucoside); or (ii) the blueberry metabolites (hydroxyhippuric , hippuric , benzoic -4-sulfate, isovanillic -3-sulfate, and vanillic -4-sulfate), at concentrations known to circulate in humans following blueberry consumption. For the last 5 h HAECs were treated with palmitate or vehicle. HAECs treated with palmitate displayed elevated reactive species generation, increased mRNA expression of NOX4, chemokines, adhesion molecules, and IκBα, exaggerated monocyte binding, and suppressed nitric oxide production. Of note, the damaging effects of palmitate were ameliorated in HAECs treated with blueberry metabolites but not parent anthocyanins. Further, important translational relevance of these results was provided by our observation that palmitate-induced endothelial dysfunction was lessened in arterial segments that incubated concurrently with blueberry metabolites.The presented findings indicate that the vascular benefits of blueberry anthocyanins are mediated by their metabolites. Blueberries might complement existing therapies to lessen vascular complications.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: oxygen

PINK1 alleviates palmitate induced insulin resistance in HepG2 cells by suppressing ROS mediated MAPK pathways.

Oxidative stress is an important pathogenesis of insulin resistance (IR) and Type 2 diabetes mellitus (T2DM). Studies have shown that knockdown of PTEN-induced putative kinase 1 (PINK1) causes oxidative stress and mitophagy. In db/db mice, PINK1 protein level is down-regulated. However, little is known regarding the mechanism by which PINK1 modulates IR in response to reactive species (ROS) induced stress. In our study, PINK1 expression decreased during palmitate (PA) induced IR in HepG2 cells and the hepatic tissues of high fat diet (HFD) fed mice. Additionally, free fatty acids (FFAs) could increase ROS and suppress insulin signaling pathway, which was indicated by reduced phosphorylation of protein kinase B (AKT) and glycogen synthase kinase 3β (GSK-3β). In addition, insulin induced glucose uptake decreased and the expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase), two key gluconeogenic enzymes, was up-regulated after PA treatment. Intriguingly, PINK1 overexpression could lead to opposite results. Moreover, PA induced hepatic IR through C-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) pathways, which were rescued by PINK1 overexpression. In summary, our results demonstrate that PINK1 promoted hepatic IR via JNK and ERK pathway in PA treated HepG2 cells, implying a novel molecular target for the therapy of diabetes.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: oxygen

Synergistic effects of metformin with liraglutide against endothelial dysfunction through GLP-1 receptor and PKA signalling pathway.

Metformin or glucagon-like peptide-1 (GLP-1) analogue liraglutide has cardiovascular benefits. However, it is not clear whether their combined treatment have additive or synergistic effects on the vasculature. In this study, human umbilical vein endothelial cells (HUVECs), exposed to (PA) to induce endothelial dysfunction, were incubated with metformin, liraglutide or their combination. High fat diet (HFD)-fed ApoE mice were randomized into control, metformin, liraglutide, and combination treatment groups. Results showed that in PA-treated HUVECs and HFD-fed ApoE mice, combination of metformin and liraglutide at lower dose significantly improved endothelial dysfunction compared with the single treatment. Metformin upregulated GLP-1 receptor (GLP-1R) level and protein kinase A (PKA) phosphorylation. However, PKA inhibition but not GLP-1R blockade eliminated the protective effects of metformin on endothelial function. Furthermore, AMPK inhibitor compound C abolished the metformin-mediated upregulation of GLP-1R level and PKA phosphorylation. In conclusion, combination of metformin and liraglutide has synergistic protective effects on endothelial function. Moreover, metformin stimulates GLP-1R and PKA signalling via AMPK-dependent pathway, which may account for its synergistic protective effects with liraglutide. Our findings provide new insights on the interaction between metformin and GLP-1, and provide important information for designing new GLP-1-based therapy strategies in treating type 2 diabetes.

Keyword: oxygen

Fatty Metabolism is Associated With Disease Severity After H7N9 Infection.

Human infections with the H7N9 virus could lead to lung damage and even multiple organ failure, which is closely associated with a high mortality rate. However, the metabolic basis of such systemic alterations remains unknown.This study included hospitalized patients (n\u202f=\u202f4) with laboratory-confirmed H7N9 infection, healthy controls (n\u202f=\u202f9), and two disease control groups comprising patients with pneumonia (n\u202f=\u202f9) and patients with pneumonia who received steroid treatment (n\u202f=\u202f10). One H7N9-infected patient underwent lung biopsy for histopathological analysis and expression analysis of genes associated with lung homeostasis. H7N9-induced systemic alterations were investigated using metabolomic analysis of sera collected from the four patients by using ultra-performance liquid chromatography-mass spectrometry. Chest digital radiography and laboratory tests were also conducted.Two of the four patients did not survive the clinical treatments with antiviral medication, steroids, and therapy. Biopsy revealed disrupted expression of genes associated with lung epithelial integrity. Histopathological analysis demonstrated severe lung inflammation after H7N9 infection. Metabolomic analysis indicated that fatty metabolism may be inhibited during H7N9 infection. Serum levels of , erucic , and phytal may negatively correlate with the extent of lung inflammation after H7N9 infection. The changes in fatty levels may not be due to steroid treatment or pneumonia.Altered structural and secretory properties of the lung epithelium may be associated with the severity of H7N9-infection-induced lung disease. Moreover, fatty metabolism level may predict a fatal outcome after H7N9 virus infection.Copyright © 2018. Published by Elsevier B.V.

Keyword: oxygen

Yeast Mpo1 Is a Novel Dioxygenase That Catalyzes the α-Oxidation of a 2-Hydroxy Fatty in an Fe-Dependent Manner.

Phytosphingosine (PHS) is the major long-chain base component of sphingolipids in The PHS metabolic pathway includes a fatty (FA) α-oxidation reaction. Recently, we identified the novel protein Mpo1, which is involved in PHS metabolism. However, the details of the FA α-oxidation reaction and the role of Mpo1 in PHS metabolism remained unclear. In the present study, we revealed that Mpo1 is involved in the α-oxidation of 2-hydroxy (2-OH) (C-COOH) in the PHS metabolic pathway. Our assay revealed that not only the Mpo1-containing membrane fraction but also the soluble fraction was required for the α-oxidation of 2-OH C-COOH. The addition of Fe eliminated the need for the soluble fraction. Purified Mpo1 converted 2-OH C-COOH to C-COOH in the presence of Fe, indicating that Mpo1 is the enzyme body responsible for catalyzing the FA α-oxidation reaction. This reaction was also found to require an molecule. Our findings indicate that Mpo1 catalyzes the FA α-oxidation reaction as 2-OH fatty dioxygenase, mediated by iron(IV) peroxide. Although numerous Mpo1 homologs exist in bacteria, fungi, protozoa, and plants, their functions had not yet been clarified. However, our findings suggest that these family members function as dioxygenases.Copyright © 2019 American Society for Microbiology.

Keyword: oxygen

Exploring the entry route of and palmitoylcarnitine into myoglobin.

Myoglobin, besides its role in turnover, has gained recognition as a potential regulator of lipid metabolism. Previously, we confirmed the interaction of fatty acids and acylcarnitines with Oxy-Myoglobin, using both molecular dynamic simulations and Isothermal Titration Calorimetry studies. However, those studies were limited to testing only the binding sites derived from homology to fatty binding proteins and predictions using automated docking. To explore the entry mechanisms of the lipid ligands into myoglobin, we conducted molecular dynamic simulations of murine Oxy- and Deoxy-Mb structures with palmitate or palmitoylcarnitine starting at different positions near the protein surface. The simulations indicated that both ligands readily (under ∼10-20 ns) enter the Oxy-Mb structure through a dynamic area ("portal region") near heme, known to be the entry point for small molecule gaseous ligands like O, CO and NO. The entry is not observed with Deoxy-Mb where lipid ligands move away from protein surface, due to a compaction of the entry portal and the heme-containing crevice in the Mb protein upon O removal. The results suggest quick spontaneous binding of lipids to Mb driven by hydrophobic interactions, strongly enhanced by oxygenation, and consistent with the emergent role of Mb in lipid metabolism.Published by Elsevier Inc.

Keyword: oxygen

Up-regulation of HO-1 by Nrf2 activation protects against -induced ROS increase in human neuroblastoma BE(2)-M17 cells.

Saturated fatty acids (SFAs) induce reactive species (ROS) production in neurons. Extracellular signal regulated kinase (ERK)/nuclear factor erythroid-2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) is a ROS response pathway. Therefore, high ROS is always accompanied by increase of HO-1, an anti-oxidative enzyme; but it remains unknown why there is no significant reduction of ROS with the increase of HO-1 in SFAs-treated neurons. We hypothesized that the up-regulation of HO-1 is compensatory for response to fatty -induced oxidative stress but not enough to reduce ROS levels. We evaluated the anti-ROS effect of HO-1 and the involved pathway in (PA)-treated human neuroblastoma BE(2)-M17 cells. As expected, PA-induced ROS increase was accompanied by activation of the ERK-Nrf2-HO-1 pathway, as demonstrated by an increase in ERK phosphorylation, Nrf2 phosphorylation and nuclear accumulation, and HO-1 expression at the mRNA and protein levels, in a PA-dose-dependent manner. In contrast, administration of the ROS scavenger NAC significantly reduced the levels of PA-regulated ROS and HO-1 protein. However, the ERK inhibitor U0126 not only reversed the activating effect of PA on the ERK-Nrf2-HO-1 pathway but also aggravated PA-induced ROS. Furthermore, the Nrf2-specific activator NK-252 significantly increased PA-up-regulated HO-1 protein and alleviated PA-induced ROS. Therefore, our results suggest that up-regulation of HO-1 in PA-treated neurons is a compensatory response to ROS increase and that increasing HO-1 expression by Nrf2 activation can prevent the process of ROS production in PA-treated neurons.Copyright © 2018. Published by Elsevier Inc.

Keyword: oxygen

Effect of Hyperbaric Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

Metabolic changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: oxygen

Astragalosides IV protected the renal tubular epithelial cells from free fatty acids-induced injury by reducing oxidative stress and apoptosis.

Renal tubular injury is associated with the development of diabetic nephropathy (DN) and the end-stage renal disease (ESRD). Free fatty acids (FFAs)-associated lipotoxicity contributes to injury of proximal renal tubular epithelial (HK-2) cells in diabetes. (PA) which is the most abundant saturated fatty in FFAs is closely associated with the gradual decline of renal function. Astragalosides IV (AS-IV) has a variety of pharmacological effects such as anti-inflammation and anti-oxidation. In the current study, we investigated the effects of AS-IV on PA-induced apoptosis of HK-2 cells and the underlying mechanisms. The results showed that AS-IV (10, 20, 40\u2009μmol/L) could alleviate PA-induced apoptosis of HK-2 cells. We found that AS-IV reduced the expression of Bax and cleaved-caspase3, but increased the expression of Bcl-2 and phosphorylated Nrf2 in HK-2 cells. Moreover, AS-IV reduced the level of reactive species (ROS) in the cells. Our study suggests that AS-IV could protect against PA-induced apoptosis in HK-2 cells by inhibiting ROS generation and apoptotic protein expression. This study may provide a new theoretical option for the patients with type 2 diabetes.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: oxygen

Chemical Composition and Antioxidant Activity of Monguba (Pachira aquatica) Seeds.

Monguba fruit has a seed with a chestnut-like flavor that can be consumed boiled, fried, and roasted. These nutritious seeds also have been used in popular medicine to treat several diseases. Nevertheless, the nutritional and functional potential of monguba seed is still underexploited. In this sense, we investigated the nutritional and functional components of monguba seeds. These seeds showed high total content of sugars, mainly sucrose, whereas the content of the raffinose family oligosaccharides was low. The mineral assay showed high amount of minerals, namely potassium, calcium, magnesium and zinc, which indicate that monguba seeds can be a new source of these minerals. UHPLC-ESI-MS/MS analysis showed caffeic, ferulic and 4-hydroxybenzoic acids as the main phenolic compounds, mainly in the esterified form, in these seeds. Monguba seed showed high lipid content, in which the main compounds were and γ-tocopherol. The soluble and insoluble phenolic fractions from monguba seeds showed high antioxidant activity measured by the radical absorption capacity (ORAC) and the trolox equivalent antioxidant capacity (TEAC) assays. Therefore, the monguba seeds have great potential to be explored by food, pharmaceutical and cosmetic industries due to their chemical composition.Copyright © 2019. Published by Elsevier Ltd.

Keyword: oxygen

Characterization of aerobic oil and grease-degrading bacteria in wastewater.

A bacterial consortium that degrades cooking oil (CO) has been isolated in wastewater (WW) samples, by enrichment in olive CO. This consortium could degrade 90% of CO within 7-9 days (from an initial 1% [w/v]), and it is more active at alkaline conditions. The 16S ribonucleic (RNA) gene analysis showed that it contains five bacterium species: Stenotrophomonas rhizophila, Sphingobacterium sp., Pseudomonas libanensis, Pseudomonas poae and Pseudomonas aeruginosa. This consortium can degrade the free fatty acids (FFA): , stearic, oleic, linoleic and linolenic acids; glycerol, glucose and amylose; and albumin, but could not efficiently degrade carboxymethyl-cellulose. Each strain could also degrade CO and FFAs. The level of bacterial crude-activity of extracellular lipases was found to be between 0.2 and 4U/ml. Using synthetic WW, the consortium could reduce 80% of the chemical demand [from 10550\u2009±\u20092828\u2005mg/l], 80% of nitrogen (from 410\u2009±\u200978\u2009mgl/l) and 57% of phosphorus (from 93\u2009±\u200925\u2005mg/l). Thus, this consortium can be utilized in the removal of CO from WW.

Keyword: oxygen

Removal of nutrients and organic pollution load from pulp and paper mill effluent by microalgae in outdoor open pond.

A mixed culture of microalgae, containing two Scenedesmus species, was analysed to determine its potential in coupling of pulp and paper mill effluent treatment and microalgal cultivation. Laboratory studies suggested that 60% concentration of wastewater was optimum for microalgal cultivation. A maximum of 82% and 75% removal of BOD and COD respectively was achieved with microalgal cultivation in outdoor open pond. By the end of the cultivation period, 65% removal of NO3-N and 71.29% removal of PO4-P was observed. The fatty composition of mixed microalgal culture cultivated with effluent showed the , oleic , linoleic and α-linolenic as major fatty acids. The results obtained suggest that pulp and paper mill effluent could be used effectively for cultivation of microalgae to minimise the freshwater and nutrient requirements.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty composition.

A profound analysis of A.\xa0tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty profile. Plants exposed to 125-250\xa0mm NaCl showed higher photochemical light harvesting efficiencies and lower energy dissipation rates. under higher NaCl exposure, there is evident damage of the evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the energy fluxes entering the PS II, in spite of the high amounts of energy dissipated and the loss of PS II antennae connectivity. Energy dissipation was mainly achieved through the auroxanthin pathway. Total fatty content displayed a similar trend, being also higher under 125-250\xa0mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic and allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250\xa0mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of energy dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.© 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Keyword: oxygen

Water Extract of L. Ameliorates Non-Alcoholic Fatty Liver Disease.

Our aim was to investigate whether hot water extract (CLW) of L. could prevent non-alcoholic fatty liver disease (NAFLD). HepG2 cells were treated with free fatty (FFA) mixture (oleic : , 2:1) for 24 h to stimulate in vitro fatty liver. In addition, C57BL/6 mice were fed 60 kcal% high-fat (HF) diet for eight weeks to induce fatty liver in vivo. Intracellular reactive species (ROS) and malondialdehyde (MDA) productions were increased by FFA and HF-diet, but supplementation with CLW significantly decreased these levels. CLW treatment ameliorated antioxidant activities that were suppressed by exposure to the FFA and HF-diet. Cluster of differentiation 36 (CD36) and fatty transport proteins (FATP2 and FATP5) were increased in HF-diet groups, while CLW suppressed their expression levels. Moreover, sterol regulatory element-binding protein-1c (SREBP-1c), acetyl-coenzyme A carboxylase (ACC), and fatty synthase (FAS) expression levels were down-regulated in the CLW groups compared to HF-diet groups. On the other hand, 5\' adenosine monophosphate-activated protein kinase (AMPK), Peroxisome proliferator-activated receptor alpha (PPAR-α), and carnitine palmitoyltransferase 1 (CPT-1) expressions were up-regulated in the CLW groups. HF-diet fed mice showed high hepatic triglycerides (TG) content compared to the normal diet mice. However, the administration of CLW restored the hepatic TG level, indicating an inhibitory effect against lipid accumulation by CLW. These results suggest that CLW could be a potentially useful agent for the prevention of NAFLD through modulating fatty uptake.

Keyword: oxygen

Macrophages with a deletion of the () gene have a more proinflammatory phenotype.

Phosphoenolpyruvate carboxykinase (Pck1) is a metabolic enzyme that is integral to the gluconeogenic and glyceroneogenic pathways. However, Pck1\'s role in macrophage metabolism and function is unknown. Using stable isotopomer MS analysis in a mouse model with a myeloid cell-specific deletion, we show here that this deletion increases the proinflammatory phenotype in macrophages. Incubation of LPS-stimulated bone marrow-derived macrophages (BMDM) with [U-C]glucose revealed reduced C labeling of citrate and malate and increased C labeling of lactate in Pck1-deleted bone marrow-derived macrophages. We also found that the Pck1 deletion in the myeloid cells increases reactive species (ROS). Of note, this altered macrophage metabolism increased expression of the M1 cytokines TNFα, IL-1β, and IL-6. We therefore conclude that contributes to M1 polarization in macrophages. Our findings provide important insights into the factors determining the macrophage inflammatory response and indicate that Pck1 activity contributes to metabolic reprogramming and polarization in macrophages.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: oxygen

Vanillic alleviates -induced oxidative stress in human umbilical vein endothelial cells via Adenosine Monophosphate-Activated Protein Kinase signaling pathway.

Vanillic (VA), one of the phenolic acids metabolized by anthocyanidins, can modulate vascular reactivity by reducing the superoxide. We investigated that VA alleviated fatty -induced oxidative stress and clarified its potential mechanisms in human umbilical vein endothelial cells (HUVECs). Our results showed that VA reduced the production of reactive species and malondialdehyde levels. It also restored mitochondrial membrane potential and enhanced the activities of antioxidant enzymes. In addition, VA promoted the expression of p-Nrf2 and HO-1 through LKB1/AMPK signaling pathway, as well as the level of SIRT1 and PGC-1α. Moreover, compound C reduced the effect of VA on the enhancement of p-Nrf2 and HO-1. These results indicated that AMPK was an important target molecule of VA in the process of alleviating oxidative stress in HUVECs, providing a new potential evidence for vascular protection of anthocyanin in vitro. PRACTICAL APPLICATIONS: As a phenolic derivative and phase II metabolite of anthocyanins in vivo, VA can be found in various edible plants and fruits. This study revealed that VA improved oxidative stress in endothelial cells stimulated by by activating AMPK and its downstream proteins. VA could be a potential functional material for the protection of diabetic vascular complications.© 2019 Wiley Periodicals, Inc.

Keyword: oxygen

Resveratrol Ameliorates Palmitate-Induced Inflammation in Skeletal Muscle Cells by Attenuating Oxidative Stress and JNK/NF-κB Pathway in a SIRT1-Independent Mechanism.

Resveratrol has been shown to exert anti-inflammatory and anti-oxidant effects in a variety of cell types, however, its role in prevention of inflammatory responses mediated by palmitate in skeletal muscle cells remains unexplored. In the present study, we investigated the effects of resveratrol on palmitate-induced inflammation and elucidated the underlying mechanisms in skeletal muscle cells. The results showed that palmitate significantly enhanced TNF-α and IL-6 mRNA expression and protein secretion from C2C12 cells at 12, 24, and 36\u2009h treatments. Increased expression of cytokines was accompanied by an enhanced phosphorylation of JNK, P38, ERK1/2, and IKKα/IKKβ. In addition, JNK and P38 inhibitors could significantly attenuate palmitate-induced mRNA expression of TNF-α and IL-6, respectively, whereas NF-κB inhibitor reduced the expression of both cytokines in palmitate-treated cells. Resveratrol pretreatment significantly prevented palmitate-induced TNF-α and IL-6 mRNA expression and protein secretion in C2C12 cells. Importantly, pre-treatment of the cells with resveratrol completely abrogated the phosphorylation of ERK1/2, JNK, and IKKα/IKKβ in palmitate treated cells. The protection from palmitate-induced inflammation by resveratrol was accompanied by a decrease in the generation of reactive species (ROS). N-acetyl cysteine (NAC), a known scavenger of ROS, could protect palmitate-induced expression of TNF-α and IL-6. Furthermore, inhibition of SIRT1 by shRNA or sirtinol demonstrated that the anti-inflammatory effect of resveratrol in muscle cells is mediated through a SIRT1-independent mechanism. Taken together, these findings suggest that resveratrol may represent a promising therapy for prevention of inflammation in skeletal muscle cells. J. Cell. Biochem. 118: 2654-2663, 2017.© 2017 Wiley Periodicals, Inc.

Keyword: oxygen

Effect of cross linking of PVA/starch and reinforcement of modified barley husk on the properties of composite films.

Barley husk (BH) was graft copolymerized by . The crystalline behavior of BH decreased after grafting. Poly vinyl alcohol (PVA)/starch (St) blend film, urea formaldehyde cross linked PVA/St films and composite films containing natural BH, grafted BH were prepared separately. The effect of urea/starch ratio, content of BH and grafted BH on the mechanical properties, water uptake (%), and biodegradability of the composite films was observed. With increase in urea: starch ratio from 0 to 0.5 in the blend, tensile strength of cross linked film increased by 40.23% compared to the PVA/St film. However, in grafted BH composite film, the tensile strength increased by 72.4% than PVA/St film. The degradation rate of natural BH composite film was faster than PVA/St film. Various films were characterized by SEM, FT-IR and thermal analysis.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Oleate protects macrophages from palmitate-induced apoptosis through the downregulation of CD36 expression.

In obese patients, free fatty acids ectopically accumulated in non-adipose tissues cause cell death. Saturated fatty acids are more deleterious to non-adipose cells, and supplementation with monounsaturated fatty acids has been proposed to rescue cells from saturated fatty -induced cytotoxicity; however, the mechanisms are not well understood. To understand the cytoprotective role of monounsaturated fatty acids in lipotoxic cell death of macrophages, we investigated the antagonizing effect of oleate and the underlying mechanisms in palmitate-treated RAW264.7\xa0cells. Palmitate strongly induced apoptosis in macrophages by increasing CD36 expression, which was identified to mediate both endoplasmic reticulum stress and the generation of reactive species. Co-treatment with oleate significantly reduced CD36 expression and its downstream signaling pathways of apoptosis in palmitate-treated cells. These findings provide a novel mechanism by which oleate protects macrophages from palmitate-induced lipotoxicity.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: oxygen

Inhibition of protein kinase R protects against -induced inflammation, oxidative stress, and apoptosis through the JNK/NF-kB/NLRP3 pathway in cultured H9C2 cardiomyocytes.

Double-stranded RNA-dependent protein kinase (PKR) is a critical regulator of apoptosis, oxidative stress, and inflammation under hyperlipidemic and insulin resistance conditions. Saturated free fatty acids, such as (PA), are known inducers of apoptosis in numerous cell types. However, the underlying molecular mechanism is not fully understood. The aim of the present study was to examine the effect of PA on cultured rat H9C2 cardiac myocytes cells and to investigate the PKR mediated harmful effects of PA in vitro in cultured cardiomyocytes.PKR expression was determined by immunofluorescence and immunoblotting. Oxidative stress and apoptosis were determined by flow cytometry and assay kits. The expression of different gene markers of apoptosis, oxidative stress, and inflammation were measured by Western blot analysis and reverse transcription polymerase chain reaction.PKR expression, reactive species levels as well as apoptosis were increased in PA-treated cultured H9C2 cardiomyocytes. The harmful effects of PA were attenuated by a selective PKR inhibitor, C16. Moreover, we observed that upregulation of c-Jun N-terminal kinase (JNK), nuclear factor-kB (NF-kB) and NACHT, LRR and PYD domains-containing protein 3 (NLRP3) pathways is associated with increased expression of interleukin 6 and tumor necrosis factor-α in PA-treated cardiomyocytes and attenuation by a selective PKR inhibitor.Our study reports, for the first time, that PKR-mediated harmful effects of PA in cultured cardiomyocytes via activation of JNK, NF-kB, and NLRP3 pathways. Inhibition of PKR is one of the possible mechanistic approaches to inhibit inflammation, oxidative stress, and apoptosis in lipotoxicity-induced cardiomyocyte damage.© 2018 Wiley Periodicals, Inc.

Keyword: oxygen

Protective Effects and Mechanism of Meretrix meretrix Oligopeptides against Nonalcoholic Fatty Liver Disease.

oligopeptides (MMO) derived from shellfish have important medicinal properties. We previously obtained MMO from alcalase by hydrolysis processes. Here we examine the protective effects of MMO against nonalcoholic fatty liver disease (NAFLD) and explored the underlying mechanism. Human Chang liver cells were used in our experiments after exposure to at a final concentration of 15 μg/mL for 48 h to induce an overload of fatty as NAFLD model cells. Treatment with MMO for 24 h increased the viability of the NAFLD model cells by inhibiting apoptosis. MMO alleviated oxidative stress in the NAFLD model cells by preserving reactive species activity and increasing malondialdehyde and superoxide dismutase activity. MMO improved mitochondrial dysfunction by decreasing the mitochondrial membrane potential and increasing the activities of Na⁺/K⁺-ATPase and Ca/Mg-ATPase. In addition, MMO inhibited the activation of cell death-related pathways, based on reduced p-JNK, Bax expression, tumor necrosis factor-α, caspase-9, and caspase-3 activity in the NAFLD model cells, and Bcl-2 expression was enhanced in the NAFLD model cells compared with the control group. These findings indicate that MMO have antioxidant and anti-apoptotic effects on NAFLD model cells and may thus exert protective effects against NAFLD.

Keyword: oxygen

Dihydromyricetin inhibits NLRP3 inflammasome-dependent pyroptosis by activating the Nrf2 signaling pathway in vascular endothelial cells.

Increasing evidence demonstrates that pyroptosis, pro-inflammatory programmed cell death, is linked to atherosclerosis; however, the underlying mechanisms remain to be elucidated. Dihydromyricetin (DHM), a natural flavonoid, was reported to exert anti-oxidative and anti-inflammatory bioactivities. However, the effect of DHM on atherosclerosis-related pyroptosis has not been studied. In the present study, (PA) treatment led to pyroptosis in human umbilical vein endothelial cells (HUVECs), as evidenced by caspase-1 activation, LDH release, and propidium iodide-positive staining; enhanced the maturation and release of proinflammatory cytokine IL-1β and activation of the NLRP3 inflammasome; and markedly increased intracellular reactive species (ROS) and mitochondrial ROS (mtROS) levels. Moreover, NLRP3 siRNA transfection or treatment with inhibitors efficiently suppressed PA-induced pyroptosis, and pretreatment with total ROS scavenger or mtROS scavenger attenuated PA-induced NLRP3 inflammasome activation and subsequent pyroptosis. However, DHM pretreatment inhibited PA-induced pyroptotic cell death by increasing cell viability, decreasing LDH and IL-1β release, improving cell membrane integrity, and abolishing caspase-1 cleavage and subsequent IL-1β maturation. We also found that DHM pre-treatment remarkably reduced the levels of intracellular ROS and mtROS and activated the Nrf2 signaling pathway. Moreover, knockdown of Nrf2 by siRNA abrogated the inhibitory effects of DHM on ROS generation and subsequent PA-induced pyroptosis. Together, these results indicate that the Nrf2 signaling pathway plays a role, as least in part, in the DHM-mediated improvement in PA-induced pyroptosis in vascular endothelial cells, which implies the underlying medicinal value of DHM targeting immune/inflammatory-related diseases, such as atherosclerosis.© 2017 BioFactors, 44(2):123-136, 2018.

Keyword: oxygen

The use of stable isotope ratio analysis to characterise saw palmetto (Serenoa Repens) extract.

Saw palmetto extract (SPE) has many pharmacological effects. Thus, its demand and value has increased steadily, along with the presence of counterfeit SPEs on the market. In this work bulk δC, δH, δO and fatty δC, δH analysis was performed in 20 authentic and 9 commercial SPEs, 12 meat fats and 4 pure fatty acids. Authentic SPEs are characterised by bulk values from -31.0‰ to -29.7‰ for δC, -176‰ to -165‰ for δH, 27.2‰ to 40.7‰ for δO, and values of capric, caprylic, lauric, myristic, and oleic acids from -37.4‰ to -30.5‰ for δC and -187‰ to -136‰ for δH. The isotopic values of all the commercial SPEs were out of these ranges and more similar to those of meat fat and pure fatty acids. Stable isotope ratio analysis can therefore be proposed as a suitable tool for detecting adulteration in SPEs.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Uncoupling effect of palmitate is exacerbated in skeletal muscle mitochondria of sea-acclimatized king penguins (Aptenodytes patagonicus).

In king penguin juveniles, the environmental transition from a terrestrial to a marine habitat, occurring at fledging, drastically stimulates lipid catabolism and the remodelling of muscle mitochondria to sustain extensive swimming activity and thermoregulation in the cold circumpolar oceans. However, the exact nature of these mechanisms remains only partially resolved. Here we investigated, in vitro, the uncoupling effect of increasing doses of fatty acids in pectoralis muscle intermyofibrillar mitochondria isolated, either from terrestrial never-immersed or experimentally cold water immersed pre-fledging king penguins or from sea-acclimatized fledged penguins. Mitochondria exhibited much greater palmitate-induced uncoupling respiration and higher maximal oxidative capacity after acclimatization to marine life. Such effects were not reproduced experimentally after repeated immersions in cold water, suggesting that the plasticity of mitochondrial characteristics may not be primarily driven by cold exposure per se but by other aspects of sea acclimatization.Copyright © 2017. Published by Elsevier Inc.

Keyword: oxygen

Peroxisome proliferator-activated receptor-delta agonist ameliorated inflammasome activation in nonalcoholic fatty liver disease.

To evaluate the inflammasome activation and the effect of peroxisome proliferator-activated receptors (PPAR)-δ agonist treatment in nonalcoholic fatty liver disease (NAFLD) models.Male C57BL/6J mice were classified according to control or high fat diet (HFD) with or without PPAR-δ agonist (GW) over period of 12 wk [control, HFD, HFD + lipopolysaccharide (LPS), HFD + LPS + GW group]. HepG2 cells were exposed to (PA) and/or LPS in the absence or presence of GW.HFD caused glucose intolerance and hepatic steatosis. In mice fed an HFD with LPS, caspase-1 and interleukin (IL)-1β in the liver were significantly increased. Treatment with GW ameliorated the steatosis and inhibited overexpression of pro-inflammatory cytokines. In HepG2 cells, PA and LPS treatment markedly increased mRNA of several nucleotide-binding and oligomerization domain-like receptor family members (NLRP3, NLRP6, and NLRP10), caspase-1 and IL-1β. PA and LPS also exaggerated reactive species production. All of the above effects of PA and LPS were reduced by GW. GW also enhanced the phosphorylation of AMPK-α.PPAR-δ agonist reduces fatty -induced inflammation and steatosis by suppressing inflammasome activation. Targeting the inflammasome by the PPAR-δ agonist may have therapeutic implication for NAFLD.

Keyword: oxygen

Integrating Extracellular Flux Measurements and Genome-Scale Modeling Reveals Differences between Brown and White Adipocytes.

White adipocytes are specialized for energy storage, whereas brown adipocytes are specialized for energy expenditure. Explicating this difference can help identify therapeutic targets for obesity. A common tool to assess metabolic differences between such cells is the Seahorse Extracellular Flux (XF) Analyzer, which measures consumption and media acidification in the presence of different substrates and perturbagens. Here, we integrate the Analyzer\'s metabolic profile from human white and brown adipocytes with a genome-scale metabolic model to predict flux differences across the metabolic map. Predictions matched experimental data for the metabolite 4-aminobutyrate, the protein ABAT, and the fluxes for glucose, glutamine, and palmitate. We also uncovered a difference in how adipocytes dispose of nitrogenous waste, with brown adipocytes secreting less ammonia and more urea than white adipocytes. Thus, the method and software we developed allow for broader metabolic phenotyping and provide a distinct approach to uncovering metabolic differences.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: oxygen

Effect of metabolic stressors on survival and growth of in\xa0vitro cultured ovine preantral follicles and enclosed oocytes.

The present study was undertaken to study the effect of metabolic stressors like elevated levels of ammonia, urea, Non-esterified fatty (NEFA) and β-hydroxybutyric (BHB) on preantral follicle growth, survival, growth rates of oocytes enclosed in preantral follicles (PFs), maturation rates of oocytes recovered from cultured follicles, hormone production (estrogen and progesterone), reactive species (ROS) as well as superoxide dismutase (SOD) activity. Small pre-antral follicles (SPFs, 100-250\xa0μm) and large pre-antral follicles (LPFs, 250-450\xa0μm) were isolated from slaughterhouse ovaries by a mechanical cum enzymatic method. SPFs and LPFs were cultured in\xa0vitro for 14 and 7 days respectively and examined for their growth, survival and growth rates of enclosed oocytes in PFs exposed with different concentration of ammonia (0, 100, 150, 200, 250, 300 and 400\xa0μM), urea (0, 4, 4.5, 5, 5.5,6, 7 and 8\xa0mM), NEFA [Basal NEFA (70\xa0μM): stearic , SA (10\xa0μM)+, PA(20\xa0μM)+oleic , OA(40\xa0μM), b) Medium combo (140\xa0μM): SA (20\xa0μM)+ PA(40\xa0μM)+ OA(80\xa0μM), c) High combo (210\xa0μM): SA (30\xa0μM)+PA(60\xa0μM)+OA(120\xa0μM), d) Very high Combo (280\xa0μM): SA(40\xa0μM)+PA(80\xa0μM)+OA(160\xa0μM)] and BHB (0, 0.5, 0.75, and 1\xa0μM). Results indicated that ammonia, urea, NEFA and BHB caused inhibition of survival and growth of in\xa0vitro cultured ovine PFs and enclosed oocytes at the levels of 300\xa0μM, 8\xa0mM, high combo level of NEFA and 0.75\xa0μM respectively. Our study may contribute to the identification of the mechanisms involved in decline of fertility due to metabolic and nutritional stress in ruminants.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: oxygen

Phycocyanin-Functionalized Selenium Nanoparticles Reverse -Induced Pancreatic β Cell Apoptosis by Enhancing Cellular Uptake and Blocking Reactive Species (ROS)-Mediated Mitochondria Dysfunction.

Accumulation of (PA) in human bodies could cause damage to pancreatic β cells and lead to chronic diseases by generation of reactive species (ROS). Therefore, it is of great significance to search for nutrition-available agents with antioxidant activity to protect pancreatic islet cells against PA-induced damage. Phycocyanin (PC) and selenium (Se) have been reported to have excellent antioxidant activity. In this study, PC-functionalized selenium nanoparticles (PC-SeNPs) were synthesized to investigate the in vitro protective effects on INS-1E rat insulinoma β cells against PA-induced cell death. A potent protective effect was achieved by regulation of particle size and PC content. Among three PC-SeNPs (165, 235, and 371 nm), PC-SeNPs-235 nm showed the highest cellular uptake and the best protective activities. For cell cycle analysis, PC-SeNPs showed a better protective effect on PA-induced INS-1E cell apoptosis than PC or SeNPs, and PC-SeNPs-235 nm exhibited the best effect. Further mechanistic studies demonstrated that PA induced overproduction of intracellular ROS, mitochondria fragmentation, activation of caspase-3, -8, and -9, and cleavage of PARP. However, pretreatment of the cells with PC-SeNPs effectively blocked these intracellular events, which suggests that PC-SeNPs could protect INS-1E cells against PA-induced cell apoptosis via attenuating oxidative stress and downstream signaling pathways. This finding provides a great promising nutritional approach for protection against diseases related to islet damage.

Keyword: oxygen

Oleic/Palmitate Induces Apoptosis in Human Articular Chondrocytes via Upregulation of NOX4 Expression and ROS Production.

The association between obesity and reactive species (ROS) production in osteoarthritis (OA) patients has already been identified. However, the specific mechanism underlying ROS production and OA progression has never been elucidated. Osteoarthritic cartilage was obtained from patients undergoing total hip arthroplasty, and chondrocytes were isolated from these tissues. The cells were treated with varying concentrations (10,100,500 μM, and 5 mM) of oleic/palmitate (O/P) mixture at different times, that is at 6, 24, and 48 h. Cell viability was determined using MTT assay. ROS production was detected using immunofluorescence and flow cytometry. The protein levels of NOX4 and cleaved-caspase3 were detected using Western blot assay. O/P significantly decreased cell viability at 10, 100, 500 μM, and 5 mM in a dose-dependent manner. Furthermore, the cell viability was reduced by 500 μM O/P mixture at 6, 24, and 48 h in a time-dependent manner. Pretreatment with 500 μM O/P significantly enhanced ROS production and cell apoptosis in chondrocytes. Furthermore, treatment with O/P mixture significantly enhanced the expression of NOX4 and caspase3 activation in a dose- and time- dependent manner. More importantly, inhibition of NOX4 could partially eliminate O/P-induced chondrocytes apoptosis by reducing ROS production. To conclude, O/P treatment enhances ROS production and cell apoptosis mainly by upregulating the protein levels of NOX4 and caspase3 activation in chondrocytes, indicating a potential therapeutic target of OA in obesity patients.© 2016 by the Association of Clinical Scientists, Inc.

Keyword: oxygen

Resveratrol Maintains Lipid Metabolism Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver lipid metabolism disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver lipid metabolism disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid metabolic disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular lipid metabolic imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of lipid metabolic disorders relevant to the circadian clock.

Keyword: oxygen

Hepatic stimulator substance inhibits calcium overflow through the mitochondria-associated membrane compartment during nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease is considered a disorder of the endoplasmic reticulum (ER) and mitochondria. Recent studies have shown that the ER and mitochondrial membranes overlap by 15-20%, a region referred to as the \'mitochondria-associated ER membrane\' (MAM). Some proteins, including sarco/ER calcium ATPase (SERCA), are located in the MAM and have an important role in Ca signaling and homeostasis between the ER and the mitochondria. Our previous study showed that hepatic stimulator substance (HSS) inhibits the ER stress induced by reactive species, thus reducing mitochondrial damage. However, the mechanism underlying the protective effect of HSS on the ER and ER-mitochondrial interaction remains unclear. In this study, we confirmed that the exogenous expression of HSS protected the liver from steatosis in mice with nonalcoholic steatohepatitis. More importantly, the protection provided by HSS allowed SERCA in the MAM compartment to function well, preventing the extensive influx of cytosolic free Ca to the mitochondria, thus preserving the mitochondrial functions from calcium overload and relieving -induced hepatocyte steatosis. Our results suggest that the protective effect of HSS on SERCA expression is associated with the maintenance of calcium homeostasis within the MAM, thus ameliorating the disordered Ca communication between the ER and mitochondria.

Keyword: oxygen

Activation of Nrf2 by Phloretin Attenuates -Induced Endothelial Cell Oxidative Stress via AMPK-Dependent Signaling.

Phloretin, a dihydrochalcone structural flavonoid compound, possesses antioxidant activity. In this study, we conducted studies to explore the function of phloretin on high -induced oxidative stress in human umbilical vein endothelial cells and investigated the potential mechanism using ribonucleic sequencing (RNA-Seq). Our findings reveal that phloretin significantly decreased the levels of intracellular reactive species (ROS) and malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione peroxidase-1 (Gpx-1) activity, and restored the loss of mitochondrial membrane potential (MMP). Next, whole transcriptome analysis was performed using RNA-Seq The results indicated more than 3000 differentially expressed genes (DEGs). Gene Ontology analysis revealed that the DEGs were categorized functionally, mainly by the biological processes, cell metabolism, and cellular response to chemical stimulus. The Kyoto Encyclopedia of Genes and Genomes indicated that they were mainly enriched in cAMP, apoptosis, and cytoskeletal regulation signaling pathways. Furthermore, on the basis of the results of RNA-Seq and Western blotting, our study verified that phloretin upregulated the expression of p-Nrf2 and HO-1 by promoting the phosphorylation of AMPK at Thr through activation of liver kinase B1. In conclusion, phloretin attenuates PA-induced oxidative stress in HUVECs via the AMPK/Nrf2 antioxidative pathway.

Keyword: oxygen

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy metabolism, but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: oxygen

Serine prevented high-fat diet-induced oxidative stress by activating AMPK and epigenetically modulating the expression of glutathione synthesis-related genes.

Serine deficiency has been observed in patients with nonalcoholic fatty liver disease (NAFLD). Whether serine supplementation has any beneficial effects on the prevention of NAFLD remains unknown. The present study was conducted to investigate the effects of serine supplementation on hepatic oxidative stress and steatosis and its related mechanisms. Forty male C57BL/6J mice (9week-old) were randomly assigned into four groups (n=10) and fed: i) a low-fat diet; ii) a low-fat diet supplemented with 1% (wt:vol) serine; iii) a high-fat (HF) diet; and iv) a HF diet supplemented with 1% serine, respectively. (PA)-treated primary hepatocytes separated from adult mice were also used to study the effects of serine on oxidative stress. The results showed that serine supplementation increased glucose tolerance and insulin sensitivity, and protected mice from hepatic lipid accumulation, but did not significantly decreased HF diet-induced weight gain. In addition, serine supplementation protected glutathione (GSH) antioxidant system and prevented hypermethylation in the promoters of glutathione synthesis-related genes, while decreasing reactive species (ROS) in mice fed a HF diet. Moreover, we found that serine supplementation increased phosphorylation and S-glutathionylation of AMP-activated protein kinase α subunit (AMPKα), and decreased ROS, malondialdehyde and triglyceride contents in PA-treated primary hepatocytes. However, while AMPK activity or GSH synthesis was inhibited, the abovementioned effects of serine on PA-treated primary hepatocytes were not observed. Our results suggest that serine supplementation could prevent HF diet-induced oxidative stress and steatosis by epigenetically modulating the expression of glutathione synthesis-related genes and through AMPK activation.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: oxygen

Nerve growth factor protects against -induced injury in retinal ganglion cells.

Accumulating evidence supports an important role for nerve growth factor (NGF) in diabetic retinopathy. We hypothesized that NGF has a protective effect on rat retinal ganglion RGC-5 cells injured by (PA), a metabolic factor implicated in the development of diabetes and its complications. Our results show that PA exposure caused apoptosis of RGC-5 cells, while NGF protected against PA insult in a concentration-dependent manner. Additionally, NGF significantly attenuated the levels of reactive species (ROS) and malondialdehyde (MDA) in RGC-5 cells. Pathway inhibitor tests showed that the protective effect of NGF was completely reversed by LY294002 (PI3K inhibitor), Akt VIII inhibitor, and PD98059 (ERK1/2 inhibitor). Western blot analysis revealed that NGF induced the phosphorylation of Akt/FoxO1 and ERK1/2 and reversed the PA-evoked reduction in the levels of these proteins. These results indicate that NGF protects RGC-5 cells against PA-induced injury through anti-oxidation and inhibition of apoptosis by modulation of the PI3K/Akt and ERK1/2 signaling pathways.

Keyword: oxygen

Resveratrol attenuates oxidative injury in human umbilical vein endothelial cells through regulating mitochondrial fusion via TyrRS-PARP1 pathway.

Oxidative stress-induced damage in endothelial cells is a crucial initiator of atherosclerosis (AS), which is highly related to excessive reactive species (ROS) and mitochondrial dynamics. Resveratrol (RSV) exerts beneficial effects against endothelial oxidative injury, while the underlying mechanisms have not been fully elucidated. Thus, we aimed to explore the role of mitochondria dynamics during the anti-oxidative activities of RSV in (PA)-stimulated human umbilical vein endothelial cells (HUVECs) and to verify whether tyrosyl transfer- RNA synthetase (TyrRS) and poly (ADP-ribose) polymerase 1 (PARP1) are targeted during this process.HUVECs were exposed to 200\u2009μM of PA for 16\u2009h before treated with 10\u2009μM of RSV for 8\u2009h. Cell viability was detected using Cell counting kit-8 (CCK-8) assay. The intracellular ROS level and mitochondria membrane potential (MMP) were measured using microplate reader and flow cytometry. The malondialdehyde and superoxide dismutase were measured using the microplate reader. The mitochondrial morphology and fusion process was observed under transmission electron microscopy and confocal microscopy. TyrRS and PARP1 were knocked down with the specific small interference RNAs (siRNA), and the protein expressions of TyrRS, PARP1, and mitochondrial fusion proteins (MFN1, MFN2, and OPA1) were measured by western blot.RSV treatment suppressed the PA-induced injuries in HUVECs, including the damage to cell viability, oxidative stress, and loss of MMP. Additionally, RSV improved the protein levels of MFN1, MFN2, and OPA1 as well as inhibited the PA-induced fragmentation of mitochondria. However, the effects of RSV on oxidative stress and mitochondrial fusion were abolished by the pretreatment of siRNAs of TyrRS and PARP1, indicating that these effects of RSV were dependent on the TyrRS-PARP1 pathway.RSV attenuated endothelial oxidative injury by regulating mitochondrial fusion via TyrRS-PARP1 signaling pathway.

Keyword: oxygen

Oleic stimulation of motility of human extravillous trophoblast cells is mediated by stearoyl-CoA desaturase-1 activity.

Do fatty acids regulate development and motility of human extravillous trophoblast cells (EVTs)?Oleic is a promising lipid molecule that has beneficial effects on motility and development of human EVTs.Fatty uptake into trophoblast cells is important for maintaining cellular events during pregnancy, but the molecular mechanisms of action of various fatty acids, including trans fatty acids, saturated fatty acids and monounsaturated fatty acids, in EVT cell lines are not clear.Effects of oleic , elaidic , and stearic on HTR8/SVneo cells were assessed in diverse assays in a dose- and time-dependent manner.Effects of fatty acids on cell proliferation, migration, invasion and apoptosis (Annexin V expression, propidium iodide staining, TUNEL and invasion assays) of HTR8/SVneo cells were determined. Signal transduction pathways in HTR8/SVneo cells in response to fatty acids were determined by Western blot analyses. Regulation of fatty acids on oxidative conditions in EVTs were determined and validated by measurement of production of cellular reactive species, intracellular concentrations of free Ca2+and lipid peroxidation assays.In present study, we confirmed different effects of oleic and elaidic on migration, invasion, proliferation and apoptosis of the EVT cell line, HTR8/SVneo. We also investigated stearoyl-CoA desaturase-1 (SCD1) to determine if its activity contributed to oleic -induced migration of HTR8/SVneo cells. Next, we analyzed cell signaling molecules mediated by oleic and elaidic treatment, including MAPK and PI3K/AKT pathways in HTR8/SVneo cells. We further established whether selective inhibition of signaling molecules altered the ability of fatty acids to cause changes in migration and proliferation of HTR8/SVneo cells. Last, we examined the regulatory effects of oleic and SCD1 on oxidative stress in HTR8/SVneo cells.N/A.The lack of in vivo animal studies is a major limitation of this research. Effectiveness of oleic to stimulate migration of human EVT cells requires further investigation.Our results suggest that oleic can play an important role in promoting invasion of human EVT cell lines while both trans fatty acids and saturated fatty acids are not conducive to normal placentation. This may have implications for the prevention of pre-eclampsia and intrauterine growth restriction.This work was supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (No. HI15C0810) awarded to G.S. and (No. HI17C0929) awarded to W.L. There are no conflicts of interest.© The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Keyword: oxygen

Graphene oxide nano-bio interaction induces inhibition of spermatogenesis and disturbance of fatty metabolism in the nematode Caenorhabditis elegans.

Graphene oxide (GO) has the potential for wide applications, which necessitates an intensive investigation of its potential hazard on human and environmental health. Even if previous studies show reproductive toxicity in the nematode Caenorhabditis elegans, the mechanisms of reproductive toxicity by GO are poorly understood. To understand the underlying mechanisms of GO-induced reproductive toxicity, we investigated the interaction between GO and C. elegans using Raman spectroscopy, sperm counts produced by spermatogenesis, progeny and analyzed the fatty metabolism using molecular techniques. GO-characteristic Raman spectral bands measured throughout C. elegans, brood size and Hoecst staining of dissected gonads clearly showed GO accumulation in the reproductive organs, reduced progeny and low sperm counts, which are possibly direct results of the reproductive toxicity from GO exposure. Interestingly, reduced fatty metabolites, such as stearic, oleic, palmitoleic, and acids, were found with GO exposure. We found that GO increased intestinal fat accumulation in wild type N2, fat-5(tm420), and fat-7(wa36) mutants, whereas it decreased fat storage in the fat-6(tm331) and nhr-49(nr2041) mutants. GO exposure affected C. elegans fat accumulation and consumption, which was possibly regulated by daf-16 and nhr-80 gene activity. Also, GO exposure suppressed the survival of long-lived fat-5(tm420) mutants, whereas it increased the survival of short-lived nhr-49(nr2041) mutants. Hence, our studies collectively indicated that GO accumulation in reproductive organs, suppression of spermatogenesis, and the alteration of fatty metabolism play critical roles in understanding mechanisms of toxicity in C. elegans.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: oxygen

Free fatty receptor 1 (FFAR1/GPR40) signaling affects insulin secretion by enhancing mitochondrial respiration during palmitate exposure.

Fatty acids affect insulin secretion via metabolism and FFAR1-mediated signaling. Recent reports indicate that these two pathways act synergistically. Still it remains unclear how they interrelate. Taking into account the key role of mitochondria in insulin secretion, we attempted to dissect the metabolic and FFAR1-mediated effects of fatty acids on mitochondrial function. One-hour culture of MIN6 cells with palmitate significantly enhanced mitochondrial respiration. Antagonism or silencing of FFAR1 prevented the palmitate-induced rise in respiration. On the other hand, in the absence of extracellular palmitate FFAR1 agonists caused a modest increase in respiration. Using an agonist of the M3 muscarinic acetylcholine receptor and PKC inhibitor we found that in the presence of the fatty mitochondrial respiration is regulated via Gαq protein-coupled receptor signaling. The increase in respiration in palmitate-treated cells was largely due to increased glucose utilization and oxidation. However, glucose utilization was not dependent on FFAR1 signaling. Collectively, these results indicate that mitochondrial respiration in palmitate-treated cells is enhanced via combined action of intracellular metabolism of the fatty and the Gαq-coupled FFAR1 signaling. Long-term palmitate exposure reduced ATP-coupling efficiency of mitochondria and deteriorated insulin secretion. The presence of the FFAR1 antagonist during culture did not improve ATP-coupling efficiency, however, it resulted in enhanced mitochondrial respiration and improved insulin secretion after culture. Taken together, our study demonstrates that during palmitate exposure, integrated actions of fatty metabolism and fatty -induced FFAR1 signaling on mitochondrial respiration underlie the synergistic action of the two pathways on insulin secretion.Copyright © 2015. Published by Elsevier B.V.

Keyword: oxygen

Glucose fluctuation increased hepatocyte apoptosis under lipotoxicity and the involvement of mitochondrial permeability transition opening.

Oxidative stress is considered to be an important factor in producing lethal hepatocyte injury associated with nonalcoholic fatty liver disease (NAFLD). Glucose fluctuation, more pronounced in patients with diabetes, has been recognized as an even stronger oxidative stress inducer than the sustained hyperglycemia. Here, we investigated the role of glucose variability in the development of the NAFLD based on hepatocyte apoptosis and possible mechanisms. To achieve this goal we studied C57BL/6J mice that were maintained on a high fat diet (HFD) and injected with glucose (3 g/kg) twice daily to induce intermittent high glucose (IHG). We also studied hepatic L02 cells incubated with (PA) to induce steatosis. The following experimental groups were compared: normal glucose (NG), sustained high glucose (SHG) and IHG with or without PA. We found that, although hepatic enzyme levels and liver lipid deposition were comparable between HFD mice injected with glucose or saline, the glucose injected mice displayed marked hepatocyte apoptosis and inflammation, accompanied by increased lipid peroxide in liver. In vitro, in the presence of PA, IHG increased L02 cell apoptosis and oxidative stress and produced pronounced mitochondrial dysfunction relative to the NG and SHG groups. Furthermore, treatment with the mitochondrial permeability transition (MPT) inhibitor, cyclosporin A (1.5 μmol/l), prevented mitochondrial dysfunction, oxidative stress and hepatocyte apoptosis. Our data suggests that IHG under lipotoxicity might contribute to the development of NAFLD by increasing oxidative stress and hepatocyte apoptosis via MPT and its related mitochondrial dysfunction.© 2015 Society for Endocrinology.

Keyword: oxygen

Protein Analysis of Sapienic -Treated Porphyromonas gingivalis Suggests Differential Regulation of Multiple Metabolic Pathways.

Lipids endogenous to skin and mucosal surfaces exhibit potent antimicrobial activity against Porphyromonas gingivalis, an important colonizer of the oral cavity implicated in periodontitis. Our previous work demonstrated the antimicrobial activity of the fatty sapienic (C(16:1Δ6)) against P. gingivalis and found that sapienic treatment alters both protein and lipid composition from those in controls. In this study, we further examined whole-cell protein differences between sapienic -treated bacteria and untreated controls, and we utilized open-source functional association and annotation programs to explore potential mechanisms for the antimicrobial activity of sapienic . Our analyses indicated that sapienic treatment induces a unique stress response in P. gingivalis resulting in differential expression of proteins involved in a variety of metabolic pathways. This network of differentially regulated proteins was enriched in protein-protein interactions (P = 2.98 × 10(-8)), including six KEGG pathways (P value ranges, 2.30 × 10(-5) to 0.05) and four Gene Ontology (GO) molecular functions (P value ranges, 0.02 to 0.04), with multiple suggestive enriched relationships in KEGG pathways and GO molecular functions. Upregulated metabolic pathways suggest increases in energy production, lipid metabolism, iron acquisition and processing, and respiration. Combined with a suggested preferential metabolism of serine, which is necessary for fatty biosynthesis, these data support our previous findings that the site of sapienic antimicrobial activity is likely at the bacterial membrane.P. gingivalis is an important opportunistic pathogen implicated in periodontitis. Affecting nearly 50% of the population, periodontitis is treatable, but the resulting damage is irreversible and eventually progresses to tooth loss. There is a great need for natural products that can be used to treat and/or prevent the overgrowth of periodontal pathogens and increase oral health. Sapienic is endogenous to the oral cavity and is a potent antimicrobial agent, suggesting a potential therapeutic or prophylactic use for this fatty . This study examines the effects of sapienic treatment on P. gingivalis and highlights the membrane as the likely site of antimicrobial activity.Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Keyword: oxygen

Effects of Fatty Acids on Intracellular [Ca2+], Mitochondrial Uncoupling and Apoptosis in Rat Pachytene Spermatocytes and Round Spermatids.

The aim of this work was to explore the ability of free arachidonic , and the unsaturated fatty acids oleic and docosahexaenoic to modify calcium homeostasis and mitochondrial function in rat pachytene spermatocytes and round spermatids. In contrast to , unsaturated fatty acids produced significant increases in intracellular calcium concentrations ([Ca2+]i) in both cell types. Increases were fatty specific, dose-dependent and different for each cell type. The arachidonic effects on [Ca2+]i were higher in spermatids than in spermatocytes and persisted when residual extracellular Ca2+ was chelated by EGTA, indicating that the increase in [Ca2+]i originated from release of intracellular calcium stores. At the concentrations required for these increases, unsaturated fatty acids produced no significant changes in the plasma membrane potential of or non-specific permeability in spermatogenic cells. For the case of arachidonic , the [Ca2+]i increases were not caused by its metabolic conversion to eicosanoids or anandamide; thus we attribute this effect to the fatty itself. As estimated with fluorescent probes, unsaturated fatty acids did not affect the intracellular pH but were able to induce a progressive decrease in the mitochondrial membrane potential. The association of this decrease with reduced reactive species (ROS) production strongly suggests that unsaturated fatty acids induced mitochondrial uncoupling. This effect was stronger in spermatids than in spermatocytes. As a late event, arachidonic induced caspase 3 activation in a dose-dependent manner both in the absence and presence of external Ca2+. The concurrent but differential effects of unsaturated fatty acids on [Ca2+]i and mitochondrial functions are additional manifestations of the metabolic changes that germ cells undergo during their differentiation.

Keyword: oxygen

Characterisation and attempted differentiation of European and extra-European olive oils using stable isotope ratio analysis.

European law requires a designation of origin for virgin and extra virgin olive oils (at least in terms of EU/non-EU provenance). Stable isotope ratios have been successfully applied to determine the geographical origin of olive oils, but never to distinguish EU and non-EU oils. In this study H/H, C/C and O/O ratios were analysed in bulk olive oils using Isotope Ratio Mass Spectrometry (IRMS) as well as C/C and H/H in the four main fatty acids (linoleic, oleic, and stearic acids) using IRMS coupled with GC. The isotopic composition of olive oils was successfully used to distinguish samples originating in the two areas. Specifically, when bulk data were combined with fatty isotopic data the differentiation power of the method improved clearly. This separation is due to the specific isotopic fingerprint of the individual countries making up the EU and non-EU samples.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Negligible effect of eNOS palmitoylation on fatty regulation of contraction in ventricular myocytes from healthy and hypertensive rats.

S-palmitoylation is an important post-translational modification that affects the translocation and the activity of target proteins in a variety of cell types including cardiomyocytes. Since endothelial nitric oxide synthase (eNOS) is known to be palmitoylated and the activity of eNOS is essential in fatty -dependent β-oxidation in muscle, we aimed to test whether palmitoylation of eNOS is involved in (PA) regulation of left ventricular (LV) myocyte contraction from healthy (sham) and hypertensive (HTN) rats. Our results showed that PA, a predominant metabolic substrate for cardiac β-oxidation, significantly increased contraction and consumption rate (OCR) in LV myocytes from sham. Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) or eNOS gene deletion prevented PA regulation of the myocyte contraction or OCR, indicating the pivotal role of eNOS in mediating the effects of PA in cardiac myocytes. PA increased the palmitoylation of eNOS in LV myocytes and depalmitoylation with 2-bromopalmitate (2BP; 100\xa0μM) abolished the increment. Furthermore, although PA did not increase eNOS-Ser, 2BP reduced eNOS-Ser with and without PA. Intriguingly, PA-induced increases in contraction and OCR were unaffected by 2BP treatment. In HTN, PA did not affect eNOS palmitoylation, eNOS-Ser, or myocyte contraction. However, 2BP diminished eNOS palmitoylation and eNOS-Ser in the presence and absence of PA but did not change myocyte contraction. Collectively, our results confirm eNOS palmitoylation in LV myocytes from sham and HTN rats and its upregulation by PA in sham. However, such post-transcriptional modification plays negligible role in PA regulation of myocyte contraction and mitochondrial activity in sham and HTN.

Keyword: oxygen

Autophagy Protects against -Induced Apoptosis in Podocytes in vitro.

Autophagy is a highly conserved degradation process that is involved in the clearance of proteins and damaged organelles to maintain intracellular homeostasis and cell integrity. Type 2 diabetes is often accompanied by dyslipidemia with elevated levels of free fatty acids (FFAs). Podocytes, as an important component of the filtration barrier, are susceptible to lipid disorders. The loss of podocytes causes proteinuria, which is involved in the pathogenesis of diabetic nephropathy. In the present study, we demonstrated that (PA) promoted autophagy in podocytes. We further found that PA increased the production of reactive species (ROS) in podocytes and that NAC (N-acetyl-cysteine), a potent antioxidant, significantly eliminated the excessive ROS and suppressed autophagy, indicating that the increased generation of ROS was associated with the -induced autophagy in podocytes. Moreover, we also found that PA stimulation decreased the mitochondrial membrane potential in podocytes and induced podocyte apoptosis, while the inhibition of autophagy by chloroquine (CQ) enhanced -induced apoptosis accompanied by increased ROS generation, and the stimulation of autophagy by rapamycin (Rap) remarkably suppressed -induced ROS generation and apoptosis. Taken together, these in vitro findings suggest that PA-induced autophagy in podocytes is mediated by ROS production and that autophagy plays a protective role against PA-induced podocyte apoptosis.

Keyword: oxygen

Identification of a fatty binding protein4-UCP2 axis regulating microglial mediated neuroinflammation.

Hypothalamic inflammation contributes to metabolic dysregulation and the onset of obesity. Dietary saturated fats activate microglia via a nuclear factor-kappa B (NFκB) mediated pathway to release pro-inflammatory cytokines resulting in dysfunction or death of surrounding neurons. Fatty binding proteins (FABPs) are lipid chaperones regulating metabolic and inflammatory pathways in response to fatty acids. Loss of FABP4 in peripheral macrophages via either molecular or pharmacologic mechanisms results in reduced obesity-induced inflammation via a UCP2-redox based mechanism. Despite the widespread appreciation for the role of FABP4 in mediating peripheral inflammation, the expression of FABP4 and a potential FABP4-UCP2 axis regulating microglial inflammatory capacity is largely uncharacterized. To that end, we hypothesized that microglial cells express FABP4 and that inhibition would upregulate UCP2 and attenuate (PA)-induced pro-inflammatory response. Gene expression confirmed expression of FABP4 in brain tissue lysate from C57Bl/6J mice and BV2 microglia. Treatment of microglial cells with an FABP inhibitor (HTS01037) increased expression of Ucp2 and arginase in the presence or absence of PA. Moreover, cells exposed to HTS01037 exhibited attenuated expression of inducible nitric oxide synthase (iNOS) compared to PA alone indicating reduced NFκB signaling. Hypothalamic tissue from mice lacking FABP4 exhibit increased UCP2 expression and reduced iNOS, tumor necrosis factor-alpha (TNF-α), and ionized calcium-binding adapter molecule 1 (Iba1; microglial activation marker) expression compared to wild type mice. Further, this effect is negated in microglia lacking UCP2, indicating the FABP4-UCP2 axis is pivotal in obesity induced neuroinflammation. To our knowledge, this is the first report demonstrating a FABP4-UCP2 axis with the potential to modulate the microglial inflammatory response.Copyright © 2017. Published by Elsevier Inc.

Keyword: oxygen

The effect of oleic and on induction of steatosis and cytotoxicity on rat hepatocytes in primary culture.

In vitro models serve as a tool for studies of steatosis. and oleic acids can induce steatosis in cultured hepatocytes. The aim of our study was to verify steatogenic and cytotoxic effects of (PA), oleic (OA) and their combinations as well as their impact on functional capacity of rat primary hepatocytes. Hepatocytes were exposed to OA or PA (0.125-2 mmol/l) or their combination at ratios of 3:1, 2:1 or 1:1 at the final concentrations of 0.5-1 mmol/l. Both OA and PA caused a dose-dependent increase in triacylglycerol content in hepatocytes. PA was more steatogenic at 0.25 and 0.5 mmol/l while OA at 0.75 and 1 mmol/l. PA exhibited a dose-dependent cytotoxic effect associated with ROS production, present markers of apoptosis and necrosis and a decrease in albumin production. OA induced a damage of the cytoplasmic membrane from 1 mM concentration. Mixture of OA and PA induced lower cytotoxicity with less weakened functional capacity than did PA alone. Extent of steatosis was comparable to that after exposure to OA alone. In conclusion, OA or combination of OA with PA is more suitable for simulation of simple steatosis than PA alone.

Keyword: oxygen

Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and .

Danzhi jiangtang capsule (DJC) is mainly used to treat type 2 diabetes mellitus and vascular complication. Our aim was to investigate whether the protective effects of DJC by oral administration on high-fat diet (HFD) and -induced damages of endothelial cells are related to oxidative stress and endoplasmic reticulum (ER) stress. Male Sprague-Dawley rats were randomly divided into standard chow diet (SCD), HFD, HFD plus DJC-low dose (HFD\u2009+\u2009DJCL) and HFD plus DJC-high dose treatment groups (HFD\u2009+\u2009DJCH). The latter three groups were given HFD feeding for three months, then the HFD\u2009+\u2009DJCL and HFD\u2009+\u2009DJCH rats were treated with DJC (500 and 1000\u2009mg/kg/day) via gavage for another two months respectively. Endothelium-dependent relaxations induced by acetylcholine were observed in isolated rat thoracic aortic ring preparation. Malondialdehyde (MDA), total-antioxidant capacity (T-AOC), superoxide dismutase (SOD), interleukin 1β (IL-1β), tumour necrosis factor α (TNFα), free fatty acids (FFA), endothelin-1 (ET-1) and nitric oxide (NO) levels in serum were assayed. The pathological changes, protein expression of endothelial NO synthase (eNOS), phosphorylated eNOS (p-eNOS) and ER stress-related genes in the thoracic aorta were measured. Human umbilical vein endothelial cells (HUVEC) were treated with serum-medicated DJC and then with to detect the reactive species (ROS) levels and C/EBP homologous protein (CHOP) distribution, expression of p-eNOS and ER stress-related genes. DJC therapy exhibited a potent antioxidant activity and effectively prevented the endothelial dysfunction (ED) manifested by promoting p-eNOS expression and enhancing NO release, decreasing lipid deposition (Oil-red O staining, CPT1b and ACC) and inflammation (IL-1β, TNFα, CD68 and p-JNK), alleviating oxidative and ER stress, and decreasing the apoptosis of endothelial cells (TUNEL, BCL-2 and BAX) induced by HFD and respectively. These findings suggest that the administration of DJC had endothelial protective effects against HFD-induced ED, inflammation and apoptosis by alleviating lipid deposition, oxidative and ER stress.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: oxygen

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: oxygen

induces ceramide accumulation, mitochondrial protein hyperacetylation, and mitochondrial dysfunction in porcine oocytes.

Low oocyte quality is a possible causal factor of obesity-induced infertility. High (PA) concentration in follicular fluid is a crucial feature noted in obese women. This study examined how high PA concentration reduced mitochondrial quality in oocytes and investigated a possible countermeasure against mitochondrial dysfunction. Cumulus cell-oocyte complexes were obtained from the ovaries of gilts, and incubated in medium containing PA (0.5 mM) or vehicle (BSA) for 44 h. Culturing oocytes at high PA concentration induced mitochondrial dysfunction determined by high reactive species and low ATP content in oocytes. Furthermore, high PA levels increased mitochondrial acetylation levels determined by a high degree of co-localization of TOMM20 and acetylated-lysine. In addition, high PA levels reduced the expression of Sirtuin 3 (SIRT3) and phosphorylated AMP-activated protein kinase (AMPK), while the AMPK activator, AICAR, restored mitochondrial function as well as oocyte ability and reduced the acetylation of mitochondrial protein. Supplementation of culture medium with dorsomorphin dihydrochloride (an AMPK inhibitor) reduced mitochondrial function and increased mitochondrial protein acetylation. Treatment of oocytes with LB100 (an inhibitor of AMPK dephosphorylation) reduced mitochondrial acetylation levels and restored mitochondrial function. Furthermore, high PA levels increased ceramide accumulation in oocytes, and addition of ceramide to the culture medium also induced mitochondrial dysfunction and increased mitochondrial acetylation. This detrimental effect of ceramide was diminished by AICAR treatment of oocytes. Our results indicated that PA induces ceramide accumulation and downregulates the AMPK/SIRT3 pathway causing mitochondrial protein hyperacetylation and dysfunction in oocytes.

Keyword: oxygen

Metabolic responses of Ulva compressa to single and combined heavy metals.

Accumulation of metals and metabolic responses were studied for two Cd and Cu concentrations (1 and 10\u202fμM) either alone or as a combination in marine macroalga after 7 days of exposure. Cd accumulated more at a low dose (115\u202fμg of Cd/g DW) but Cu at a high dose (378\u202fμg of Cu/g DW); Cu suppressed Cd accumulation (by 57%). Na and Zn levels were unaffected, but higher metal doses depleted K and Ca levels. Higher metal concentrations strongly stimulated reactive species and depleted nitric oxide (NO) formation, but differences between the action of Cd and Cu were not extensive. Higher metal doses increased cell wall thickness with a potential relation to NO signal that is visible mainly in the apoplast in those treatments. A higher Cu dose depleted proline, ascorbic , and phenol levels more than Cd, whereas Cd elevated nonprotein thiols and ascorbic in combined treatments. An eventual role of malic or citric in metal chelation was not evident: malic level decreased in all treatments. The total content of fatty acids reached 16.7\u202fmg/g DW in control with the quantitative order of PUFAs\u202f>\u202fSFAs\u202f>\u202fMUFAs; , vaccenic, linoleic, and α-linolenic acids were the major compounds. Cu was more toxic for fatty acids than Cd (even at 1\u202fμM); mainly, PUFA levels strongly decreased (from 43% of total acids in control to 28.9% and 5.4% at 1 and 10\u202fμM Cu treatment, respectively). Results are precisely and critically discussed in relation to limited literature focused on macroalgae, and a comparison with microalgae is also provided.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: oxygen

Inhibition of inflammation and oxidative stress by an imidazopyridine derivative X22 prevents heart injury from obesity.

Inflammation and oxidative stress plays an important role in the development of obesity-related complications and cardiovascular disease. Benzimidazole and imidazopyridine compounds are a class of compounds with a variety of activities, including anti-inflammatory, antioxidant and anti-cancer. X22 is an imidazopyridine derivative we synthesized and evaluated previously for anti-inflammatory activity in lipopolysaccharide-stimulated macrophages. However, its ability to alleviate obesity-induced heart injury via its anti-inflammatory actions was unclear. This study was designed to evaluate the cardioprotective effects of X22 using cell culture studies and a high-fat diet rat model. We observed that treatment in cardiac-derived H9c2 cells induced a significant increase in reactive species, inflammation, apoptosis, fibrosis and hypertrophy. All of these changes were inhibited by treatment with X22. Furthermore, oral administration of X22 suppressed high-fat diet-induced oxidative stress, inflammation, apoptosis, hypertrophy and fibrosis in rat heart tissues and decreased serum lipid concentration. We also found that the anti-inflammatory and anti-oxidative actions of X22 were associated with Nrf2 activation and nuclear factor-kappaB (NF-κB) inhibition, respectively, both in vitro and in vivo. The results of this study indicate that X22 may be a promising cardioprotective agent and that Nrf2 and NF-κB may be important therapeutic targets for obesity-related complications.© 2016 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: oxygen

Necroptosis is a key pathogenic event in human and experimental murine models of non-alcoholic steatohepatitis.

Hepatocyte cell death, inflammation and oxidative stress constitute key pathogenic mechanisms underlying non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the role of necroptosis in human and experimental NAFLD and its association with tumour necrosis factor α (TNF-α) and oxidative stress. Serum markers of necrosis, liver receptor-interacting protein 3 (RIP3) and phosphorylated mixed lineage kinase domain-like (MLKL) were evaluated in control individuals and patients with NAFLD. C57BL/6 wild-type (WT) or RIP3-deficient (RIP3(-/-)) mice were fed a high-fat choline-deficient (HFCD) or methionine and choline-deficient (MCD) diet, with subsequent histological and biochemical analysis of hepatic damage. In primary murine hepatocytes, necroptosis and oxidative stress were also assessed after necrostatin-1 (Nec-1) treatment or RIP3 silencing. We show that circulating markers of necrosis and TNF-α, as well as liver RIP3 and MLKL phosphorylation were increased in NAFLD. Likewise, RIP3 and MLKL protein levels and TNF-α expression were increased in the liver of HFCD and MCD diet-fed mice. Moreover, RIP3 and MLKL sequestration in the insoluble protein fraction of NASH (non-alcoholic steatohepatitis) mice liver lysates represented an early event during stetatohepatitis progression. Functional studies in primary murine hepatocytes established the association between TNF-α-induced RIP3 expression, activation of necroptosis and oxidative stress. Strikingly, RIP3 deficiency attenuated MCD diet-induced liver injury, steatosis, inflammation, fibrosis and oxidative stress. In conclusion, necroptosis is increased in the liver of NAFLD patients and in experimental models of NASH. Further, TNF-α triggers RIP3-dependent oxidative stress during hepatocyte necroptosis. As such, targeting necroptosis appears to arrest or at least impair NAFLD progression.© 2015 Authors; published by Portland Press Limited.

Keyword: oxygen

Baicalin protects AML-12 cells from lipotoxicity via the suppression of ER stress and TXNIP/NLRP3 inflammasome activation.

Baicalin (BA) is an active flavonoid compound originating from the root of Scutellaria baicalensis Georgi that has been reported to exert anti-inflammation and anti-oxidant effects in liver diseases with a long history. However, its protective and regulatory mechanisms on (PA) induced hepatocyte lipotoxicity remain elusive. In the present work, we investigated the cytoprotective effects of BA in AML-12 hepatocytes against lipotoxicity by inhibiting ER stress, oxidative stress and apoptosis. Our results demonstrated that ER stress was induced by 400\xa0μM\xa0PA in AML-12\xa0cells with elevated expression of ER stress marker IRE1α hyperphosphorylation. BA at 12.5\xa0μM and 25\xa0μM effectively inhibited the expression of p-IRE1α as TUDCA. Flow cytometry analysis and immunofluorescence staining revealed that PA-induced ROS and cell apoptosis were reversed by BA. Furthermore, western blotting revealed that PA-challenged expressions of TXNIP and NLRP3 were dramatically suppressed by BA and TUDCA, suggesting that BA inhibited ER stress through a TXNIP/NLRP3 pathway. Overall, our results indicate that BA alleviates PA-induced cytotoxicity in AML-12\xa0cells via suppression of ER stress and TXNIP/NLRP3 inflammasome activation. These results provide a possible basis of the underlying mechanism and a new insight into the application for BA in the treatment of NAFLD.Copyright © 2017. Published by Elsevier B.V.

Keyword: oxygen

Crucial Roles of 5-HT and 5-HT2 Receptor in Diabetes-Related Lipid Accumulation and Pro-Inflammatory Cytokine Generation in Hepatocytes.

Previously, we confirmed that liver-synthesized 5-HT rather than non-liver 5-HT, acting on the 5-HT2 receptor (5-HT2R), modulates lipid-induced excessive lipid synthesis (ELS). Here, we further revealed the effects of the hepatocellular 5-HT system in diabetes-related disorders.Studies were conducted in male ICR mice, human HepG2 cells, and primary mouse hepatocytes (PMHs) under gene or chemical inhibition of the 5-HT system, key lipid metabolism, and inflammation-related factors. Protein and messenger RNA expression and levels of the factors were determined via western blotting, reverse transcription PCR, and quantitative assay kits, respectively. Hepatic steatosis with inflammation and fibrosis, intracellular lipid droplet accumulation (LDA), and reactive species (ROS) location were determined via hematoxylin and eosin, Masson\'s trichrome, Oil red O, and fluorescent-specific staining, respectively. induced the activation of the 5-HT system: the activation of 5-HT2R, primarily 5-HT2AR, in addition to upregulating monoamine oxidase A (MAO-A) expression and 5-HT synthesis, by activating the G protein/ phospholipase C pathway modulated PKCε activation, resulting in ELS with LDA; the activation of NF-κB, which mediates the generation of pro-inflammatory cytokines, was primarily due to ROS generation in the mitochondria induced by MAO-A-catalyzed 5-HT degradation, and secondarily due to the activation of PKCε. These effects of the 5-HT system were also detected in - or high glucose-treated PMHs and regulated multiple inflammatory signaling pathways. In diabetic mice, co-treatment with antagonists of both 5-HT synthesis and 5-HT2R significantly abolished hepatic steatosis, inflammation, and fibrosis as well as hyperglycemia and dyslipidemia.Activation of the hepatocellular 5-HT system plays a crucial role in inducing diabetes-related hepatic dysfunction and is a potential therapeutic target.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: oxygen

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect inflammation and lipid content in the liver.The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and inflammation induced by HF diet , leading to improved metabolic disorder and associated liver injury in a mouse model of non-alcoholic fatty liver disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: oxygen

Myristic potentiates -induced lipotoxicity and steatohepatitis associated with lipodystrophy by sustaning de novo ceramide synthesis.

(PA) induces hepatocyte apoptosis and fuels de novo ceramide synthesis in the endoplasmic reticulum (ER). Myristic (MA), a free fatty highly abundant in copra/palmist oils, is a predictor of nonalcoholic steatohepatitis (NASH) and stimulates ceramide synthesis. Here we investigated the synergism between MA and PA in ceramide synthesis, ER stress, lipotoxicity and NASH. Unlike PA, MA is not lipotoxic but potentiated PA-mediated lipoapoptosis, ER stress, caspase-3 activation and cytochrome c release in primary mouse hepatocytes (PMH). Moreover, MA kinetically sustained PA-induced total ceramide content by stimulating dehydroceramide desaturase and switched the ceramide profile from decreased to increased ceramide 14:0/ceramide16:0, without changing medium and long-chain ceramide species. PMH were more sensitive to equimolar ceramide14:0/ceramide16:0 exposure, which mimics the outcome of PA plus MA treatment on ceramide homeostasis, than to either ceramide alone. Treatment with myriocin to inhibit ceramide synthesis and tauroursodeoxycholic to prevent ER stress ameliorated PA plus MA induced apoptosis, similar to the protection afforded by the antioxidant BHA, the pan-caspase inhibitor z-VAD-Fmk and JNK inhibition. Moreover, ruthenium red protected PMH against PA and MA-induced cell death. Recapitulating in vitro findings, mice fed a diet enriched in PA plus MA exhibited lipodystrophy, hepatosplenomegaly, increased liver ceramide content and cholesterol levels, ER stress, liver damage, inflammation and fibrosis compared to mice fed diets enriched in PA or MA alone. The deleterious effects of PA plus MA-enriched diet were largely prevented by in vivo myriocin treatment. These findings indicate a causal link between ceramide synthesis and ER stress in lipotoxicity, and imply that the consumption of diets enriched in MA and PA can cause NASH associated with lipodystrophy.

Keyword: oxygen

Oxidative stress induced by modulates K2.3 channels in vascular endothelium.

Elevated plasma free fatty acids level has been implicated in the development of insulin resistance, inflammation, and endothelial dysfunction in diabetic and nondiabetic individuals. However, the underlying mechanisms still remain to be defined. Herein, we investigated the effect of (PA), the most abundant saturated fatty in the human body, on small-conductance Ca-activated potassium channels (K2.3)-mediated relaxation in rodent resistance arteries and the underlying molecular mechanism. The effect of PA on K2.3 in endothelium was evaluated using real-time PCR, Western blotting, whole-cell patch voltage-clamp, wire and pressure myograph system, and reactive species (ROS) were measured by using dihydroethidium and 2\', 7\'-dichlorofluorescein diacetate. K2.3-mediated vasodilatation responses to acetylcholine and NS309 (agonist of K2.3 and K3.1) were impaired by incubation of normal mesenteric arteries with 100\u202fμM\u202fPA for 24\u202fh. In cultured human umbilical vein endothelial cells (HUVECs), PA decreased K2.3 current and expression at mRNA and protein levels. Incubation with the NADPH oxidase (Nox) inhibitor dibenziodolium (DPI) partly inhibited the PA-induced ROS production and restored K2.3 expression. Inhibition of either p38-MAPK or NF-κB using specific inhibitors (SB203580, SB202190 or Bay11-7082, pyrrolidinedithiocarbamate) attenuated PA-induced downregulation of K2.3 and inhibition of p38-MAPK also attenuated PA-induced phosphorylation of NF-κB p65. Furthermore, DPI reversed the increment of phospho-p38-MAPK by PA. These results demonstrated that PA downregulated K2.3 expressions via Nox/ROS/p38-MAPK/NF-κB signaling leading to endothelial vasodilatory dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: oxygen

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro diabetes research.

The interplay between hyper-glycemia and -lipidemia in diabetes mellitus (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for lipid transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: oxygen

High-fat diet promotes experimental colitis by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental colitis in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental colitis model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe colitis compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) tight junction regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes colitis by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes colitis in a 2,4,6-trinitrobenzenesulfonic -induced experimental colitis model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: oxygen

New insights into salvianolic A action: Regulation of the TXNIP/NLRP3 and TXNIP/ChREBP pathways ameliorates HFD-induced NAFLD in rats.

Salvianolic A (SalA), one of the most efficacious polyphenol compounds extracted from Radix Salvia miltiorrhiza (Danshen), has been shown to possess many potential pharmacological activities. This study aimed to investigate whether SalA has hepatoprotective effects against high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) and to further explore the mechanism underlying this process. SalA treatment significantly attenuated HFD-induced obesity and liver injury, and markedly decreased lipid accumulation in HFD-fed rat livers. Moreover, SalA treatment ameliorated HFD-induced hepatic inflammation and oxidative stress by decreasing hepatotoxic levels of cytokines, suppressing the overproduction of reactive species (ROS) and methane dicarboxylic aldehyde (MDA) and preventing the decreased expression of superoxide dismutase (SOD). Importantly, SalA reversed the HFD- or (PA)-induced activation of the NLRP3 inflammasome, the nuclear translocation of ChREBP and the up-regulation of FAS, and these effects were accompanied by TXNIP down-regulation. However, TXNIP siRNA treatment partially abrogated the above-mentioned effects of SalA in PA-treated HepG2 cells. Together, our results demonstrated, for the first time, that SalA protects against HFD-induced NAFLD by ameliorating hepatic lipid accumulation and inflammation, and these protective effects may partially due to regulation of the TXNIP/NLRP3 and TXNIP/ChREBP pathways.

Keyword: oxygen

Hydroxyhydroquinone impairs fat utilization in mice by reducing nitric oxide availability.

Habitual consumption of chlorogenic compounds (CGAs) from coffee increases fat catabolism and reduces body fat; however, the contribution of roasted coffee remains unclear. Hydroxyhydroquinone (HHQ) impairs the vasodilatory and antihypertensive effects of CGAs by reducing nitric oxide (NO) bioavailability. Since HHQ also reduces fat catabolism, we hypothesized that HHQ does so by decreasing NO availability. Therefore, we investigated the effect of HHQ on energy metabolism in KKAy mice. In HHQ-treated mice, fat oxidation was significantly low and dose-dependent, serum and urinary hydrogen peroxide were high, and plasma NO metabolites and S-nitrosylated liver proteins were low. In HHQ-treated mouse hepatocytes, the palmitate-induced increase in cellular consumption was negatively affected, and HHQ or L-NAME reduced cellular fatty utilization. In conclusion, HHQ can impair fat utilization by reducing NO availability in mice. Protein S-nitrosylation reduction in liver cells after HHQ consumption may be associated with impaired fatty oxidation.

Keyword: oxygen

Cultivation of freshwater microalgae in biodiesel wash water.

Biodiesel wash water is a contaminating industrial effluent that must be treated prior to disposal. The use of this effluent as a low-cost alternative cultivation medium for microalgae could represent a viable supplementary treatment. We cultivated 11 microalgae species with potential use for biodiesel production to assess their growth capacities in biodiesel industrial washing waters. Only Monoraphidium contortum, Ankistrodesmus sp., Chlorococcum sp., and one unidentified Chlorophyceae species grew effectively in that effluent. M. contortum showed the highest growth capacity and had the second highest fatty content (267.9\xa0mg\xa0g of DW), predominantly producing (20.9%), 7,10,13-hexadecatrienoic (14%), oleic (16.2%), linoleic (10.5%), and linolenic acids (23.2%). In the second phase of the experiment, the microalgae were cultivated in biodiesel wash water at 75% of its initial concentration as well as in WC (control) medium. After 21\xa0days of cultivation, 25.8 and 7.2% of the effluent nitrate and phosphate were removed, respectively, and the chemical demand was diminished by 31.2%. These results suggest the possibility of cultivating biodiesel producing microalgae in industrial wash water effluents.

Keyword: oxygen

Bromide alleviates fatty -induced lipid accumulation in mouse primary hepatocytes through the activation of PPARα signals.

Increased plasma free fatty acids (FFAs) and liver triglyceride (TG) accumulations have been implicated in the pathogenesis of hepatic steatosis. On the other hand, trace elements function as essential cofactors that are involved in various biochemical processes in mammals, including metabolic homeostasis. Notably, clinical and animal studies suggest that the plasma levels of bromide negatively correlate with those of TG, total cholesterol (TC) and high-density lipoprotein-cholesterol (HDL-C). However, the effect of bromide on lipid accumulation and the direct molecular target responsible for its action remains unknown. Oil red O (ORO) and Nile red staining were used to detect the effect of bromide on lipid accumulation in mouse primary hepatocytes (PHs) treated with different doses of sodium bromide (NaBr) in the presence of FFAs (0.4\xa0mM oleate/ 1:1). Spectrophotometric and fluorometric analyses were performed to assess cellular TG concentrations and rates of fatty oxidation (FAO), respectively, in mouse PHs. We found that bromide decreased FFA-induced lipid accumulation and increased FFA-inhibited consumptions in mouse PHs in a dose-dependent manner via activation of PPARα. Mechanical studies demonstrated that bromide decreased the phosphorylation levels of JNK. More importantly, the PPARα-specific inhibitor GW6471 partially abolished the beneficial effects of bromide on mouse PHs. Bromide alleviates FFA-induced excessive lipid storage and increases rates of FAO through the activation of PPARα/JNK signals in mouse PHs. Therefore, bromide may serve as a novel drug in the treatment of hepatic steatosis.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: oxygen

Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4-MD2 complex.

Reactive species (ROS) contribute to the development of non-alcoholic fatty liver disease. ROS generation by infiltrating macrophages involves multiple mechanisms, including Toll-like receptor 4 (TLR4)-mediated NADPH oxidase (NOX) activation. Here, we show that palmitate-stimulated CD11bF4/80 hepatic infiltrating macrophages, but not CD11bF4/80 Kupffer cells, generate ROS via dynamin-mediated endocytosis of TLR4 and NOX2, independently from MyD88 and TRIF. We demonstrate that differently from LPS-mediated dimerization of the TLR4-MD2 complex, palmitate binds a monomeric TLR4-MD2 complex that triggers endocytosis, ROS generation and increases pro-interleukin-1β expression in macrophages. Palmitate-induced ROS generation in human CD68CD14 macrophages is strongly suppressed by inhibition of dynamin. Furthermore, Nox2-deficient mice are protected against high-fat diet-induced hepatic steatosis and insulin resistance. Therefore, endocytosis of TLR4 and NOX2 into macrophages might be a novel therapeutic target for non-alcoholic fatty liver disease.

Keyword: oxygen

Epigallocatechin gallate improves insulin resistance in HepG2 cells through alleviating inflammation and lipotoxicity.

High levels of circulating free fatty acids (FFAs), inflammation and oxidative stress are important causes for insulin resistance (IR) and type 2 diabetes mellitus. The aim of this study was to investigate the mechanisms of EGCG in alleviating IR in HepG2 cells.HepG2 cells were treated with 25\u202fmM glucose, 0.25\u202fmM (PA), or 50\u202fμM EGCG for 24\u202fh.EGCG increased glucose uptake and decreased glucose content. EGCG markedly decreased the levels of inflammatory and oxidative stress factors including nuclear factor κB (NF-κB), tumor necrosis factor-α, interleukin-6, reactive species, malondialdehyde and p53 protein, and markedly increased superoxide dismutases (SOD), glutathione peroxidase and SOD2 protein. EGCG significantly downregulated the levels of FFAs, triacylglycerol and cholesterol in HepG2 cells. The glucose transporter 2 (GLUT2) protein and its downstream proteins peroxisome proliferator-activated receptor γ coactivator (PGC)-1β were significantly increased, and sterol regulatory element-binding-1c (SREBP-1c) protein, and fatty synthase (FAS) were significantly decreased by EGCG in HepG2. Moreover, the foregoing effects were reversed by siRNA-mediated knockdown of GLUT2.Our data demonstrated that EGCG improved IR, possibly through ameliorating glucose (25\u202fmM) and PA (0.25\u202fmM)-induced inflammation, oxidative stress, and FFAs via the GLUT2/PGC-1β/SREBP-1c/FAS pathway in HepG2 cells.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: oxygen

Berberine Protects Glomerular Podocytes via Inhibiting Drp1-Mediated Mitochondrial Fission and Dysfunction.

Elevated levels of plasma free fatty (FFA) and disturbed mitochondrial dynamics play crucial roles in the pathogenesis of diabetic kidney disease (DKD). However, the mechanisms by which FFA leads to mitochondrial damage in glomerular podocytes of DKD and the effects of Berberine (BBR) on podocytes are not fully understood. : Using the db/db diabetic mice model and cultured mouse podocytes, we investigated the molecular mechanism of FFA-induced disturbance of mitochondrial dynamics in podocytes and testified the effects of BBR on regulating mitochondrial dysfunction, podocyte apoptosis and glomerulopathy in the progression of DKD. : Intragastric administration of BBR for 8 weeks in db/db mice significantly reversed glucose and lipid metabolism disorders, podocyte damage, basement membrane thickening, mesangial expansion and glomerulosclerosis. BBR strongly inhibited podocyte apoptosis, increased reactive species (ROS) generation, mitochondrial fragmentation and dysfunction both and . Mechanistically, BBR could stabilize mitochondrial morphology in podocytes via abolishing (PA)-induced activation of dynamin-related protein 1 (Drp1). : Our study demonstrated for the first time that BBR may have a previously unrecognized role in protecting glomerulus and podocytes via positively regulating Drp1-mediated mitochondrial dynamics. It might serve as a novel therapeutic drug for the treatment of DKD.

Keyword: oxygen

A hypothetical model to solve the controversy over the involvement of UCP2 in palmitate-induced β-cell dysfunction.

The aim of this article is to solve an existing controversy over the involvement of uncoupling protein-2 in the impairment of glucose-stimulated insulin secretion induced by chronic exposure of β-cells to palmitate. We analyzed and compared the results of studies that support and that deny the involvement of uncoupling protein-2 in this impairment. We observed that this impairment could occur in multiple stages. We provide a model in which palmitate-induced impairment of glucose-stimulated insulin secretion is proposed to occur in two stages, early stage and late stage, depending on the integrity of electron supply (glycolysis and Krebs cycle) and transport system through electron transport chain after palmitate treatment. Prolonged exposure of β-cells to palmitate can impair this system. Early-stage impairment occurs due to uncoupling by uncoupling protein-2 when this system is still intact. When this system becomes impaired, late-stage impairment occurs mainly due to reduced glucose-stimulated adenosine triphosphate production independent of uncoupling by uncoupling protein-2. The change in glucose-stimulated uptake after palmitate treatment reflects the integrity of this system and can be used to differentiate between the two stages. Some β-cells lines and islets appear to be more resistant to palmitate-induced impairment of electron supply and transport system than others, and therefore early stage is prominent in the more resistant cell lines and less prominent or absent in the less resistant cell lines. This may help to resolve the pathogenesis of diabetes and to monitor the progression of palmitate-induced β-cell dysfunction.

Keyword: oxygen

Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis.

Autophagy, which is critical for the proper turnover of organelles such as endoplasmic reticulum and mitochondria, affects diverse aspects of metabolism, and its dysregulation has been incriminated in various metabolic disorders. However, the role of autophagy of myeloid cells in adipose tissue inflammation and type 2 diabetes has not been addressed. We produced mice with myeloid cell-specific deletion of Atg7 (autophagy-related 7), an essential autophagy gene (Atg7 conditional knockout [cKO] mice). While Atg7 cKO mice were metabolically indistinguishable from control mice, they developed diabetes when bred to ob/w mice (Atg7 cKO-ob/ob mice), accompanied by increases in the crown-like structure, inflammatory cytokine expression and inflammasome activation in adipose tissue. Mφs (macrophages) from Atg7 cKO mice showed significantly higher interleukin 1 β release and inflammasome activation in response to a plus lipopolysaccharide combination. Moreover, a decrease in the NAD(+):NADH ratio and increase in intracellular ROS content after treatment with in combination with lipopolysaccharide were more pronounced in Mφs from Atg7 cKO mice, suggesting that mitochondrial dysfunction in autophagy-deficient Mφs leads to an increase in lipid-induced inflammasome and metabolic deterioration in Atg7 cKO-ob/ob mice. Atg7 cKO mice were more susceptible to experimental colitis, accompanied by increased colonic cytokine expression, T helper 1 skewing and systemic bacterial invasion. These results suggest that autophagy of Mφs is important for the control of inflammasome activation in response to metabolic or extrinsic stress, and autophagy deficiency in Mφs may contribute to the progression of metabolic syndrome associated with lipid injury and colitis.

Keyword: oxygen

Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon source.

Industrial biotechnology that is able to provide environmentally friendly bio-based products has attracted more attention in replacing petroleum-based industries. Currently, most of the carbon sources used for fermentation-based bioprocesses are obtained from agricultural commodities that are used as foodstuff for human beings. Lignocellulose-derived sugars as the non-food, green, and sustainable alternative carbon sources have great potential to avoid this dilemma for producing the renewable, bio-based hydrocarbon fuel precursors, such as microbial lipid. Efficient bioconversion of lignocellulose-based sugars into lipids is one of the critical parameters for industrial application. Therefore, the\xa0fed-batch cultivation, which is a common method used in industrial applications, was investigated to achieve a high cell\xa0density culture along with high lipid yield and productivity.In this study, several fed-batch strategies were explored to improve lipid production using lignocellulosic hydrolysates derived from corn stover. Compared to the batch culture giving a lipid yield of 0.19\xa0g/g, the dissolved--stat feeding mode increased the lipid yield to 0.23\xa0g/g and the lipid productivity to 0.33\xa0g/L/h. The pulse feeding mode further improved lipid productivity to 0.35\xa0g/L/h and the yield to 0.24\xa0g/g. However, the highest lipid yield (0.29\xa0g/g) and productivity (0.4\xa0g/L/h) were achieved using an automated online sugar control feeding mode, which gave a dry cell weight of 54\xa0g/L and lipid content of 59\xa0% (w/w). The major fatty acids of the lipid derived from lignocellulosic hydrolysates were predominately and oleic , which are similar to those of conventional oilseed plants.Our results suggest that the fed-batch feeding strategy can strongly influence the lipid production. The online sugar control feeding mode was the most appealing strategy for high cell density, lipid yield, and lipid productivity using lignocellulosic hydrolysates as the sole carbon source.

Keyword: oxygen

Fatty levels alterations in THP-1 macrophages cultured with lead (Pb).

As cardiovascular events are one of the main causes of death in developed countries, each factor potentially increasing the risk of cardiovascular disease deserves special attention. One such factor is the potentially atherogenic effect of lead (Pb) on lipid metabolism, and is significant in view of the still considerable Pb environmental pollution and the non-degradability of Pb compounds.Analysis of saturated fatty acids (SFA) (caprylic (C8:0), decanoic (C10:0), lauric (C12:0), tridecanoic (C13:0), myristic (C14:0), pentadecanoic (C15:0), (C16:0), heptadecanoic (C17:0), stearic (C18:0), and behenic (C22:0)), monounsaturated fatty (MUFA) (palmitoleic (C16:1), oleic (18:1w9), trans-vaccenic (C18:1 trans11)), and polyunsaturated fatty (PUFA) (linoleic (C18:2n6), gamma-linolenic (C18:3n6), arachidonic (C20:4n6)), was conducted by gas chromatography. Analysis of stearoyl-CoA desaturase (SCD), fatty desaturase 1 (FADS1) and fatty desaturase 2 (FADS2) expression was performed using qRT-PCR. Oxidative stress intensity (malondialdehyde - MDA concentration) was measured using spectrophotometric method. Intracellular generation of reactive species (ROS) in macrophages was visualized by fluorescence microscopy and quantitatively measured by plate reader.Pb caused quantitative alterations in FAs profile in macrophages; the effect was Pb-concentration dependent and selective (i.e. concerned only selected FAs). In general, the effect of Pb was biphasic, with Pb levels of 1.25\u2009μg/dL and 2.5\u2009μg/dL being stimulatory, and 10\u2009μg/dL being inhibitory on concentrations of selected FAs. The most potent Pb concentration, resulting in increase in levels of 9 FAs, was 2.5\u2009μg/dL, the Pb-level corresponding to the mean blood Pb concentrations of people living in urban areas not contaminated by Pb. Pb was found to exert similar, biphasic effect on the expression of FADS1. However, Pb decreased, in a concentration-dependent manner, the expression of SCD and FADS2. Pb significantly increased MDA and ROS concentration in macrophages.Environmental Pb exposure might be a risk factor resulting in alterations in FAs levels, oxidative stress and increased MDA concentration in macrophages, which might lead to the formation of foam cells and to inflammatory reactions.Copyright © 2019 Elsevier GmbH. All rights reserved.

Keyword: oxygen

Epoxy-Based Organogels for Thermally Reversible Light Scattering Films and Form-Stable Phase Change Materials.

Alkyl chains of β-hydroxyesters synthesized by the capping of terminal epoxy groups of diglycidylether of bisphenol A (DGEBA) with (C16), stearic (C18), or behenic (C22) fatty acids self-assemble forming a crystalline phase. Above a particular concentration solutions of these esters in a variety of solvents led to supramolecular (physical) gels below the crystallization temperature of alkyl chains. A form-stable phase change material (FS-PCM) was obtained by blending the ester derived from behenic with eicosane. A blend containing 20 wt % ester was stable as a gel up to 53 °C and exhibited a heat storage capacity of 161 J/g, absorbed during the melting of eicosane at 37 °C. Thermally reversible light scattering (TRLS) films were obtained by visible-light photopolymerization of poly(ethylene glycol) dimethacrylate-ester blends (50 wt %) in the gel state at room temperature. The reaction was very fast and not inhibited by . TRLS films consisted of a cross-linked methacrylic network interpenetrated by the supramolecular network formed by the esters. Above the melting temperature of crystallites formed by alkyl chains, the film was transparent due to the matching between refractive indices of the methacrylic network and the amorphous ester. Below the crystallization temperature, the film was opaque because of light dispersion produced by the organic crystallites uniformly dispersed in the material. Of high significance for application was the fact that the contrast ratio did not depend on heating and cooling rates.

Keyword: oxygen

Omega-3 fatty acids control productions of superoxide and nitrogen oxide and insulin content in INS-1E cells.

Omega-3 fatty acids have multiple effects in peripheral tissues and pancreatic beta cell function. Dietary depletion of omega-3 fatty acids is associated with pancreatic islet dysfunction and insulin resistance in rats. Herein, the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) on pancreatic beta cell redox state and function were investigated. INS-1E insulin-secreting cells were incubated with EPA and DHA in combination with , and productions of reactive species (ROS), nitric oxide (NO) and insulin were measured. The involvement of the NADPH oxidase complex in ROS production and expression of the antioxidant enzymes was also investigated. After incubation for 1 or 48\xa0h, productions of superoxide (by hydroethidine method), nitric oxide (by 4,5-diaminofluorescein diacetate-DAF-2DA assay), insulin (by radioimmunoassay), and expressions (by western blot analysis) of glutathione peroxidase (GPx-1) and gp91 were measured. EPA and DHA reduced superoxide production after 1-h incubation. After 48\xa0h, reduced superoxide production that was normalized by EPA treatment. increased NO production that was reverted by EPA and DHA. increased insulin secretion after 48\xa0h, whereas both omega-3 fatty acids increased intracellular insulin content. EPA and DHA enhanced GPx-1 expression as well as gp91 glycosylated form. In conclusion, EPA and DHA increased intracellular insulin content and antioxidant enzymatic defense capacity and decreased pro-oxidant generating activities that are associated with maintenance of pancreatic beta cell redox state in response to .

Keyword: oxygen

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving lipid metabolism, antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis (NASH) is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high fat high fructose (HFHF) diet fed C57BL/6J mice, used as model of NASH. The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved lipid homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, mitochondria are intracellular source of reactive species (ROS) and important sites of lipid metabolism, we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti-NASH potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: oxygen

Mutation of IDH1 aggravates the fatty ‑induced oxidative stress in HCT116 cells by affecting the mitochondrial respiratory chain.

Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the metabolic reprogramming of cancer cells; however their functions in lipid metabolism remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic (OA) or (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of mitochondria, immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty ‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.

Keyword: oxygen

, but not high-glucose, induced myocardial apoptosis is alleviated by N‑acetylcysteine due to attenuated mitochondrial-derived ROS accumulation-induced endoplasmic reticulum stress.

Pharmacological inhibition of reactive species (ROS) is a potential strategy to prevent diabetes-induced cardiac dysfunction. This study was designed to investigate precise effects of antioxidant N‑acetylcysteine (NAC) in alleviating diabetic cardiomyopathy (DCM). Echocardiography and histologic studies were performed 12 weeks after streptozocin injection. Protein levels involved in endoplasmic reticulum stress (ERS) and apoptosis were analyzed by western blotting in diabetic hearts or high-glucose (HG, 30\u2009mM)- and (PA, 300\u2009μM)-cultured neonatal rat cardiomyocytes (NRCMs). ROS generation and structural alterations of mitochondria were also assessed. We report that NAC alleviated diabetes-induced cardiac abnormality, including restored ejection fraction (EF %), fraction shortening (FS %), peak E to peak A ratio (E/A) and reduced cardiac hypertrophy and fibrosis. These effects were concomitant with blocked ERS and apoptosis, as evidenced by inactivation of phosphorylated inositol-requiring enzyme-1α (IRE1α)/spliced X-box binding protein 1 (XBP1), phosphorylated protein kinase-like kinase (PERK)/phosphorylated eukaryotic initiation factor 2α (eIF2α) and glucose-regulated protein 78 (GRP78)/activating transcription factor 6 (ATF6α)/C/EBP homologous protein (CHOP) pathways, as well as suppressed Bcl-2-associated X protein (BAX)/B-cell lymphoma-2 (Bcl-2) and cleaved caspase 3 expressions. Mechanistically, PA mediated excessive mitochondrial ROS generation and oxidative stress, which were antagonized by NAC and Mito-TEMPO, a mitochondrial ROS inhibitor. No effects were noted by addition of apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, and NADPH oxidase 4 (NOX 4) and NOX 2 expressions were not altered, indicating that PA-induced ROS generation is independent of NADPH oxidases. Most intriguingly, HG failed to promote ROS production despite its ability to promote ERS and apoptosis in NRCMs. Collectively, these findings indicate that NAC primarily abrogates PA-mediated mitochondrial ROS through ERS and therefore alleviates myocardial apoptosis but has little effect on HG-induced cardiac injury. This uncovers a potential role for NAC in formulating novel cardioprotective strategies in DCM patients.

Keyword: oxygen

Hepatic FTO expression is increased in NASH and its silencing attenuates -induced lipotoxicity.

Non-alcoholic steatohepatitis (NASH) is one of the most common causes of liver failure worldwide. It is characterized by excess fat accumulation, inflammation, and increased lipotoxicity in hepatocytes. Currently, there are limited treatment options for NASH due to lack of understanding of its molecular etiology. In the present study, we demonstrate that the expression of fat mass and obesity associated gene (FTO) is significantly increased in the livers of NASH patients and in a rodent model of NASH. Furthermore, using human hepatic cells, we show that genetic silencing of FTO protects against palmitate-induced oxidative stress, mitochondrial dysfunction, ER stress, and apoptosis in\xa0vitro. Taken together, our results show that FTO may have a deleterious role in hepatic cells during lipotoxic conditions, and strongly suggest that up-regulation of FTO may contribute to the increased liver damage in NASH.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: oxygen

Flavin-Containing Monooxygenase 3 Reduces Endoplasmic Reticulum Stress in Lipid-Treated Hepatocytes.

Isoforms of flavin-containing monooxygenase (FMO) are involved in xenobiotic metabolism but have also been implicated in the regulation of glucose and lipid homeostasis and in the development of atherosclerosis. However, we have recently shown that improved insulin action is associated with increased FMO expression in livers of protein kinase C-deficient mice. Here, we investigated whether FMO3 expression affected insulin signaling, glucose metabolism, and endoplasmic reticulum (ER) stress in hepatocytes. HepG2 and IHH hepatocytes were transfected with FMO3 cDNA for overexpression, or small interfering RNA for knockdown. Cells were treated with palmitate to induce insulin resistance and insulin signaling, phosphoenolpyruvate carboxykinase (PEPCK) gene expression and ER stress markers were examined by immunoblotting and RT-PCR. Glycogen synthesis was measured using [(14)C]glucose. Palmitate treatment reduced insulin signaling at the level of Akt phosphorylation and glycogen synthesis, which were little affected by FMO3 overexpression. However, the fatty also increased the levels of several ER stress markers and activation of caspase 3, which were counteracted by FMO3 overexpression and exacerbated by FMO3 knockdown. Although FMO3 expression did not reverse lipid effects on protein thiol redox in hepatocytes, it did prevent up-regulation of the gluconeogenic enzyme PEPCK by pharmacological ER stress inducers or by palmitate. ER stress and PEPCK levels were also reduced in livers of fat-fed protein kinase Cδ-deficient mice. Our data indicate that FMO3 can contribute to the regulation of glucose metabolism in the liver by reducing lipid-induced ER stress and the expression of PEPCK, independently of insulin signal transduction.

Keyword: oxygen

Effects of a mixture of organisms, Lactobacillus acidophilus or Streptococcus faecalis on delta6-desaturase activity in the livers of rats fed a fat- and cholesterol-enriched diet.

The effect of a mixture of organisms (a probiotic mixture) comprising Bacillus, Lactobacillus, Streptococcus, Clostridium, Saccharomyces, and Candida (10(7-8) colony-forming units/g rice bran of each component) on delta6-desaturase activity in liver microsomes was compared with those of Lactobacillus acidophilus and Streptococcus faecalis. There were four treatment groups. Each group of these rats received rice bran (control), the mixture of organisms, L. acidophilus, or S. faecalis (30 g/kg) along with a fat- and cholesterol-enriched diet for 4 wk. The serum total cholesterol concentration of the group fed the mixture of organisms was reduced by 15-33% compared with the other groups at the end of the 4-wk feeding period (P<0.05). The proportion of in the serum phosphatidylcholine (PC) for the control group was significantly higher than those of the other groups. The proportion of arachidonic in the serum PC for the mixed-organism group was also significantly higher than those of the other groups. The proportion of arachidonic in the liver PC for the mixed-organism group was significantly higher than those of the control and S. faecalis groups. The ratio of arachidonic /linoleic was significantly higher in the liver PC of rats fed the mixed organisms compared with the control group (P<0.05). The delta6-desaturase activity in the liver microsomal fraction of the mixed-organism group was significantly higher than those of the other groups. The delta6-desaturase activity correlated positively with the ratio of arachidonic /linoleic of liver PC, the correlation coefficient (r) being 0.819 (P<0.001). The results indicate that the effect of the mixture of organisms was to increase delta6-desaturase activity and serum arachidonic and decrease cholesterol compared to the other organisms and control, but the mechanism whereby the enzyme activity was related to serum cholesterol does not appear to have been explored.

Keyword: probiotics

Brain and liver fatty composition changes upon consumption of Lactobacillus rhamnosus LA68.

Recent reports suggest that the metabolic activity of the enteric microbiota may influence the fatty composition of the host tissue. There are many studies dealing with the influence of lactobacilli on various pathological conditions, and some of the effects are strain-specific. This study was designed to test the effects of a particular Lactobacillus strain, Lactobacillus rhamnosus LA68 on fatty composition of the liver and the brain of C57BL/6 mice in the absence of an underlying pathological condition. Female mice were supplemented with live L. rhamnosus LA68 bacteria for the duration of 1 month. Serum biochemistry was analyzed and liver and brain fatty composition was assessed by gas-liquid chromatography. Significant changes in liver and brain fatty composition were detected. In the liver tissue we detected an increase in palmitoleic (p\u2009=\u20090.038), while in the brain compartment we found an increase in (p\u2009=\u20090.042), stearic (p\u2009=\u20090.017), arachidonic (p\u2009=\u20090.009) and docosahexaenoic (p\u2009=\u20090.004) for control versus experimental group. These results show discrete changes caused by LA68 strain consumption. Even short duration of administration of LA68 influences the fatty composition of the host which adds to the existing knowledge about Lactobacillus host interaction, and adds to the growing knowledge of metabolic intervention possibilities.

Keyword: probiotics

Effect of dietary Rhodobacter capsulatus on lipid fractions and egg-yolk fatty composition in laying hens.

The present study was conducted to investigate the effect of dietary Rhodobacter capsulatus on lipid fractions and egg-yolk fatty composition in laying hens. Thirty-six laying hens (30\u2003weeks old) were randomly assigned into two dietary groups fed diets with (0.04%) or without (control) R.\xa0capsulatus for a 60-day feeding trial. Dietary R.\xa0capsulatus decreased (p\u2003<\u20030.05) serum and hepatic cholesterol and increased (p\u2003<\u20030.05) the excreta cholesterol, and resultant lower (p\u2003<\u20030.05) cholesterol contents in egg yolk. The concentration of polyunsaturated fatty acids (PUFA) and ratio to saturated fatty acids in egg yolk was improved (p\u2003<\u20030.05) by dietary R.\xa0capsulatus. The concentration of hepatic bile was increased (p\u2003<\u20030.05) and excreta bile was decreased (p\u2003<\u20030.01) in the laying hens fed R.\xa0capsulatus diet. The incorporation of 1-(14) C- into hepatic lipids and lipid fractions was increased (p\u2003<\u20030.05) in laying hens fed R.\xa0capsulatus diet. Moreover, dietary R.\xa0capsulatus did not appear to cause any adverse effects on laying hen performances. Therefore, dietary supplementation of R.\xa0capsulatus in layer diets may be a feasible means of producing eggs with lower cholesterol and higher PUFA contents for health conscious consumers.© 2011 Blackwell Verlag GmbH.

Keyword: probiotics

Potential of Laurencia obtusa as a substrate for the development of a probiotic Saccharomyces cerevisiae.

Laurencia obtusa (Ceramiales, Rhodophyta) has tremendous nutritional value, being high in proteins, oligosaccharides, vitamins, essential minerals, and fatty acids, and it is a rich source of amino acids and trace elements. In this study, L. obtusa was extracted and subjected to phenolic, sugar and flavonoid analyses.The fatty , vitamin and phytosterol contents in Saccharomyces cerevisiae were evaluated when it was incubated with L. obtusa dry biomass. The fatty acids in the lipid extract were analysed after converting them into methyl esters using gas chromatography, and vitamin concentrations were measured using high-performance liquid chromatography (HPLC). According to the achieved results, the total fatty levels and vitamin contents of the S. cerevisiae prepared with algal extract increased at different rates. Our results showed that α-tocopherol decreased in the group in which the S. cerevisiae was added the algal extract. When compared to the control group, ergesterol increased in the group in which L. obtusa extract was added. Additionally, when compared to the control group in which L. obtusa extract was added, stearic (18:0), oleic (18:1) and linoleic (18:2) increased in the other groups. Palmitoleic (16:1) increased in the L. obtusa culture medium, but decreased in the L. obtusa culture medium. In conclusion, it was determined that the L. obtusa extract added to the development medium of S. cerevisiae\xa0caused differences in the synthesis of some vitamins and fatty acids.

Keyword: probiotics

Supplemental flaxseed modulates ovarian functions of weanling gilts via the action of selected fatty acids.

The aim of this study was to examine the influence of dietary flaxseed on the endocrine and ovarian functions of weanling gilts challenged with E. coli and Coronavirus infections treated with dietary probiotic cheeses and to understand the possible mechanisms of its effects on ovarian function. were used as a natural substitution for antibiotics and 10% dietary flaxseed is an effective prebiotic which supports the action of and has other beneficial effects on the organism. with or without flaxseed were fed to weanling gilts starting 10 days before and lasting up until 14 days after weaning. The ovaries were measured and histologically analysed. The blood samples for the levels of steroid hormones and insulin-like growth factor I (IGF-I) were assessed using immunoassays and the levels of fatty acids were assessed using gas chromatography. All samples were collected on the day of weaning and 14 days after weaning. On the day of weaning, increased levels of linoleic and IGF-I was associated with higher body weight. The steroid hormones were not affected by the diet. The conversion of alpha-linolenic (ALA) to timodonic (EPA) and cervonic (DHA) acids were lower compared to controls, and together with high levels of myristic, and palmitoleic acids was associated with the higher proliferation and lower apoptosis in the primordial, primary and secondary follicles; although the inhibition of the cell cycle was observed in relation to the low level of eicosadienoic . The high levels of ALA, EPA and DHA and the low levels of myristic, and palmitoleic acids may have been the effect of flaxseed feeding 14 days post-weaning and may have had a reverse effect on the proliferation and apoptosis of ovarian follicles. These data suggest that flaxseed may suppress the follicle development in weanlings via the stimulation of apoptosis and the inhibition of proliferation via the modulation of the metabolism of selected fatty acids.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: probiotics

Antifungal Attributes of Lactobacillus plantarum MYS6 against Fumonisin Producing Fusarium proliferatum Associated with Poultry Feeds.

Fumonisins, being common in occurrence in maize-based feeds, pose a great threat to animal and human health. The present study is aimed at determining the antifungal activity of Lactobacillus plantarum MYS6 against a fumonisin producing fungus, Fusarium proliferatum MYS9. The isolate was subjected to standard tests for determining its probiotic attributes and antifungal properties. L. plantarum MYS6 thrived well at pH 3.0 and 6.0, and exhibited strong resistance up to 3% bile. The isolate showed a high degree of cell surface hydrophobicity corresponding to its strong adhesion to chicken crop epithelial cells. Co-inoculation with the fungus on modified de Man Rogosa Sharpe medium revealed the inhibitory effect of L. plantarum MYS6 on fungal growth and biomass. Observation using scanning electron microscopy showed distortion of hyphal structures, swollen tips and disrupted conidia. Conidia germination inhibition assay restrained germination and showed deformed hyphae. The bioprotective feature of the isolate was evident by the inhibition of fungal development in maize-kernel treated with the cell free supernatant of L. plantarum MYS6. Both the isolate and its extracellular metabolites lowered fumonisin content in feed model up to 0.505 mg/Kg of feed and 0.3125 mg/Kg of feed respectively when compared to the level of 0.870 mg/Kg of feed in control. The major antifungal compounds produced by the isolate were 10-Octadecenoic , methyl ester; , methyl ester; heptadecanoic , 16-methyl ester; stearic and lauric . L. plantarum MYS6 reduced 61.7% of fumonisin possibly by a binding mechanism. These findings suggest the application of L. plantarum MYS6 as an efficient probiotic additive and biocontrol agent in feed used in poultry industry. Additionally, the antifungal metabolites pose a conspicuous inhibition of Fusarium growth and fumonisin production.

Keyword: probiotics

[Breast milk: biological constituents for health and well-being in infancy].

Many factors are involved in infants' health; one of the most important of them may be the kind of early feeding. Recent evidences suggest that breastfeeding, in addition to its well-established beneficial effects during lactation period, provides also beneficial long-term effects, like the protection against infectious and immune-related diseases, a better cognitive development, a decreased risk of metabolic syndrome and of obesity. It has been reported that the early feeding mode affects growth and body composition and it could be considered a critical factor for metabolic development. Human milk is a source of different nutrients and bioactive factors, especially hormones and growth factors like leptin, ghrelin, insulin, insulin-like growth factor (IGF-I) playing a role in food intake regulation, metabolism and body composition. In particular breast milk leptin may provide a physiological explanation for a number of advantages seen in reaching proper growth and energy balance in breast-fed infants compared with formula fed ones. Etiopathogenesis and therapeutic approach in common minor gastrointestinal diseases in infants are important subject of study for pediatricians. Colic, constipation and regurgitation can be considered feeding problems and they might benefit from dietary treatment. Regarding infantile colic, dietary modifications seem to be more suitable than pharmacological treatment in resolving symptoms; also prebiotics and are useful for this aim. The occurence of constipation is related to the kind of feeding and it is lower in breastfed infants. Moreover formulas with and beta- could promote a regression of symptoms. A dietary approach may be useful also in regurgitation. Anyway we have to remember that breastfeeding require a supplementation of vitamin D and K for some months and a correct weaning program is needed from the 5th-6th months of life to prevent iron deficiency.

Keyword: probiotics

Effect of high β-palmitate content in infant formula on the intestinal microbiota of term infants.

(PA) constitutes 17% to 25% of the human milk fatty acids, and ~70% is esterified in the sn-2 position of triglycerides (β-palmitate). In the sn-2 position, PA is not hydrolyzed and thus is efficiently absorbed. The PA in palm oils, commonly used in infant formulas, is esterified in the sn-1 and sn-3 positions. In these positions, PA is hydrolyzed and forms poorly absorbed calcium complexes. The present study assessed whether high β-palmitate in infant formulas affects the intestinal flora.Thirty-six term infants were enrolled: 14 breast-fed (BF group) and 22 formula-fed infants who were randomly assigned to receive formula containing high β-palmitate (HBP group, n=14), or low β-palmitate (LBP group, n=8), where 44% and 14% of the PA was β-palmitate, respectively. The total amount of PA in the formulas was 19% and 22% in the LBP and HBP groups, respectively. Neither formula contained pre- or . Stool samples were collected at enrollment and at 6 weeks for the quantification of bacteria.At 6 weeks, the HBP and BF groups had higher Lactobacillus and bifidobacteria counts than the LBP group (P<0.01). The Lactobacillus counts at 6 weeks were not significantly different between the HBP and BF groups. Lactobacillus counts were 1.2×10¹⁰, 1.2×10¹¹, and 5.6×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively. Bifidobacteria counts were 5.1×10⁹, 1.2×10¹¹, and 3.9×10¹⁰ CFU/g for LBP, HBP, and BF groups, respectively.HBP formula beneficially affected infant gut microbiota by increasing the Lactobacillus and bifidobacteria counts in fecal stools.ClinicalTrials.gov .

Keyword: probiotics

Comparison of stationary phases in reversed-phase TLC for correlation between structure and biological response of .

A series of C16,C18 and C20 fatty acids and their ethyl esters and alcohols were investigated as possible stationary phases in reversed-phase TLC for the correlation between structure and biological response (antistaphylococcal activity). Ten (omego-amino acids and their L-histidine dipeptides) were used as the biologically active compounds. The mobile phase was 70% acetone in water. The best correlations were obtained with hexadecanoic () or cis9,cis-12,cis-15-octadecatrienoic , 1-hexadecanol or cis-9-octadeconol, and ethyl hexadecanoate for the fatty acids, their alcohols, and their ethyl esters, respectively. Among all compounds, the following relation was obtained: fatty acids = alcohols greater than ethyl esters greater than white paraffin oil.

Keyword: probiotics

Randomized double-blind study of the nutritional efficacy and bifidogenicity of a new infant formula containing partially hydrolyzed protein, a high beta- level, and nondigestible oligosaccharides.

The aim of this study was to evaluate the nutritional efficacy and bifidogenic characteristics of a new infant formula containing partially hydrolyzed whey protein, modified vegetable oil with a high beta- content, prebiotic oligosaccharides, and starch.In a double-blind study, healthy formula-fed term infants aged younger than 2 weeks were randomized to receive either the new infant formula (NF) or a standard formula (SF) until the age of 12 weeks. Anthropometric measurements were taken at enrollment, 6 weeks, and 12 weeks. In a subsample of infants, blood samples were taken at 6 weeks and stool samples were taken at enrollment and 6 weeks. Blood samples were analyzed for biochemical measures of protein status and amino acids, and stools were analyzed for total bacteria and bifidobacteria. Mothers completed a feeding diary and questionnaire at 6 and 10 weeks.One hundred fifty-four infants were enrolled in the study; 102 completed the trial. The growth of infants in both formula groups was in line with published growth curves. During the first 6 weeks, NF girls gained more weight and head circumference than the SF girls. These velocity differences were not maintained throughout the 12-week study period. The NF stools had a higher proportion of bifidobacteria at 6 weeks compared with the SF stools, and they were softer. There were no clinically significant differences in the blood biochemical and amino values between groups. Both formulas were well tolerated by the infants.When compared with a standard infant formula, the new formula supported satisfactory growth, led to higher counts of bifidobacteria in the feces, produced blood bio-chemical values typical of formula-fed infants, and was well tolerated.

Keyword: probiotics

A Metabolomics Approach Reveals Immunomodulatory Effects of Proteinaceous Molecules Derived From Gut Bacteria Over Human Peripheral Blood Mononuclear Cells.

There are strong evidences that influence the immune status of the host, in a strain-specific manner, acting in the gastrointestinal tract. On the hypothesis that certain extracellular proteins and peptides from gut bacteria may mediate part of this immunomodulation and assuming they are able to diffuse through the mucus layer and interact with immune cells we have developed this work. Our study attempts to understand the immunomodulatory mechanisms of (i) Pext, the extracellular protein fraction of DSM20079, (ii) HM14, a peptide encrypted in an extracellular glycoside hydrolase from NCIMB 8809 and (iii) O111:B4 lipopolysaccharide (LPS), a well-known pro-inflammatory molecule, over human peripheral blood mononuclear cells (PBMCs). An untargeted LC-ESI-QTOF-MS metabolomics approach was applied to reveal intracellular changes in treated-PBMCs isolated from healthy donors. Differences in NADH arrest, NAD concentration reduction, as well as increases in and methanephrin were observed in HM14 and Pext treated-cells compared to those stimulated with LPS. This would support an anti-inflammatory molecular mechanism of action of such proteinaceous molecules. Moreover, this methodology has confirms the importance of metabolomics approaches to better understanding immune cell responses to gut bacterial-derived molecules.

Keyword: probiotics

The triglyceride-lowering effect of supplementation with dual probiotic strains, Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032: Reduction of fasting plasma lysophosphatidylcholines in nondiabetic and hypertriglyceridemic subjects.

This study evaluated the triglyceride (TG)-lowering effects of consuming dual probiotic strains of Lactobacillus curvatus (L. curvatus) HY7601 and Lactobacillus plantarum (L. plantarum) KY1032 on the fasting plasma metabolome.A randomized, double-blind, placebo-controlled study was conducted on 92 participants with hypertriglyceridemia but without diabetes. Over a 12-week testing period, the probiotic group consumed 2 g of powder containing 5 × 10(9) colony-forming units (cfu) of L. curvatus HY7601 and 5 × 10(9) cfu of L. plantarum KY1032 each day, whereas the placebo group consumed the same product without . Fasting plasma metabolomes were profiled using UPLC-LTQ-Orbitrap MS. After 12 weeks of treatment, the probiotic group displayed a 20% reduction (p = 0.001) in serum TGs and 25% increases (p=0.001) in apolipoprotein A-V (apoA-V). At the 12-week follow-up assessment, the following 11 plasma metabolites were significantly reduced in the probiotic group than the placebo group: palmitoleamide, amide, oleamide, and lysophosphatidyl choline (lysoPC) containing C14:0, C16:1, C16:0, C17:0, C18:3, C18:2, C18:1, and C20:3. In the probiotic group, changes (▵) in TG were negatively correlated with ▵ apoA-V, which was positively correlated with ▵ FFA. In addition, ▵ FFA was strongly and positively correlated with ▵ lysoPCs in the probiotic group but not the placebo group.The triglyceride-lowering effects of probiotic supplementation, partly through elevated apoA-V, in borderline to moderate hypertriglyceridemic subjects showed reductions in plasma metabolites, fatty primary amides and lysoPCs (; http://www.clinicaltrials.gov). Clinical trials: ; http://www.clinicaltrials.gov.Copyright © 2015 Elsevier B.V. All rights reserved.

Keyword: probiotics

Antibacterial metabolites secreted under glucose-limited environment of the mimicked proximal colon model by lactobacilli abundant in infant feces.

The most abundance of anti-Salmonella lactic bacteria (LAB) was found in feces of naturally born, exclusively breastfed Thai infants. Six strains of Lactobacillus plantarum and one strain of Lactobacillus paracasei were selected and identified. In the co-cultivation assay, L. plantarum subsp. plantarum I62 showed the strongest and broadest antibacterial activity against Escherichia coli, Shigella sonnei, Salmonella Paratyphi A, and Salmonella Typhimurium SA 2093 under the mimicked proximal colon condition, in which glucose and other nutrients were limited. According to GC-MS analysis, the major antibacterial contribution of organic acids secreted by L. plantarum I62 grown in the presence of glucose was dramatically reduced from 95.8 to 41.9\xa0% under glucose-limited niche. The production of low-pK a acids, such as lactic, 1,2-benzenedicarboxylic, and 3-phenyllactic acids, was remarkably dropped. Surprisingly, higher-pK a acids such as 5-chlorobenzimidazole-2-carboxylic, pyroglutamic, , and oleic acids were enhanced. Moreover, cyclic dipeptides, ketones, alkanes, alcohols, and miscellaneous compounds, which were pH-independent antibacterial metabolites, became dominant. The electron microscopy strongly supported the synergistic attacks of the multiple antibacterial components targeting outer and cytoplasmic membranes leading to severe leakage and cell disruption of Salmonella Typhimurium. This strain poses to be a potential probiotic candidate for effectively controlling and treating human foodborne bacterial infection.

Keyword: probiotics

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Prostaglandin precursors in plasma phospholipids of patients with : effects of treatment with coal tar.

Plasma phospholipids of patients with have significantly reduced levels of dihomogammalinolenic (20:3n-6), arachidonic (20:4n-6) and adrenic (22:4n-6), the precursors of the 1, 2 and homo-2 series of prostaglandins (PGs). Concentrations of the 3 series PG precursor, eicosapentaenoic (20:5n-3) were normal. Hospital treatment with a coal tar regime produced a rise in 20:3n-6 to levels which were significantly above normal.

Keyword: psoriasis

Studies on fatty acids in pus of pustulosa palmo-plantaris.

Thin-layer chromatography was used to find out if there were any quantitative differences in fatty content in pus from the palmar pustules in ten patients with pustulosa palmo-plantaris (PPP) before and after long-term tetracycline therapy. No significant differences were found, indicating that it was not possible to correlate the good therapeutic effect of antibiotics reported in PPP with a decrease in the fatty content of pus from the pustles. However, the series was small and investigations are now in progress to establish whether small differences of significance can be demonstrated in a larger series. By combined gas chromatography and mass spectrometry the fatty acids of pus in patients with pustulosa palmo-plantaris were analysed. Stearic and acids were found to be predominant.

Keyword: psoriasis

Free fatty acids in the blood serum of psoriatics.

Free fatty acids in the serum of 60 patients with generalized were studied by a gas-chromatographic method. The same investigations in 40 healthy persons served for control purposes. It was established with statistical probability (p less than 0.001) that in the serum of psoriatics the levels of (C 16: 0), stearic (C 18: 0), arachidic (C 20: 0), and docosadienic (C 20: 2) acids were increased, while the levels of the linoleic (C 18: 2) and arachidonic (C 20: 4) acids were depressed. There is no behenic (C 22: 0) in the blood serum of psoriatics. Thalassotherapy caused no significant changes in the levels of these free fatty acids in psoriatics.

Keyword: psoriasis

A heterocomplex formed by the calcium-binding proteins MRP8 (S100A8) and MRP14 (S100A9) binds unsaturated fatty acids with high affinity.

We show that unsaturated fatty acids (FAs) bind reversibly and with high affinity to a heterocomplex of 34 kDa (FA-p34) formed by the non-covalent association of two calcium-binding proteins of the S100 family: MRP8 (S100A8) and MRP14 (S100A9). Fatty -competition studies on the [3H]oleic .FA-p34-complex show that oleic, alpha-linoleic, gamma-linolenic, and arachidonic acids have IC50 values of about 1 microM, whereas and stearic acids are poor competitors. The binding of arachidonic is saturable with a single class of binding site per FA-p34, and a dissociation constant (Kd) of 0.13 microM is calculated. The individual subunits MRP8 and MRP14 show no binding properties for fatty acids, whereas a p34 complex reconstituted in vitro by the recombinant molecules exhibits binding properties, suggesting that the fatty -binding site of FA-p34 is created through heterocomplex formation. Furthermore, we demonstrate that lowering free Ca2+ levels to 16 nM results in a loss of the fatty -binding capacity of purified FA-p34. In calcium-induced differentiating keratinocytes, the amounts of FA-p34 are increased in the particulate (2.0 +/- 0.5 pmol of [3H]oleic /mg protein) and in the cytosolic (4.5 +/- 0.6 pmol of [3H]oleic /mg protein) fractions, whereas no FA-p34 can be detected in non-differentiated cultured keratinocytes. In abnormally differentiated keratinocytes () and in human polymorphonuclear leukocytes, FA-p34 is highly expressed (31.35 +/- 1.6 and 349.8 +/- 17.9 pmol of [3H]oleic /mg protein, respectively), pointing toward a role for this heteromer in mediating effects of unsaturated fatty acids in a calcium-dependent way during cell differentiation and/or inflammation.

Keyword: psoriasis

Optimization of physiological lipid mixtures for barrier repair.

Three stratum corneum lipids, ceramides, cholesterol (CHOL), and free fatty acids (FA), are required for permeability barrier homeostasis. Recent studies have shown that application of one or two of these lipids to perturbed skin delays barrier recovery; only equimolar mixtures allow normal recovery. We asked here whether any physiological lipid mixtures improve barrier repair, as assessed by transepidermal water loss. Whereas an equimolar ratio of ceramides, CHOL, and FA (either the essential fatty , linoleic , or the nonessential FAs, or stearic acids) allows normal repair, further acceleration of barrier repair occurs as the ratio of any of these ingredients is increased up to 3-fold. Similar preliminary results were obtained in damaged human skin. Likewise, while acylceramides alone delay barrier recovery, acylceramides: CHOL mixtures within a specific range of molar rations dramatically improve barrier repair. Furthermore, glycosyl ceramides, sphingomyelin, and triglycerides substitute effectively for ceramides and FA, respectively, but neither phospholipids nor cholesterol esters substitute for FA and CHOL, respectively. These studies show the specific requirements of selected stratum corneum lipid mixtures for optimized barrier repair in murine skin, with further validation in human skin. Utilization of physiologic lipids according to these parameters could lead to new forms of topical therapy for dermatoses (e.g., , atopic dermatitis, and irritant dermatitis) triggered by abnormal barrier function.

Keyword: psoriasis

Antipsoriatic effect of fatty enriched fraction of Vernonia anthelmintica Willd. fruits.

Vernonia anthelmintica has been utilized conventionally as an ingredient in Ayurveda and traditional Uighur medicine for management of various skin ailments, and scientific data's have substantiated its use in treating vitiligo, dermatosis and leucoderma. The present investigation was focused to evaluate the antipsoriatic activity of V. anthelmintica fruit extracts and fractions.Ointment containing dichloromethane (DCM) and methanol (MeOH) extracts at topical dose of 2.5% and 5% (w/w) was evaluated using mouse tail model of . Bioactivity-guided fractionation (F-F) of most active extract was carried out and fractions were again subjected to mouse tail model. Further the activity of bioactive fraction was confirmed in HaCaT (human keratinocyte) cell line using MTT (3-(4,5-dimethylthiazol-2-yl)-\u202f2,5-diphenyltetrazolium bromide) assay and its chemical characterization was done via gas chromatography mass spectrometry (GC-MS).The dichloromethane extract (5%, w/w) showed statistically significant (*\u202fp\u202f<\u202f0.05) antipsoriatic activity (66.97\u202f±\u202f2.68%) with respect to control (25.45\u202f±\u202f1.80%) and equivalent to that of the standard drug, retino-A 0.05%, (72.47\u202f±\u202f2.14%) in terms of degree of orthokeratosis, whereas methanol extract (5%, w/w) showed significant (*\u202fp\u202f<\u202f0.05) differentiation (45.86\u202f±\u202f2.02%) in comparison to the control group. Out of all fractions, F showed statistically significant (*\u202fp\u202f<\u202f0.05) antipsoriatic activity (69.27\u202f±\u202f2.76%) with respect to control and equivalent to that of the standard. F (15.6-1000\u202fµg/ml) showed dose-dependent inhibition of HaCaT cell lines proliferation which suggests keratinocyte modulating activity of V. anthelmintica. Chemical characterization of F revealed that essential fatty acids (i.e., linoleic , , oleic and stearic ) formed the bulk of bioactive fraction.Ameliorative effect of V. anthelmintica in might be attributed to the presence of essential fatty acids and thus corroborates its traditional use in the treatment of skin ailments.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: psoriasis

Free fatty acids sensitize dendritic cells to amplify TH1/TH17-immune responses.

Obesity is associated with body fat gain and impaired glucose metabolism. Here, we identified both body fat gain in obesity and impaired glucose metabolism as two independent risk factors for increased serum levels of free fatty acids (FFAs). Since obesity is associated with increased and/or delayed resolution of inflammation observed in various chronic inflammatory diseases such as , we investigated the impact of FFAs on human monocyte-derived and mouse bone marrow-derived dendritic cell (DCs) functions relevant for the pathogenesis of chronic inflammation. FFAs such as (PA) and oleic (OA) did not affect the pro-inflammatory immune response of DCs. In contrast, PA and OA sensitize DCs resulting in augmented secretion of TH1/TH17-instructive cytokines upon pro-inflammatory stimulation. Interestingly, obesity in mice worsened a TH1/TH17-driven -like skin inflammation. Strong correlation of the amount of total FFA, PA, and OA in serum with the severity of skin inflammation points to a critical role of FFA in obesity-mediated exacerbation of skin inflammation. Our data suggest that increased levels of FFAs might be a predisposing factor promoting a TH1/TH17-mediated inflammation such as in response to an inflammatory danger signal.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: psoriasis

Free fatty acids: potential proinflammatory mediators in rheumatic diseases.

Due to their role in inflammatory metabolic diseases, we hypothesised that free fatty acids (FFA) are also involved in inflammatory joint diseases. To test this hypothesis, we analysed the effect of FFA on synovial fibroblasts (SF), human chondrocytes and endothelial cells. We also investigated whether the toll-like receptor 4 (TLR4), which can contribute to driving arthritis, is involved in FFA signalling.Rheumatoid arthritis SF, osteoarthritis SF, psoriatic arthritis SF, human chondrocytes and endothelial cells were stimulated in vitro with different FFA. Immunoassays were used to quantify FFA-induced protein secretion. TLR4 signalling was inhibited extracellularly and intracellularly. Fatty translocase (CD36), responsible for transporting long-chain FFA into the cell, was also inhibited.In rheumatoid arthritis synovial fibroblasts (RASF), FFA dose-dependently enhanced the secretion of the proinflammatory cytokine IL-6, the chemokines IL-8 and MCP-1, as well as the matrix-degrading enzymes pro-MMP1 and MMP3. The intensity of the response was mainly dependent on the patient rather than on the type of disease. Both saturated and unsaturated FFA showed similar effects on RASF, while responses to the different FFA varied for human chondrocytes and endothelial cells. Extracellular and intracellular TLR4 inhibition as well as fatty transport inhibition blocked the -induced IL-6 secretion of RASF.The data show that FFA are not only metabolic substrates but may also directly contribute to articular inflammation and degradation in inflammatory joint diseases. Moreover, the data suggest that, in RASF, FFA exert their effects via TLR4 and require extracellular and intracellular access to the TLR4 receptor complex.Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Keyword: psoriasis

DEFICENT CHOLESTEROL ESTERIFYING ABILITY OF LESION-FREE SKIN SURFACES IN PSORIATIC INDIVIDUALS.

Keyword: psoriasis

[Free fatty -determinations on the skin of psoriatics].

Keyword: psoriasis

Co-treatment with retinyl retinoate and a PPARα agonist reduces retinoid dermatitis.

\u2002 Retinoids have been used for the treatment of skin disorders such as acne, , and photoaging. However, despite their beneficial effects, topical retinoids often cause severe local irritation called retinoid dermatitis. We previously developed a novel vitamin A derivative, retinyl retinoate, which induces less irritation and affords excellent tolerance. In this study, we examined whether co-treatment with topical peroxisome proliferator-activated receptor-α (PPARα) agonists (e.g. WY14643) reduce retinoid dermatitis in hairless mouse skin.\u2002 The effect of concomitant treatment with a PPARα agonist on retinoid dermatitis in hairless mouse epidermis was evaluated by measuring transepidermal water loss, epidermal histology, and cytokine expression.\u2002 Retinyl retinoate induced less severe retinoid dermatitis than retinoic . Topical application of a PPARα agonist improved the stratum corneum structure and function, reduced mRNA expression of interleukin (IL)-1α, tumor necrosis factor-α and IL-8, and inhibited ear edema induced by retinoic or retinyl retinoate.\u2002 Our results indicate that PPARα agonists can potentially be used to improve retinoid dermatitis. We suggest that co-treatment with retinyl retinoate and a PPARα agonist may reduce or prevent detrimental alterations in retinoid-treated skin.© 2012 The International Society of Dermatology.

Keyword: psoriasis

Obesity exacerbates imiquimod-induced -like epidermal hyperplasia and interleukin-17 and interleukin-22 production in mice.

is a chronic inflammatory skin disorder that is accompanied by an imbalance between the proliferation and differentiation of keratinocytes. A number of studies have suggested an association between obesity and severe ; however, it remains to be clarified whether obesity exacerbates . To address this unsolved question, we induced psoriasiform dermatitis in mouse models for obesity. We found that obesity exaggerated the severity of psoriasiform dermatitis induced by topical application of the Toll-like receptor (TLR) 7 agonist, imiquimod. Ear swelling and epidermal hyperplasia were more prominent in the obese mice than in the control mice. When compared to imiquimod-treated control mice, imiquimod-treated obese mice expressed higher levels of mediators, interleukin-17A (IL-17A) and IL-22 in the skin. Food intake restriction partially abrogated enhanced ear swelling and cytokine overproduction in obese mice. Furthermore, the obesity environment and imiquimod treatment synergistically induced an IL-17A downstream molecule, regenerating islet-derived 3γ (Reg3γ), which is a critical molecule for psoriatic epidermal hyperplasia. , one of the fatty acids released by subcutaneous adipocytes, increased the expression of REG3A (a human homologue of mouse Reg3γ) in both the HaCaT keratinocyte cell line and normal human keratinocytes. Taken together, these results strongly suggest that obesity exacerbates psoriasiform dermatitis in mice by upregulating IL-17A, IL-22 and Reg3γ.© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: psoriasis

Effects of a mixture of fatty acids from sugar cane (Saccharum officinarum L.) wax oil in two models of inflammation: zymosan-induced arthritis and mice tail test of .

A mixture of fatty acids obtained from sugar cane (Saccharum officinarum L.) wax oil (FAM), in which the main constituents are , oleic, linoleic, and linolenic acids, was evaluated in two models of inflammation: zymosan-induced arthritis and in the tail test for , both on mice. In the first model, FAM significantly reduced zymozan-induced increase of beta glucuronidase (DE(50) 90+/-7 mg/kg). Histopathological studies showed inhibition in cellular infiltration and reduction of synovial hyperplasia and synovitis, whereas in the second test, histopathological and ultrastructural studies showed that topical application of FAM induced orthokeratosis with the presence of keratohyalin granules in the previously parakeratotic adult mouse tail, and without effects on epidermal thickness. The ED(50) of FAM in this model was 155+/-10 mg. The results of our studies showed that topical application of FAM exerts an important anti-inflammatory activity in both tests without evidence of irritant effects. The anti-inflamatory effects exerted by FAM may be due to its inhibitory effects on arachidonic metabolism. To our knowledge, this is the first report on the anti-inflammatory effect of sugar cane by-products in experimental models of arthritis and .

Keyword: psoriasis

Exacerbation and Prolongation of Psoriasiform Inflammation in Diabetic Obese Mice: A Synergistic Role of CXCL5 and Endoplasmic Reticulum Stress.

Accumulating evidence suggests that is frequently accompanied by metabolic disorders, such as obesity and diabetes. However, the mechanisms underlying the association between increased severity and concomitant metabolic syndrome have not been fully clarified. Herein, we show that imiquimod-induced psoriasiform inflammation was exacerbated and prolonged in diabetic obese mice compared to that in control mice, accompanied by remarkably increased lesional expressions of Cxcl5 and Il-1b. Notably, a large number of CXCL5 Ly6G cells infiltrated the dermis and subcutaneous fat tissue of the diabetic obese mice. Most macrophages in the subcutaneous fat tissues of the diabetic obese mice were positive for expression of IL-1β and GRP78/Bip, an endoplasmic reticulum stress marker. Depletion of Ly6G cells and macrophages diminished the imiquimod-induced psoriasiform inflammation. Further, CXCL5 potentiated the secretion of IL-1β from macrophages and , a fatty released from subcutaneous adipocytes, further enhanced IL-1β secretion via endoplasmic reticulum stress induction. Combined with the fact that the serum levels of both CXCL5 and are significantly elevated in patients with metabolic syndrome, our results suggest a role for CXCL5 and endoplasmic reticulum stress in the increase of severity of patients with concomitant metabolic syndrome.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: psoriasis

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P38 MAPK Pharmacological Inhibitor SB203580 Alleviates Total Parenteral Nutrition-Induced Loss of Intestinal but Promotes Hepatocyte Lipoapoptosis.

Our previous studies have provided evidence that p38 mitogen-activated protein kinase (MAPK) is involved in total parenteral nutrition (TPN)-associated complications, but its exact effects and mechanisms have not been fully understood. This study aimed to evaluate the roles of p38 MAPK inhibitor SB203580 in the TPN-induced loss of intestinal and liver disease.A rodent model of TPN was used to analyze the roles of SB203580 in TPN-associated complications.Intestinal was evaluated by transepithelial electrical resistance (TER) and paracellular permeability in Caco-2 cells. The (PA) was used to induce hepatic lipoapoptosis in vitro. The lipoapoptosis was detected using Caspase-3/7 and lipid staining.In the present study, we showed that SB203580 treatment significantly suppressed TPN-mediated intestinal permeability in rats. SB203580 treatment significantly inhibited IL-1β-induced an increase in tight junction permeability of Caco-2 cells via repressing the p38/ATF-2 signaling. Unexpectedly, SB203580 treatment enhanced hepatic lipoapoptosis in the model of TPN. (PA)-induced hepatic lipoapoptosis in human liver cells was significantly augmented by the SB203580 treatment.We demonstrate that the p38 MAPK inhibitor SB203508 ameliorates intestinal but promotes hepatic lipoapoptosis in model of TPN.© 2017 The Author(s)Published by S. Karger AG, Basel.

Keyword: tight junction

Adelmidrol + sodium hyaluronate in IC/BPS or conditions associated to chronic urothelial inflammation. A translational study.

Interstitial cystitis/painful bladder syndrome (IC/PBS) is a chronic bladder condition characterized by frequent urination, bladder inflammation and pain. It is a particular challenging disease and a clear unmet medical need in terms of identifying new therapeutic strategies. The aim of study was to evaluate the anti-inflammatory effects of intravesical Vessilen (a new formulation of 2% adelmidrol (the diethanolamide derivative of azelaic ) + 0.1% sodium hyaluronate) administration in rodent models of IC/BPS and in IC/BPS patients or other bladder disorders. Acute and chronic animal models of cystitis were induced by a single or repetitive intraperitoneal injections of cyclophosphamide (CYP); patients with IC/BPS or with bladder pain syndrome associated with symptoms of the lower urinary tract treated once weekly by bladder instillation of Vessilen for 8 weeks. CYP instillation caused macroscopic and histological bladder alterations, inflammatory infiltrates, increased mast cell numbers, bladder pain, increased expression of nitrotyrosine, decreased expression of endothelial zonula occludens-1. Intravesical Vessilen® treatment was able to ameliorate CYP induced bladder inflammation and pain by inhibiting nuclear factor-κB pathway and inflammatory mediator levels as well as reduced mechanical allodynia and nerve growth factor levels. A significant improvement in quality of life and symptom intensity were evident in patients with IC/BPS or other bladder disorders treated with Vessilen. Vessilen could be a new therapeutic approach for human cystitis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: tight junction

-Palmitoylation of Junctional Adhesion Molecule C Regulates Its Tight Junction Localization and Cell Migration.

Junctional adhesion molecule C (JAM-C) is an immunoglobulin superfamily protein expressed in epithelial cells, endothelial cells, and leukocytes. JAM-C has been implicated in leukocyte transendothelial migration, angiogenesis, cell adhesion, cell polarity, spermatogenesis, and metastasis. Here, we show that JAM-C undergoes -palmitoylation on two juxtamembrane cysteine residues, Cys-264 and Cys-265. We have identified DHHC7 as a JAM-C palmitoylating enzyme by screening all known palmitoyltransferases (DHHCs). Ectopic expression of DHHC7, but not a DHHC7 catalytic mutant, enhances JAM-C -palmitoylation. Moreover, DHHC7 knockdown decreases the -palmitoylation level of JAM-C. Palmitoylation of JAM-C promotes its localization to tight junctions and inhibits transwell migration of A549 lung cancer cells. These results suggest that -palmitoylation of JAM-C can be potentially targeted to control cancer metastasis.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: tight junction

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: tight junction

From the Cover: Comparative Proteomics Reveals Silver Nanoparticles Alter Fatty Metabolism and Amyloid Beta Clearance for Neuronal Apoptosis in a Triple Cell Coculture Model of the Blood-Brain .

Silver nanoparticles (AgNPs) enter the central nervous system through the blood-brain (BBB). AgNP exposure can increase amyloid beta (Aβ) deposition in neuronal cells to potentially induce Alzheimer's disease (AD) progression. However, the mechanism through which AgNPs alter BBB permeability in endothelial cells and subsequently lead to AD progression remains unclear. This study investigated whether AgNPs disrupt the tight junction proteins of brain endothelial cells, and alter the proteomic metabolism of neuronal cells underlying AD progression in a triple cell coculture model constructed using mouse brain endothelial (bEnd.3) cells, mouse brain astrocytes (ALT), and mouse neuroblastoma neuro-2a (N2a) cells. The results showed that AgNPs accumulated in ALT and N2a cells because of the disruption of tight junction proteins, claudin-5 and ZO-1, in bEnd.3 cells. The proteomic profiling of N2a cells after AgNP exposure identified 298 differentially expressed proteins related to fatty metabolism. Particularly, AgNP-induced production was observed in N2a cells, which might promote Aβ generation. Moreover, AgNP exposure increased the protein expression of amyloid precursor protein (APP) and Aβ generation-related secretases, PSEN1, PSEN2, and β-site APP cleaving enzyme for APP cleavage in ALT and N2a cells, stimulated Aβ40 and Aβ42 secretion in the culture medium, and attenuated the gene expression of Aβ clearance-related receptors, P-gp and LRP-1, in bEnd.3 cells. Increased Aβ might further aggregate on the neuronal cell surface to enhance the secretion of inflammatory cytokines, MCP-1 and IL-6, thus inducing apoptosis in N2a cells. This study suggested that AgNP exposure might cause Aβ deposition and inflammation for subsequent neuronal cell apoptosis to potentially induce AD progression.© The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Keyword: tight junction

Establishment and characterisation of a novel bovine SV40 large T-antigen-transduced foetal hepatocyte-derived cell line.

Due to lack of in vitro models for bovine hepatocytes apart from primary cells, there is demand for a bovine hepatocyte-derived cell line. Transduction of bovine foetal hepatocytes with SV40 large T-antigen was performed using the vector pRetro-E2 SV40. Phase contrast microscopy was carried out to evaluate morphology. Immunofluorescence staining was conducted to study expression of keratins, tight junction proteins zona occludens-1 and claudin-1, glucose transporter-2 and P-glycoprotein as well as phosphoenolpyruvate carboxykinase. Urea and triglyceride production was quantified photometrically. Histochemical staining of glycogen by Periodic -Schiff stain and of lipids with Oil red O was performed after 24\xa0h incubation with 20\xa0mM glucose and 85\xa0μM , respectively. Gene expression analysis of hepatocyte-typical genes was conducted by reverse transcription PCR. We obtained a SV40LTAg-transduced extended passage cell line, referred to as BFH12. Polygonal growth, keratins, tight junction proteins zona occludens-1 and claudin-1 and glucose transporter-2 as well as P-glycoprotein and phosphoenolpyruvate carboxykinase were attested positively. Urea production calculated as cell-specific rate was 14.2\u2009±\u20092.0\xa0fmol/h (early passage) and 17.6\u2009±\u20093.7\xa0fmol/h (late passage). Cell-specific triglyceride production was 1.6\u2009±\u20090.5\xa0fmol/h (early passage) and 2.1\u2009±\u20090.3\xa0fmol/h (late passage). Additionally, cells were positive for glycogen and lipid storage and showed a gene expression pattern resembling foetal hepatocytes. With the properties described here, the novel cell line BFH12 is a hepatocyte-derived cell line which can be used as an in vitro whole cell model.

Keyword: tight junction

High-fat diet promotes experimental colitis by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental colitis in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental colitis model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe colitis compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes colitis by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes colitis in a 2,4,6-trinitrobenzenesulfonic -induced experimental colitis model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: tight junction

Time-Dependent alteration to the structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal microbiota composition, increased intestinal permeability, and onset of type 2 diabetes mellitus (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the microbiota, after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: tight junction

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Long-term supplementation interacts with parity in lactating dairy cows: Production responses, nutrient digestibility, and energy partitioning.

The objective of our study was to evaluate the effects of long-term (C16:0) supplementation and parity on production, nutrient digestibility, and energy partitioning of mid-lactation dairy cows. Forty mid-lactation Holstein cows (18 primiparous and 22 multiparous) were used in a block design. Cows were assigned to receive either a control diet containing no supplemental fat (CON) or a C16:0-enriched supplemented diet (PA; 1.5% diet dry matter) fed for 10 wk. Compared with CON, PA increased dry matter intake, milk yield, cumulative milk yield, milk fat content, milk fat yield, 16-carbon milk fatty (FA) yield, 3.5% fat-corrected milk yield, and energy-corrected milk yield. Additionally, PA increased change, but did not affect condition score change compared with CON. A tendency for a treatment by parity interaction was observed for milk yield due to PA increasing milk yield in multiparous but not in primiparous cows. In addition, we observed interactions between treatment and parity for fat-corrected milk, energy-corrected milk, and milk fat yield due to PA increasing these variables to a greater extent in multiparous compared with primiparous cows. Interestingly, we observed an interaction between treatment and parity for change, due to PA increasing change in primiparous but not in multiparous cows. The PA treatment increased dry matter and neutral detergent fiber digestibilities compared with CON. Although PA did not affect 18-carbon FA digestibility, compared with CON, PA decreased 16-carbon and total FA digestibilities and increased total FA intake by 470 g/d and absorbed total FA by 316 g/d. We also observed an interaction between treatment and parity for total absorbed FA due to PA increasing it to a greater extent in multiparous than in primiparous cows. Compared with CON, PA increased apparent energy intake and milk energy output. We observed an interaction between treatment and parity for milk energy output due to PA increasing milk energy output to a greater extent in multiparous than primiparous cows. Additionally, an interaction between treatment and parity was observed for energy output in reserves due to PA increasing energy output in reserves in primiparous but not in multiparous cows. In conclusion, production responses of dairy cows to PA were consistent throughout the 10-wk treatment period. In addition, PA supplementation interacted with parity, with production responses increased to a greater extent in multiparous than primiparous cows and energy partitioned to reserves only increased in primiparous cows.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: weight

Polymorphism in the Gene Is Associated to Changes in Fatty Metabolism and Circulating Endocannabinoid Levels Distinctively in Normal and Obese Subjects.

The multifunctional CD36 scavenger receptor facilitates fatty (FA) uptake and oxidation and it has been involved in the pathophysiology related to dysfunctional FA metabolism. The common variant in the gene, (A/G), whose allele A is characterized by a reduced protein expression, has been associated with taste sensitivity to and preference for fat. We therefore aimed at evaluating whether the polymorphism may influence fatty metabolism and endocannabinoid biosynthesis in normal (NW) and obese (OB) subjects. Red blood cell (RBC) fatty composition, and plasma endocannabinoid levels were determined. In NW subjects with AA genotype was found a marked reduction of RBC saturated fatty acids and /linoleic ratio (PA/LA), considered as lipogenesis (DNL) biomarkers. Remarkably, to the reduction of DNL biomarkers corresponded an increase of omega-6 index, an indirect marker of the impact on fatty metabolism of dietary omega-6 fatty acids, endocannabinoid levels and a higher waist/hip ratio. The presence of the G allele was instead associated with increased endocannabinoid plasma levels and a trend for increased waist/hip ratio in obese subjects, even though exhibited decreased BMI with respect to those with AA genotype. These data indicate that the polymorphism, , leads to a distinct metabolic pattern in NW and in OB subjects. Therefore, their determination may be crucial in developing personalized therapeutic strategies for ameliorating dyslipidemia and other metabolic disorders.

Keyword: weight

Assessing biodiesel quality parameters for wastewater grown Chlorella sp.

Microalgae are reported as the efficient source of renewable biodiesel which should be able to meet the global demand of transport fuels. Present study is focused on assessment of wastewater grown indigenous microalga Chlorella sp. for fuel quality parameters. This was successfully grown in secondary treated waste water diluted with tap water (25% dilution) in glass house. The microalga showed a dry of 0.849 g L with lipid content of 27.1% on dry basis on 21st day of incubation. After transesterification, the yield of fatty methyl ester was 80.64% with major fatty acids as , linoleic, oleic and linolenic. The physical parameters predicted from empirical equations in the biodiesel showed cetane number as 56.5, iodine value of 75.5 g I 100 g, high heating value 40.1 MJ kg, flash point 135 °C, kinematic viscosity 4.05 mm s with density of 0.86 g cm and cold filter plugging point as 0.7 °C. Fourier transform infra-red (FTIR), H, C NMR spectrum confirmed the chemical nature of biodiesel. The results indicated that the quality of biodiesel was almost as per the criterion of ASTM standards; hence, wastewater grown Chlorella sp. can be used as a promising strain for biodiesel production.

Keyword: weight

Trigonelline prevents high cholesterol and high fat diet induced hepatic lipid accumulation and lipo-toxicity in C57BL/6J mice, via restoration of hepatic autophagy.

Non-alcoholic fatty liver disease (NAFLD) is often linked with impaired hepatic autophagy. Here, we studied the alterations in hepatocellular autophagy by high cholesterol and high-fat diet (HC-HF) diet in C57BL/6J mice, and by (PA), in AML-12 and HepG2 cells. Further, we analysed role of Trigonelline (TG), a plant alkaloid, in preventing NAFLD, by modulating autophagy. For this, C57BL/6J mice were fed with Standard Chow (SC) or HC-HF diet, with and without TG for 16 weeks. In-vitro; AML-12\u202fcells and HepG2 cells, were exposed to PA with and without TG, for 24\u202fh. Cellular events related to autophagy, lipogenesis, and lipo-toxicity were studied. The HC-HF diet fed mice showed hepatic autophagy blockade, increased triglycerides and steatosis. PA exposure to AML-12\u202fcells and HepG2 cells induced impaired autophagy, ER stress, resulting in lipotoxicity. TG treatment in HC-HF fed mice, restored hepatic autophagy, and prevented steatosis. TG treated AML-12, and HepG2 cells exposed to PA showed autophagy restoration, and reduced lipotoxicity, however, these effects were diminished in Atg7-/- HepG2 cells, and in the presence of chloroquine. This study shows that HC-HF diet-induced impaired autophagy, and steatosis is prevented by TG, which attributes to its novel mechanism in treating NAFLD.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: weight

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to lipid metabolism and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: weight

Phenolics, tocopherols and fatty profiling of wild and commercial mushrooms from Pakistan.

Mushrooms can be used as nutraceutical or functional foods to maintain and promote good health. In the present study, wild Ganoderma lucidum and four commercial mushrooms, Pleurotus ostreatus, Volvariella volvacea, Hericium erinaceus and Lentinus edodes, collected from Pakistan were screened for phenolics, tocopherols and fatty contents. High performance liquid chromatography analysis of phenolic acids showed that chlorogenic , ferulic , gallic , p-Coumaric and caffeic acids were observed in selected mushrooms. H. erinaceus contained high amounts of chlorogenic (11.49±0.1 µ/g of dry ) and ferulic (7.84±0.7 µg/g of dry ). γ-tocopherol and lutein were present in all studied mushrooms. Lutein contents were higher in H. erinaceus (2.42±0.087 µg/g of DW) followed by V. volvacea> P. ostreatus> L. edodes. γ-tocopherol was observed in the range of 74.25±3.01 to 29.65±1.2 µg/g of dry . GC/MS analysis of fatty acids showed that linoleic (18\x92:2n6c), oleic (18\x92:1n9c), (C16:0), stearic (C18:0), linolenic (18\x92:3n3) and nonadecanoic (C19-0), were the main fatty acids found in selected mushrooms. The unsaturated fatty acids were predominated over saturated fatty acids. It is concluded that selected mushrooms are good sources of antioxidant compounds and unsaturated fatty acids.

Keyword: weight

Glucagon-like peptide-1 analog prevents obesity-related glomerulopathy by inhibiting excessive autophagy in podocytes.

To investigate the role of glucagon-like peptide-1 analog (GLP-1) in high-fat diet-induced obesity-related glomerulopathy (ORG). Male C57BL/6 mice fed a high-fat diet for 12 wk were treated with GLP-1 (200 μg/kg) or 0.9% saline for 4 wk. Fasting blood glucose and insulin and the expression of podocin, nephrin, phosphoinositide 3-kinase (PI3K), glucose transporter type (Glut4), and microtubule-associated protein 1A/1B-light chain 3 (LC3) were assayed. Glomerular morphology and podocyte foot structure were evaluated by periodic -Schiff staining and electron microscopy. Podocytes were treated with 150 nM GLP-1 and incubated with 400 μM (PA) for 12 h. The effect on autophagy was assessed by podocyte-specific Glut4 siRNA. Insulin resistance and autophagy were assayed by immunofluorescence and Western blotting. The high-fat diet resulted in gain, ectopic glomerular lipid accumulation, increased insulin resistance, and fusion of podophyte foot processes. The decreased translocation of Glut4 to the plasma membrane and excess autophagy seen in mice fed a high-fat diet and in PA-treated cultured podocytes were attenuated by GLP-1. Podocyte-specific Glut4 siRNA promoted autophagy, and rapamycin-enhanced autophagy worsened the podocyte injury caused by PA. Excess autophagy in podocytes was induced by inhibition of Glut4 translocation to the plasma membrane and was involved in the pathology of ORG. GLP-1 restored insulin sensitivity and ameliorated renal injury by decreasing the level of autophagy.

Keyword: weight

Evaluation of the Stability of Polymeric Materials Exposed to Palm Biodiesel and Biodiesel⁻Organic Blends.

The aim of the present work is to evaluate the impact of pure palm biodiesel fuel (B100) and biodiesel blends with 0.32% oleic, , acetic, myristic, and stearic acids on the properties of some polymeric materials used commonly in the manufacture of auto parts such as the polyamide 66 (PA66), polyoxymethylene (POM), and high-density polyethylene (HDPE). The effects of the B100 and B100⁻ blends on polymeric materials were examined by comparing changes in the gain/loss of mass and by measuring the hardness, the impact strength, and the tensile strength of the materials at the end of the exposure. The characterization of the polymers was carried out before and after exposure by using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and Fourier transform infrared spectroscopy (FTIR). After the immersion in B100⁻acids blends, the HDPE exhibited an increase in mass of 5%, which was very similar in all blends. The PA66 showed a small decrease in (2% approx.) in all mixtures. The POM presented an increase in the percentage of in the mixture of B100 with acetic of 0.3%. A decrease was observed in the crystallinity of the HDPE when exposed to blends of B100⁻acids. This behavior may be associated with a plasticizing effect in the HDPE exposed to the blends. The mechanical properties of POM and HDPE showed no significant changes after immersion in the fuels. On the other hand, PA66 exhibited a significant decrease in maximum stress value after immersion in B100, B100⁻oleic and B100⁻ blends. The variation of the mechanical properties of the PA66 after exposure to B100 was potentiated by addition of organic acids. The assessed polymers did not undergo appreciable changes in the chemical structure of the samples after immersion in the fuels, so the variation in the mechanical properties could be explained by physical absorption of the fuel into the polymers.

Keyword: weight

GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic fatty homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced liver dysfunction, hepatic steatosis and insulin resistance. In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and lipogenesis, and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), fatty synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting lipogenesis and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the liver may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: weight

Levels of ester of hydroxystearic (PAHSA) are reduced in the breast milk of obese mothers.

To achieve optimal development of a newborn, breastfeeding is extensively recommended, but little is known about the role of non-nutritive bioactive milk components. We aimed to characterize the fatty esters of hydroxy fatty acids (FAHFAs), namely hydroxystearic acids (PAHSAs)-endogenous lipids with anti-inflammatory and anti-diabetic properties, in human breast milk. Breast milk samples from 30 lean (BMI=19-23) and 23 obese (BMI>30) women were collected 72h postpartum. Adipose tissue and milk samples were harvested from C57BL/6J mice. FAHFA lipid profiles were measured using reverse phase and chiral liquid chromatography-mass spectrometry method. PAHSA regioisomers as well as other FAHFAs were present in both human and murine milk. Unexpectedly, the levels of 5-PAHSA were higher relative to other regioisomers. The separation of both regioisomers and enantiomers of PAHSAs revealed that both R- and S-enantiomers were present in the biological samples, and that the majority of the 5-PAHSA signal is of R configuration. Total PAHSA levels were positively associated with gain during pregnancy, and 5-PAHSA as well as total PAHSA levels were significantly lower in the milk of the obese compared to the lean mothers. Our results document for the first time the presence of lipid mediators from the FAHFA family in breast milk, while giving an insight into the stereochemistry of PAHSAs. They also indicate the negative effect of obesity on 5-PAHSA levels. Future studies will be needed to explore the role and mechanism of action of FAHFAs in breast milk.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: weight

Estimates of genetic parameters for fatty compositions in the longissimus dorsi muscle of Hanwoo cattle.

We estimated the heritabilities (h 2) and genetic and phenotypic correlations among individual and groups of fatty acids, as well as their correlations with six important carcass and meat-quality traits in Korean Hanwoo cattle. Meat samples were collected from the longissimus dorsi muscles of 1000 Hanwoo steers that were 30-month-old (progeny of 85 proven Hanwoo bulls) to determine intramuscular fatty profiles. Phenotypic data on carcass (CWT), eye muscle area (EMA), back fat thickness (BFT), marbling score (MS), Warner-Bratzler shear force (WBSF) and intramuscular fat content (IMF) were also investigated using this half-sib population. Variance and covari.ance components were estimated using restricted maximum likelihood procedures under univariate and pairwise bivariate animal models. Oleic (C18:1n-9) was the most abundant fatty , accounting for 50.69% of all investigated fatty acids, followed by (C16:0; 27.33%) and stearic (C18:0; 10.96%). The contents of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were 41.64%, 56.24% and 2.10%, respectively, and the MUFA/SFA ratio, PUFA/SFA ratio, desaturation index (DI) and elongation index (EI) were 1.36, 0.05, 0.59 and 0.66, respectively. The h 2 estimates for individual fatty acids ranged from very low to high (0.03±0.14 to 0.63±0.14). The h 2 estimates for SFAs, MUFAs, PUFAs, DI and EI were 0.53±0.14, 0.49±0.14, 0.23±0.10, 0.51±0.13 and 0.53±0.13, respectively. The genetic and phenotypic correlations among individual fatty acids and fatty classes varied widely (-0.99 to 0.99). Notably, C18:1n-9 had favourable (negative) genetic correlations with two detrimental fatty acids, C14:0 (-0.76) and C16:0 (-0.92). Genetic correlations of individual and group fatty acids with CWT, EMA, BFT, MS, WBSF and IMF ranged from low to moderate (both positive and negative) with the exception of low-concentration PUFAs. Low or near-zero phenotypic correlations reflected potential non-genetic contributions. This study provides insights on genetic variability and correlations among intramuscular fatty acids as well as correlations between fatty acids and carcass and meat-quality traits, which could be used in Hanwoo breeding programmes to improve fatty compositions in meat.

Keyword: weight

Does different BMI influence oocyte and embryo quality by inducing fatty in follicular fluid?

We aimed to assess the effects of obesity-related follicular fluid (FF) fatty acids (FAs) on the number and quality of oocytes, good embryo quality rate, and pregnancy rate.This prospective cohort study was conducted on 105 infertile women under the age of 38, who underwent intracytoplasmic sperm injection (ICSI) from March 2015 to October 2015. They were grouped into three mass index (BMI) categories. The fatty acids composition of the FF was analyzed by GC-MS head space method. We studied the FAs correlation with BMI and ICSI outcomes.The distribution of fatty acids did not differ significantly in each BMI group, with the exception for stearic that was marginally significant (p=0.05). The mean number of mature oocytes did not differ significantly between the BMI groups, the percent of Metaphase II (MII) oocytes was inversely associated with the BMI (r=-0.21, p=0.03). Kruskal-Wallis test showed that the distribution of good quality embryos\' percentages were different in at least two categories of studied BMI groups (p=0.009, p=0.02). The mean concentration of was higher in nonpregnant patients for all of the studied BMI classes (p=0.02, p=0.03, p=0.05), however, stearic (p<0.001) and linolenic acids (p=0.01) were higher in nonpregnant normal patients.Differences in BMI are not associated with the fatty composition of the FF. The FF fatty possibly affects the outcome of ICSI through the achievement of clinical pregnancy. Therefore, it is essential to provide patients with nutritional counseling before they use assisted reproductive techniques.Copyright © 2017. Published by Elsevier B.V.

Keyword: weight

Optimization of enzymatic hydrolysis of cellulosic fraction obtained from stranded driftwood feedstocks for lipid production by .

Stranded driftwood feedstocks may represent, after pretreatment with steam explosion and enzymatic hydrolysis, a cheap C-source for producing biochemicals and biofuels using oleaginous yeasts. The hydrolysis was optimized using a response surface methodology (RSM). The solid loading (SL) and the dosage of enzyme cocktail (ED) were variated following a central composite design (CCD) aimed at optimizing the conversion of carbohydrates into lipids (Y) by the yeast DBVPG 5870. A second-order polynomial equation was computed for describing the effect of ED and SL on Y. The best combination (ED\u202f=\u202f3.10%; SL\u202f=\u202f22.07%) for releasing the optimal concentration of carbohydrates which gave the highest predicted Y (27.32%) was then validated by a new hydrolysis. The resulting value of Y (25.26%) was close to the theoretical maximum value. Interestingly, fatty profile achieved under the optimized conditions was similar to that reported for palm oil.

Keyword: weight

Comparative compositional analysis of transgenic potato resistant to potato tuber moth (PTM) and its non-transformed counterpart.

In this study, the compositions of transgenic potatoes (TPs) resistant to potato tuber moth (Phthorimaea operculella) were compared with those of its non-transgenic (NTP) counterparts. The light inducible promoter, phosphoenolpyruvate carboxylase led to the expression of Cry1Ab only in the leaves and light-treated tubers of the TPs. No significant differences were found in the moisture, ash, dry , total soluble protein, carbohydrate, starch, fiber, ascorbate, cations, anions, fatty acids, and glycoalkaloids contents of TP and NTP. Moreover, light treatment significantly affected the contents of ascorbate, acetate and nitrite anions, , stearic and linolenic fatty acids, α-haconine and α-solanine glycoalkaloids in TP and NTP tubers. While, significant differences were observed in the amino contents in light-treated tubers of TPs than the NTP ones. Although, light treatment in potato tubers resulted in marked metabolic changes, all the variations observed in the metabolites compositions were found to be within the desired reference ranges for potato plants. In conclusion, the results indicated that the TPs were substantially and nutritionally equivalent to the NTP counterparts.

Keyword: weight

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell and the lipid yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: weight

Good Fats versus Bad Fats: A Comparison of Fatty Acids in the Promotion of Insulin Resistance, Inflammation, and Obesity.

Recently, debate has erupted in both the scientific community and throughout the lay public around whether a low-fat or low-carbohydrate diet is better for loss. In other words, is it better to cut fat or cut carbohydrate for loss. However, going beyond this debate (fat versus carbohydrate), are questions around whether certain fatty acids are worse for promoting insulin resistance, inflammation, and obesity. The overall evidence in the literature suggests that medium-chain saturated fats (such as lauric , found in coconut oil) and monounsaturated fat (oleic , found in olive oil) are less likely to promote insulin resistance, inflammation, and fat storage compared to long-chain saturated fatty acids (such as stearic found in large quantities in butter, but particularly found in palm oil) especially when consumed on top of a diet moderate in refined carbohydrates. Compared to long-chain saturated fats, lauric and oleic have an increased fatty oxidation rate, are more likely to be burned for energy and less likely to be stored in adipose tissue, and thus promote increased energy expenditure. Omega-6 polyunsaturated fatty acids (PUFAs), such as linoleic , as found in vegetable oils may contribute to obesity, whereas omega-3 PUFA may be protective. Importantly, both olive oil as part of a Mediterranean diet, and omega-3 from fish and fish oil have been proven to reduce risk of cardiovascular (CV) events.

Keyword: weight

Chemical and Nutritional Composition of Terminalia ferdinandiana (Kakadu Plum) Kernels: A Novel Nutrition Source.

(Kakadu plum) is a native Australian fruit. Industrial processing of fruits into puree generates seeds as a by-product, which are generally discarded. The aim of our present study was to process the seed to separate the kernel and determine its nutritional composition. The proximate, mineral and fatty compositions were analysed in this study. Kernels are composed of 35% fat, while proteins account for 32% dry (DW). The energy content and fiber were 2065 KJ/100 g and 21.2% DW, respectively. Furthermore, the study showed that kernels were a very rich source of minerals and trace elements, such as potassium (6693 mg/kg), calcium (5385 mg/kg), iron (61 mg/kg) and zinc (60 mg/kg) DW, and had low levels of heavy metals. The fatty composition of the kernels consisted of omega-6 fatty , linoleic (50.2%), monounsaturated oleic (29.3%) and two saturated fatty acids namely (12.0%) and stearic (7.2%). The results indicate that kernels have the potential to be utilized as a novel protein source for dietary purposes and non-conventional supply of linoleic, and oleic acids.

Keyword: weight

Fabrication and Performance of Composite Microencapsulated Phase Change Materials with Ethyl Ester as Core.

Microencapsulation of phase change materials (PCMs) could prevent the leakage of PCMs during solid⁻liquid phase change process. However, their applications are mainly limited by the compactness and thermal stability of the traditional polyurea shell microcapsules. To increase the thermal compactness and thermal stability of PCM microcapsules, tetraethylorthosilicate (TEOS) was employed to form polymer/SiO₂ composite shells to enhance the mechanical performance of polyurea and polyurethane microcapsule via interfacial polymerization and in situ polymerization. The morphology and chemical components of the microcapsules were characterized by field-emission scanning electron microscope (FE-SEM) and Fourier transform infrared (FT-IR) spectroscopy, respectively. The thermal properties of the microcapsules were investigated by differential scanning calorimetry (DSC) and thermal gravity analysis (TGA). The results showed the smoothness and compactness of both polyurea⁻SiO₂ and polyurethane⁻SiO₂ microcapsules enhanced slightly, when compared with that without TEOS addition. Moreover, the SiO₂ composite shell had good effect on thermal compactness, as the loss rate of polyurea⁻SiO₂ microcapsules and polyurethane⁻SiO₂ microcapsules decreased 3.5% and 4.1%, respectively.

Keyword: weight

Chemical composition and biological activity of the essential oil from the root of Courb. native to Saudi Arabia.

The chemical composition of the essential oil from roots was determined via GC-FID. There were 80 compounds, representing 99.99% of the total oil constituents. Among these, 77.31% were sesquiterpenes, 14.62% were fatty acids, 7.21% were other components (i.e., phenolics, hydrocarbons, etc.), and 0.85% were monoterpenes. The major compounds in the oil were γ-eudesmol (35.31%), 5-guaien-11-ol (14.43%), -cedrol (8.19%), oleic (5.23%), bulnesol (4.45%), α-linoleic (4.20%), 3,4-dimethoxycinnamic (3.83%), (2.69%), isolongifolanone (2.68%), eicosane (1.41%), and cedrol (1.14%). Oxygenated sesquiterpenes were found to represent more than 50% percent of the total oil content. Moreover, the essential oil was evaluated for anti-inflammatory, antioxidant, antipyretic, and antinociceptive activities using and models. Additionally, the antioxidant potential of the oil was evaluated using various\xa0 antioxidant tests, including DPPH, ABTS and FRAP. At a dose of 240\u202fµl/kg, the oil showed anti-inflammatory (59.12%), antipyretic (37.00\u202f±\u202f0.11), and antinociceptive (47.58%) activities and showed significant (p\u202f<\u202f0.001) effect as compared to a standard drug (phenylbutazone and indomethacin). These findings demonstrated that the essential oil of root could be used as a natural source for their anti-inflammatory, antinociceptive, antipyretic, and antioxidant effects.

Keyword: weight

Fasting rapidly increases fatty oxidation in white adipose tissue of young broiler chickens.

Upregulating the fatty oxidation capacity of white adipose tissue in mice protects against diet-induced obesity, inflammation and insulin resistance. Part of this capacity results from induction of brown-like adipocytes within classical white depots, making it difficult to determine the oxidative contribution of the more abundant white adipocytes. Avian genomes lack a gene for uncoupling protein 1 and are devoid of brown adipose cells, making them a useful model in which to study white adipocyte metabolism in vivo. We recently reported that a brief (5\xa0hour) period of fasting significantly upregulated many genes involved in mitochondrial and peroxisomal fatty oxidation pathways in white adipose tissue of young broiler chickens. The objective of this study was to determine if the effects on gene expression manifested in increased rates of fatty oxidation. Abdominal adipose tissue was collected from 21\xa0day-old broiler chicks that were fasted for 3, 5 or 7\xa0hours or fed ad libitum (controls). Fatty oxidation was determined by measuring and summing CO production and C-labeled -soluble metabolites from the oxidation of [1-C] . Fasting induced a progressive increase in complete fatty oxidation and citrate synthase activity relative to controls. These results confirm that fatty oxidation in white adipose tissue is dynamically controlled by nutritional status. Identifying the underlying mechanism may provide new therapeutic targets through which to increase fatty oxidation in situ and protect against the detrimental effects of excess free fatty acids on adipocyte insulin sensitivity.

Keyword: weight

Effects of saturated and omega-3 polyunsaturated fatty acids on Sertoli cell apoptosis.

Obesity is believed to negatively affect male semen quality and is accompanied by dysregulation of free fatty (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report obesity decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive oxygen species were elevated. Furthermore, we demonstrated by in vitro assays that saturated (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated fatty acids might be the cause of obesity-induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs.FFA: free fatty ; HFD: high-fat diet; SD: standard diet; PA: ; PUFA: polyunsaturated fatty ; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive oxygen species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic fatty liver disease; DCFH-DA: 2\', 7\' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic ; PI: propidium iodide; DHA: docosahexenoic .

Keyword: weight

Dehydration Causes Increased Reliance on Protein Oxidation in Mice: A Test of the Protein-for-Water Hypothesis in a Mammal.

During fasting, animals rely on a mixture of fats, carbohydrates, and proteins that are derived solely from endogenous sources. The relative contributions of these metabolic fuels chiefly depend on the duration of the fast, but other factors including previous diet, environmental temperature, and activity level can modulate the fuel mixture. It has long been held that endogenous proteins are spared from catabolism until the final stages of prolonged fasting and contribute a significant proportion of energy once the other metabolic fuels have been depleted. However, evidence is mounting that protein is catabolized supplemental to fat metabolism under some circumstances. This has been shown in migratory birds that exhibit dramatic reductions in lean mass during flights. One hypothesis to explain this seemingly maladaptive metabolic strategy is that the catabolism and oxidation of protein in situ yields five times more metabolic water than that generated through fat oxidation alone. Recent support for this hypothesis is that birds benefit from such a strategy due to their uricotelic nature. However, it remains unclear whether ureotelic mammals would also employ this strategy. Here we test the protein-for-water hypothesis in resting laboratory mice subjected to water deprivation during fasting while we tracked rates of protein and lipid catabolism using endogenously incorporated C-leucine and C-. We found no differences in instantaneous leucine oxidation; however, cumulative differences in instantaneous leucine oxidation ultimately resulted in a higher total leucine oxidation after 72 h of fasting in water-deprived animals. We also found that lipid oxidation was 8% higher in the hydrated mice, but the difference was not significant presumably because of a concomitant reduction in metabolic rates of the water-deprived mice. Our results indicate that mammals do increase rates of protein catabolism during dehydration but to a lesser degree than birds. The ability of mammals to produce highly concentrated urine and their lower inherent rates of protein turnover apparently preclude mammals from taking full advantage of the protein-for-water strategy during fasting under dehydrating conditions.

Keyword: weight

Altering the ratio of dietary C16:0 and cis-9 C18:1 interacts with production level in dairy cows: Effects on production responses and energy partitioning.

The objective of our study was to evaluate the effects of altering the dietary ratio of (C16:0) and oleic (cis-9 C18:1) acids on nutrient digestibility, energy partitioning, and production responses of lactating dairy cows. Cows were blocked by milk yield and assigned to 3 groups (12 cows per group) in a main plot: low (45.2 ± 1.7 kg/d), medium (53.0 ± 1.6 kg/d), and high (60.0 ± 1.9 kg/d). Within each production group, a truncated Latin square arrangement of fatty (FA) treatments was used in 2 consecutive 35-d periods. The FA treatments supplemented at 1.5% of diet dry matter were (1) 80:10 (80% C16:0 + 10% cis-9 C18:1), (2) 73:17 (73% C16:0 + 17% cis-9 C18:1), (3) 66:24 (66% C16:0 + 24% cis-9 C18:1), and (4) 60:30 (60% C16:0 + 30% cis-9 C18:1). Treatment × production group interactions were observed for yields of milk, fat-corrected milk, energy-corrected milk, milk fat, milk protein, and milk lactose and energy partitioned to milk. Increasing cis-9 C18:1 in FA treatments reduced fat-corrected milk, energy-corrected milk, and milk energy output in low-producing cows but increased these in high-producing cows. Increasing cis-9 C18:1 in FA treatments did not affect milk yield, milk protein yield, and milk lactose yield in low- and medium-producing cows but increased these in high-producing cows. Regardless of production level, there was no effect of treatments on dry matter intake; however, increasing cis-9 C18:1 in FA treatments increased change and condition score change. Increasing cis-9 C18:1 in FA treatments increased total FA digestibility due to a linear increase in 16- and 18-carbon FA digestibilities. Interactions between FA treatments and production level were observed for the yield of milk fat and milk FA sources. In low-producing cows, increasing cis-9 C18:1 in FA treatments decreased milk fat yield due to a decrease in de novo and mixed milk FA without changes in preformed milk FA. In contrast, in high-producing cows, increasing cis-9 C18:1 in FA treatments increased milk fat yield due to an increase in de novo and preformed milk FA. Our results indicate that high-producing dairy cows (averaging 60 kg/d) responded better to a fat supplement containing more cis-9 C18:1, whereas low-producing cows (averaging 45 kg/d) responded better to a supplement containing more C16:0.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: weight

9-PAHSA promotes browning of white fat via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract obesity in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white fat. 9-PAHSA, an endogenous mammalian lipid, which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown fat specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown fat markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of obesity.Copyright © 2018. Published by Elsevier Inc.

Keyword: weight

Qualitative and quantitative analysis of lipo-alkaloids and fatty acids in Aconitum carmichaelii using LC-MS and GC-MS.

Lipo-alkaloid (LA) is a kind of C -norditerpenoid alkaloid in Aconitum species, which usually contains an aconitane skeleton and one or two fatty residues.To qualify and quantify the fatty acids and lipo-alkaloids in Aconitum carmichaelii.An ultra-high performance liquid chromatography-triple quadrupole-mass spectrometry (UHPLC-QQQ-MS) method was established to quantify LAs, while the free fatty acids were identified by gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF-MS).Six major LAs (1-6) containing linoleic, , and oleic residues as side chains were quantified. Eighteen fatty acids were determined by GC-MS, and 15 were detected as the side chains of LAs. The LAs containing these 15 fatty residues accounted for about a third of the total identified LAs. Moreover, the contents of linoleic, , and oleic acids were highest. In addition, 12 oxygenated fatty acids were also identified by UHPLC-Q-TOF-MS for the first time.The positive correlation between free fatty acids and LAs in A. carmichaelii indicated that the types and contents of LAs were influenced by free fatty acids.Copyright © 2018 John Wiley & Sons, Ltd.

Keyword: weight

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut microbiota and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: weight

CTRP3 attenuates high-fat diet-induced male reproductive dysfunction in mice.

Recent studies have suggested a role for abdominal obesity in male infertility. Previous studies have found that cell apoptosis exerts an important role in obesity-related male infertility. C1q/TNF-related protein 3 (CTRP3), a paralog of adiponectin, has been proposed to exert anti-apoptotic effects and to attenuate diabetes-related cardiac injuries. However, the role of CTRP3 in high-fat diet (HFD)-induced spermatogenic impairment remains unclear. In the present study, we fed male mice an HFD for 24 weeks to induce obesity. The expression of CTRP3 was decreased by HFD feeding. Supplementation with the recombinant human globular domain of CTRP3 (0.25 μg/g/day) for 4 weeks beginning at 20 weeks of the HFD improved spermatogenic function in the HFD-fed mice, which were characterized by improved testis morphology, increased testis / ratio, and increased sperm count, sperm viability, and sperm motility. We also found that CTRP3 infusion resulted in the attenuation of endoplasmic reticulum (ER) stress and the activation of silence information regulator 1 (SIRT1) in the testes of obese mice. Our study also suggested that CTRP3 attenuated the (PA)-induced reductions in sperm viability and motility via the inhibition of ER stress. Moreover, germ cell-specific knockout abolished the protective effects of CTRP3 and studies of human sperm showed that the protective effects of CTRP3 on sperm viability and motility were abrogated by a specific inhibitor of SIRT1. Thus, our results demonstrated that CTRP3 expression protected against HFD-induced spermatogenic deficiency through the SIRT1/ER stress pathway.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: weight

Short communication: Comparison of a -enriched triglyceride supplement and calcium salts of palm fatty acids supplement on production responses of dairy cows.

The objective of our study was to evaluate the effects of feeding a -enriched triglyceride supplement or a calcium salts of palm fatty (FA) supplement on nutrient digestibility and production responses of mid-lactation dairy cows. Fifteen Holstein cows (139 ± 39 d in milk) were randomly assigned to treatment sequence in a 3 × 3 Latin square design. Treatments were a control diet (CON; no fat supplement) and 1.5% of FA added either as a -enriched triglyceride supplement (PA-TG) or as calcium salts of palm FA supplement (Ca-FA). Fat-supplemented treatments did not affect dry matter intake (DMI) compared with CON, but Ca-FA reduced DMI compared with PA-TG. Compared with CON, fat-supplemented treatments increased 18-carbon FA digestibility by 2.0 percentage units but did not affect digestibility of total FA or 16-carbon FA. Compared with Ca-FA, PA-TG reduced total FA digestibility by 8.7 percentage units due to a decrease in 16-carbon FA digestibility (21.7 percentage units). Both fat supplements increased neutral detergent fiber (NDF) digestibility compared with CON (3.90 percentage units), and PA-TG tended to increase NDF digestibility by 1.60 percentage units compared with Ca-FA. Compared with CON, fat-supplemented treatments increased milk yield (1.05 kg/d), 3.5% fat-corrected milk yield (2.20 kg/d), and energy-corrected milk yield (1.80 kg/d). Also, PA-TG increased milk fat yield (50 g/d) and milk energy output (1.0 Mcal/d) and tended to increase milk fat content (0.07 percentage units) and energy-corrected milk yield (1.0 kg/d) compared with Ca-FA. Fat-supplemented treatments reduced the yield of de novo milk FA (23 g/d) and increased the yields of mixed (43 g/d) and preformed (52 g/d) milk FA compared with CON. The PA-TG treatment increased the yield of 16-carbon (66 g/d) milk FA compared with Ca-FA, whereas Ca-FA increased the yield of preformed (60 g/d) milk FA. Fat-supplemented treatments increased intake of net energy for lactation by 1.80 Mcal/d, milk energy output by 1.30 Mcal/d, and energy in reserves by 0.30 Mcal/d compared with CON. The Ca-FA treatment increased energy allocated to reserves (0.60 Mcal/d), energy partitioning toward reserves (1.20 percentage units), and condition score change (0.06 units), and tended to increase change (0.16 kg/d) and condition score (0.08 units) compared with PA-TG. In conclusion, feeding a -enriched triglyceride supplement increased milk energy output due to increased yields of milk and milk fat, whereas feeding a calcium salts of palm FA supplement increased FA digestibility and energy partitioned to reserves.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: weight

In-vitro and in-vivo studies supporting the therapeutic potential of ZP3022 in diabetes.

GLP-1-gastrin dual agonist ZP3022 has been shown to increase β-cell mass with a concomitant improvement of glycemic control in diabetic mice and rats. Here we tested the in-vitro effects of ZP3022 on β-cell proliferation, islet apoptosis and glucose-stimulated insulin secretion (GSIS) in rat islets of Langerhans. Moreover, gene expression profiling in whole pancreas from Zucker Diabetic Fatty (ZDF) rats was performed to characterize genes differently regulated by short-term treatment with ZP3022. Treatments with exendin-4, gastrin-17 alone or in combination were included in the studies. ZP3022 promoted β-cell proliferation, protected from palmitate-, but not from cytokine-induced apoptosis, and induced an increase in GSIS, demonstrating a glucose dependent insulinotropic action of ZP3022 on β-cells. The combination treatment with exendin-4 and gastrin-17 showed comparable effects on proliferation, apoptosis, and GSIS as did ZP3022. Microarray analysis revealed that ZP3022 exerted specific effects on pancreatic gene expression not observed when treating ZDF rats with either exendin-4 alone or in combination with gastrin-17. In particular MAPK signaling pathway was observed among the highest affected pathways; while also pathways related to insulin signaling and secretion were regulated by ZP3022. Moreover, rats treated with ZP3022 had a higher expression of genes encoding for the specific β-cell/endocrine cell markers, such as islet amyloid polypeptide (IAPP), protein convertase 1/3 and -2 (PC 1/3 and-2), as well as transmembrane protein 27(TMEM27) compared to vehicle treated rats. We conclude that ZP3022 may have therapeutic potential in the prevention/delay of β cell dysfunction.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: weight

Oriented surface epitope imprinted polymer-based quartz crystal microbalance sensor for cytochrome c.

A quartz crystal microbalance (QCM) sensor for detecting cytochrome c based on an oriented surface epitope imprinted polymer was fabricated in this paper. By using the -modified epitope of cytochrome c as the template and the 3-aminopropyltriethoxysilane as the monomer, we prepared a new oriented surface epitope imprinted polymer by the reverse microemulsion polymerization. The prepared oriented imprinted polymer had better imprinting effect than the non-oriented imprinted polymer. And compared to previous studies, this polymerization method is simple and could be carried out at room temperature in the presence of oxygen, under regular atmospheric conditions. Then, by combining the advantages of molecularly imprinted polymers and QCM sensors, we used the prepared polymer to establish a QCM sensor. The described sensor showed good sensitivity and selectivity towards cytochrome c. The linear range was from 0.005\u202fμg\u202fmL to 0.050\u202fμg\u202fmL and the detection limit was 3.6\u202fng\u202fmL which is lower than most of previous works. Besides, it could be used for real sample analysis and had satisfactory reproducibility and accuracy. This work proposed a new way of fabricating oriented surface epitope imprinted polymers-based QCM sensors for selectively detecting proteins at very low concentrations.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: weight

Cooperative stimulation of atherogenesis by lipopolysaccharide and\xa0-rich high fat diet in low-density lipoprotein receptor-deficient mice.

Either lipopolysaccharide (LPS) or high-fat diet (HFD) enriched with saturated fatty (SFA) promotes atherosclerosis. In this study, we investigated the effect of LPS in combination with SFA-rich HFD on atherosclerosis and how LPS and SFA interact to stimulate inflammatory response in vascular endothelial cells.Low-density lipoprotein receptor-deficient (LDLR) mice were fed a low-fat diet (LFD), HFD with low (PA) (LP-HFD), or HFD with high PA (HP-HFD) for 20 weeks. During the last 12 weeks, half mice received LPS and half received PBS. After treatment, metabolic parameters and aortic atherosclerosis were analyzed. To understand the underlying mechanisms, human aortic endothelial cells (HAECs) were treated with LPS and/or PA and proinflammatory molecule expression was quantified.The metabolic study showed that LPS had no significant effect on cholesterol, triglycerides, free fatty acids, but increased insulin and insulin resistance. Both LP-HFD and HP-HFD increased and cholesterol while LP-HFD increased glucose and HP-HFD increased triglycerides, insulin, and insulin resistance. Analysis of aortic atherosclerosis showed that HP-HFD was more effective than LP-HFD in inducing atherosclerosis and LPS in combination with HP-HFD increased atherosclerosis in the thoracic aorta, a less common site for atherosclerosis, as compared with LPS or HP-HFD. To understand the mechanisms, results showed that LPS and PA synergistically upregulated adhesion molecules and proinflammatory cytokines in HAECs.LPS and PA-rich HFD cooperatively increased atherogenesis in the thoracic aorta. The synergy between LPS and PA on proinflammatory molecules in HAECs may play an important role in atherogenesis.Published by Elsevier B.V.

Keyword: weight

Nutritional assessment, phytochemical composition and antioxidant analysis of the pulp and seed of medjool date grown in Mexico.

The aim of this study was the characterization of fatty acids, antioxidant activity, some physical properties, nutrient content, sugars, and minerals in the pulp and seeds of the date cultivar \'Medjool\' ( L.) grown in Mexico. The samples were obtained at maturity (Tamar) in the 2017 harvest season in the valleys of San Luis Rio Colorado and Mexicali, Mexico. The following average values were obtained on a % dry basis for pulp and seeds, respectively: protein, 3.14% and 4.84%; lipids, 0.75% and 9.94%; fiber, 6.34% and 66.79%; total sugars, 75.32% and 5.88%; reducing sugars, 70.26% and 4.40%; and sucrose, 5.06% and 1.46%. Analysis of the minerals revealed that the most abundant elements for the pulp were: potassium, 851.98\xa0mg/100\xa0g; magnesium, 142.97 mg/100 g; and phosphorus, 139.40 mg/100 g, whereas for the seeds, they were potassium, 413.36 mg/100 g; sulfur, 151.36 mg/100 g; and phosphorus, 92.42 mg/100 g. Gas chromatography-mass spectrometry analysis revealed that the major unsaturated fatty was oleic , at 52.34% and 45.92%, respectively, for pulp and seeds. The main saturated fatty acids were (6.75%) and lauric (17.24%) in pulp and seeds, respectively. The total phenolic content was 1.16 and 13.73 mg GAE/100 g for pulp and seeds, respectively. Finally, the antioxidant activities were: b-carotene, 65.50% and 47.75%; DPPH, 0.079 IC g/L and 0.0046 IC g/L; and ABTS, 13.72 IC g/L and 0.238 IC g/L, respectively. The results obtained in this study confirm that the \'Medjool\' cultivar grown in Mexico has the same quality of nutrients and antioxidants as those grown in the other main date-producing countries.

Keyword: weight

Perivascular adipose tissue dysfunction aggravates adventitial remodeling in obese mini pigs via NLRP3 inflammasome/IL-1 signaling pathway.

Perivascular adipose tissue (PVAT), a special type of adipose tissue, closely surrounds vascular adventitia and produces numerous bioactive substances to maintain vascular homeostasis. PVAT dysfunction has a crucial role in regulating vascular remodeling, but the exact mechanisms remain unclear. In this study, we investigated whether and how obesity-induced PVAT dysfunction affected adventitia remodeling in early vascular injury stages. Mini pigs were fed a high sugar and fat diet for 6 months to induce metabolic syndrome and obesity. In the mini pigs, left carotid vascular injury was then generated using balloon dilation. Compared with normal mini pigs, obese mini pigs displayed significantly enhanced vascular injury-induced adventitial responses, evidenced by adventitia fibroblast (AF) proliferation and differentiation, and adventitia fibrosis, as well as exacerbated PVAT dysfunction characterized by increased accumulation of resident macrophages, particularly the M1 pro-inflammatory phenotype, increased expression of leptin and decreased expression of adiponectin, and production of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Primary AFs cultured in PVAT-conditioned medium from obese mini pigs also showed significantly increased proliferation and differentiation. We further revealed that activated nod-like receptor protein 3 (NLRP3) inflammasome and its downstream products, i.e., IL-1 family members such as IL-1β and IL-18 were upregulated in the PVAT of obese mini pigs; PVAT dysfunction was also demonstrated in preadipocytes treated with . Finally, we showed that pretreatment with IL-1 receptor (IL-1R) antagonist or IL-1R knockdown blocked AF proliferation and differentiation in AFs cultured in PVAT-conditioned medium. These results demonstrate that obesity-induced PVAT dysfunction aggravates adventitial remodeling after early vascular injury with elevated AF proliferation and differentiation via activating the NLRP3/IL-1 signaling pathway.

Keyword: weight

Inhibition of hepatocyte nuclear factor 1b induces hepatic steatosis through DPP4/NOX1-mediated regulation of superoxide.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disorder that is closely associated with insulin resistance and type 2 diabetes. Previous studies have suggested that hepatocyte nuclear factor 1b (HNF1b) ameliorates insulin resistance. However, the role of HNF1b in the regulation of lipid metabolism and hepatic steatosis remains poorly understood. We found that HNF1b expression was decreased in steatotic livers. We injected mice with lentivirus (LV) expressing HNF1b shRNA to generate mice with hepatic knockdown of HNF1b. We also injected high fat (HF) diet-induced obese and db/db diabetic mice with LV expressing HNF1b to overexpress HNF1b. Knockdown of HNF1b increased hepatic lipid contents and induced insulin resistance in mice and in hepatocytes. Knockdown of HNF1b worsened HF diet-induced increases in hepatic lipid contents, liver injury and insulin resistance in mice and PA-induced lipid accumulation and impaired insulin signaling in hepatocytes. Moreover, overexpression of HNF1b alleviated HF diet-induced increases in hepatic lipid content and insulin resistance in mice. Knockdown of HNF1b increased expression of genes associated with lipogenensis and endoplasmic reticulum (ER) stress. DPP4 and NOX1 expression was increased by knockdown of HNF1b and HNF1b directly bound with the promoters of DPP4 and NOX1. Overexpression of DPP4 or NOX1 was associated with an increase in lipid droplets in hepatocytes and decreased expression of DPP4 or NOX1 suppressed the effects of knockdown of HNF1b knockdown on triglyceride (TG) formation and insulin signaling. Knockdown of HNF1b increased superoxide level and decreased glutathione content, which was inhibited by downregulation of DPP4 and NOX1. N-acetylcysteine (NAC) suppressed HNF1b knockdown-induced ER stress, TG formation and insulin resistance. (PA) decreased HNF1b expression which was inhibited by NAC. Taken together, these studies demonstrate that HNF1b plays an essential role in controlling hepatic TG homeostasis and insulin sensitivity by regulating DPP4/NOX1mediated generation of superoxide.Copyright © 2017 Elsevier Inc. All rights reserved.

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Proteomic effects of repeated-dose oral exposure to 2-monochloropropanediol and its dipalmitate in rat testes.

2- and 3-monochloropropanediol (2-MCPD) and their fatty esters are food contaminants which are concomitantly formed upon thermal treatment of foodstuff containing fats and salt. Exposure to 2- or 3-MCPD thus results, for example, from refined vegetable oils, in instant meals or infant formula, as well as in cereals or pastries. The molecular mechanisms of 2-MCPD toxicity are poorly understood. Here, we performed a comprehensive proteomic analysis of 2-MCDP-induced alterations in the testes from rats following oral administration of 10\u202fmg/kg per day 2-MCPD, or an equimolar dose of 2-MCPD dipalmitate as a representative 2-MCPD fatty ester. In the absence of overt histopathologically detectable toxicity, moderate alterations in cellular proteomic signatures were recorded. The observations are in line with the assumption that the molecular mechanisms of 2-MCPD and 3-MCPD toxicity differ. Observed proteomic alterations point towards effects of 2-MCPD on mitogen-dependent signaling and mitochondrial energy utilization. Presented data for the first time provide insight into proteomic effects of 2-MCPD in testicular tissue.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: weight

Toxicological evaluation of 2-dodecylcyclobutanone, a unique radiolytic compound of .

This study was conducted to evaluate the toxic effects and potency of 2-dodecylcyclobutanone (2-dDCB), a unique compound derived from via irradiation. In a series of assays of bacterial reverse-mutation, in vitro chromosomal aberration, and in vivo micronucleus, negative responses were found by the treatment of 2-dDCB comparing vehicle control, dimethyl sulfoxide or corn oil. In the acute oral toxicity test, all of the mice administrated 2-dDCB survived, and there were no clinical and necropsy signs observed at any doses (0, 300, and 2000\u202fmg/kg ) during the experimental period of 14 days. These results suggested that 2-dDCB is a relatively non-toxic substance with median lethality dose higher than 2000\u202fmg/kg . Moreover, there were no adverse effects noted in rats orally administrated 2-dDCB everyday via gavage for 28 days, even at the highest dose (2.0\u202fmg/kg /day) tested, which is 1000-times higher than the human daily intake of 2-dDCB estimated through an extreme exposure scenario. Overall, these results indicate that 2-dDCB is not likely to raise any human health concerns and irradiated foods containing can be recognized as safe for human consumption under the current international regulation systems for food irradiation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: weight

LC-MS/MS analysis of free fatty composition and other lipids in skins and seeds of Vitis vinifera grape cultivars.

Lipids are important constituents of plant and animal cells, being essential due to their biological functions. Despite the significant role of these compounds in grape berries, knowledge of grape lipid composition is still limited. This study addresses the free fatty composition and other lipids in skins and seeds of several Vitis vinifera grape cultivars using LC-MS/MS. A different profile of free fatty acids was determined in the grape tissues, showing a higher proportion of unsaturated fatty acids in seeds (ca. 60%). Lignoceric was one of the main free saturated fatty acids found in grape skins, together with and stearic acids. Berry skins showed similar ranges of C18-unsaturated fatty acids, whereas linoleic was predominant in the seed fatty composition. Higher content of glycerophospholipids was determined in grape seeds. Uvaol and oleanolic were only quantified in skins (1.5-3.9 and 38.6-57.6\u202fmg/kg fresh , respectively). These preliminary results suggest a certain diversity in grape lipids according to their location in the berry tissues and cultivar, providing useful information for the fermentation process due to their role in wine sensory profile and yeast growth.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: weight

Cardiomyocyte-specific knockout of endothelin receptor a attenuates obesity cardiomyopathy.

Endothelin (ET)-1 is implicated in the pathophysiology of cardiovascular diseases although its role in obesity anomalies has not been fully elucidated. This study was designed to examine the impact of ET-1 receptor A (ET) ablation on obesity-induced changes in cardiac geometry and contractile function, as well as the mechanisms involved with a focus on autophagy. Cardiomyocyte-specific ET receptor knockout (ETAKO) and WT mice were fed either low-fat (10% calorie from fat) or high-fat (45% calorie from fat) diet for 24\u202fweeks. Glucose tolerance test was examined to confirm insulin resistance. High-fat diet intake compromised myocardial geometry (enlarged left ventricular diameters in systole and diastole), morphology (cardiac hypertrophy, increased wall thickness and interstitial fibrosis), contractile function (reduced fractional shortening, ejection fraction and cardiomyocyte shortening) and intracellular Ca handling, the effect of which was significantly attenuated by ETAKO. TUNEL staining revealed overt apoptosis in high-fat-fed group, the effect was reverted by ETAKO. Western blot analysis noted that high-fat intake downregulated leptin receptor and PPARγ, insulin signaling (elevated basal/dampened insulin-stimulated phosphorylation of Akt and IRS1), phosphorylation of AMPK, ACC, upregulated GATA-4, ANP, NFATc3, PPARα, m-TOR/p70s6k signaling, which were attenuated by ETAKO with the exception of AMPK/ACC. Furthermore, high-fat intake suppressed cardiac autophagy, which was abrogated by ETAKO. In cultured murine cardiomyocytes, challenged mimicked high-fat diet-induced hypertrophic and autophagic responses, the effect of which were abolished by the ET receptor antagonist BQ123 or mTOR inhibitor rapamycin. These results suggest that inhibition of ET rescues high-fat intake-induced cardiac anomalies possibly through autophagy regulation.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: weight

Individual fatty acids in erythrocyte membranes are associated with several features of the metabolic syndrome in obese children.

Obesity leads to the clustering of cardiovascular (CV) risk factors and the metabolic syndrome (MetS) also in children and is often accompanied by non-alcoholic fatty liver disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 fatty acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting insulin (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting insulin (r\u2009=\u20090.287). Fatty Liver Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

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Ginsenoside Rb1 improves leptin sensitivity in the prefrontal cortex in obese mice.

Obesity impairs leptin-induced regulation of brain-derived neurotrophic factor (BDNF) expression and synaptogenesis, which has been considered to be associated with the incidence of neuronal degenerative diseases, cognitive decline, and depression. Ginsenoside Rb1 (Rb1), a major bioactive component of ginseng, is known to have an antiobesity effect and improve cognition. This study examined whether Rb1 can improve central leptin effects on BDNF expression and synaptogenesis in the prefrontal cortex during obesity using an in vivo and an in vitro model.Ginsenoside Rb1 (Rb1) chronic treatment improved central leptin sensitivity, leptin-JAK2-STAT3 signaling, and leptin-induced regulation of BDNF expression in the prefrontal cortex of high-fat diet-induced obese mice. In cultured prefrontal cortical neurons, , the saturated fat, impaired leptin-induced BDNF expression, reduced the immunoreactivity and mRNA expression of synaptic proteins, and impaired leptin-induced neurite outgrowth and synaptogenesis. Importantly, Rb1 significantly prevented these pernicious effects induced by .These results indicate that Rb1 reverses central leptin resistance and improves leptin-BDNF-neurite outgrowth and synaptogenesis in the prefrontal cortical neurons. Thus, Rb1 supplementation may be a beneficial avenue to treat obesity-associated neurodegenerative disorders.© 2017 John Wiley & Sons Ltd.

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Fatty Acids as Useful Serological Markers for Crohn\'s Disease.

We have previously reported that patients with Crohn\'s disease (CD) have a very specific erythrocyte membrane phospholipid fatty profile. The findings of this study suggest that the activities of enzymes involved in the metabolism of linoleic (LA), that is, delta-6 desaturase, are higher in CD patients than in healthy individuals.We evaluated the utilities of various fatty compositions of the plasma (p-) as new serological markers for CD compared to those of erythrocyte membranes (e-).Fifty CD patients and 50 healthy individuals were enrolled. In both plasma and erythrocyte membranes, the percentages of (PA) were significantly higher, while those of LA were significantly lower in CD patients than in controls. Fatty acids with high sensitivity and specificity were p-PA (0.86 and 0.74) and e-PA (0.80 and 0.74). With PA and LA as a CD fatty index (CDFAi), that is, CDFAi = (PA/LA), the sensitivity and specificity of plasma CDFAi (p-CDFAi) and e-CDFAi were 0.80 and 0.80; and 0.82 and 0.88 respectively.In CD patients, various fatty acids were specifically altered in both plasma and erythrocytes, and p-PA and p-CDFAi are potentially useful as new serological markers for CD.© 2017 S. Karger AG, Basel.

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Effects of quantitative feed restriction and sex on carcass traits, meat quality and meat lipid profile of Morada Nova lambs.

An experiment was conducted to evaluate the effects of feed restriction (FR) and sex on the quantitative and qualitative carcass traits of Morada Nova lambs. Thirty-five animals with an initial of 14.5\u2009±\u20090.89\xa0kg and age of 120 d were used in a completely randomized study with a 3\u2009×\u20093 factorial scheme consisting of three sexes (11 entire males, 12 castrated males and 12 females) and three levels of feeding (ad libitum - AL and 30% and 60% FR).Entire males presented greater hot and cold carcass (\u2009<\u20090.05), followed by castrated males and females. However, the hot carcass yield was higher for females and castrated males than for entire males. Luminosity values were influenced (\u2009<\u20090.05) by sex, with entire males presenting higher values than castrated males and females. Females showed higher (\u2009<\u20090.05) concentrations of linoleic and arachidonic in the meat of the muscle. The meat of animals submitted to AL intake and 30% FR showed similar (\u2009>\u20090.05) concentrations, and the concentrations of , palmitoleic , stearic , oleic and conjugated linoleic were higher (\u2009<\u20090.05) than those of animals with 60% FR. The meat of females had a higher ω6/ω3 ratio and lower h/H ratio, and females had greater levels of feeding. The meat of animals on the 60% FR diet had a greater ω6/ω3 ratio, lower h/H ratio and lower concentration of desirable fatty acids in addition to a greater atherogenicity index (AI) and thrombogenicity index (TI).Lambs of different sexes had carcasses with different quantitative traits without total influence on the chemical and physical meat characteristics. The lipid profile of the meat was less favorable to consumer health when the animals were female or submitted to 60% feed restriction.

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Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated inflammation.

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The protein-sparing effect of α-lipoic in juvenile grass carp, Ctenopharyngodon idellus: effects on lipolysis, fatty β-oxidation and protein synthesis.

To investigate the protein-sparing effect of α-lipoic (LA), experimental fish (initial : 18·99 (sd 1·82) g) were fed on a 0, 600 or 1200 mg/kg α-LA diet for 56 d, and hepatocytes were treated with 20 μm compound C, the inhibitor of AMP kinase α (AMPKα), treated for 30 min before α-LA treatment for 24 h. LA significantly decreased lipid content of the whole and other tissues (P0·05). Consistent with results from the experiment in vitro, LA activated phosphorylation of AMPKα and notably increased the protein content of adipose TAG lipase in intraperitoneal fat, hepatopancreas and muscle in vivo (P<0·05). Meanwhile, LA significantly up-regulated the mRNA expression of genes involved in fatty β-oxidation in the same three areas, and LA also obviously down-regulated the mRNA expression of genes involved in amino catabolism in muscle (P<0·05). Besides, it was observed that LA significantly activated the mammalian target of rapamycin (mTOR) pathway in muscle of experimental fish (P<0·05). LA could promote lipolysis and fatty β-oxidation via increasing energy supply from lipid catabolism, and then, it could economise on the protein from energy production to increase protein deposition in grass carp. Besides, LA might directly promote protein synthesis through activating the mTOR pathway.

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Recycling of food waste for fuel precursors using an integrated bio-refinery approach.

The main aim was to integrate FW-recycling with cultivation of Rhodotorula glutinis and anaerobic digestion (AD) for bio-energy and -fuel recovery. Mixed FW was mechanically macerated (Pcon) and hydrolysed (at 250gL water) via chemical (Ch), thermal (Th) and TCh (combined Ch and Th) treatments. Cleared hydrolysates from individual pre-treatment processes were used as culture medium for cultivation of R. glutinis, while the residual solids (RS) were subjected to AD. Pcon cultivation yielded maximal R. glutinis dry biomass (5.18gL) and total fatty contents (1.03gg DW). Dominant fatty methyl esters (FAME) were - (C-26%); Stearic - (C-17%) and Oleic acids (C-38%), ideal for bio-diesel production. Highest methane yields (actual ∼0.139mkg volatile solids) were measured from AD of Th-derived RS. Thus integrated FW recycling approaches will be more feasible for generating energy and economic incentives.Copyright © 2017 Elsevier Ltd. All rights reserved.

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Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to- ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

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Palmitate aggravates proteinuria-induced cell death and inflammation via CD36-inflammasome axis in the proximal tubular cells of obese mice.

High levels of serum free fatty acids (FFAs) and proteinuria have been implicated in the pathogenesis of obesity-related nephropathy. CD36, a class B scavenger receptor, is highly expressed in the renal proximal tubules and mediates FFA uptake. It is not clear whether FFA- and proteinuria-mediated CD36 activation coordinates NLRP3 inflammasomes to induce renal tubular injury and inflammation. In this study, we investigated the roles of CD36 and NLRP3 inflammasomes in FFA-induced renal injury in high-fat diet (HFD)-induced obesity. HFD-fed C57BL/6 mice and palmitate-treated HK2 renal tubular cells were used as in vivo and in vitro models. Immunohistochemical staining showed that CD36, IL-1β, and IL-18 levels increased progressively in the kidneys of HFD-fed mice. Sulfo- N-succinimidyl oleate (SSO), a CD36 inhibitor, attenuated the HFD-induced upregulation of NLRP3, IL-1β, and IL-18 and suppressed the colocalization of NLRP3 and ASC in renal tubular cells. In vitro, SSO abolished the palmitate-induced activation of IL-1β, IL-18, and caspase-1 in HK2 proximal tubular cells. Furthermore, treatment with SSO and the knockdown of caspase-1 expression by siRNA both inhibited palmitate-induced cell death and apoptosis in HK2 cells. Collectively, palmitate causes renal tubular inflammation, cell death, and apoptosis via the CD36/NLRP3/caspase-1 axis, which may explain, at least in part, the mechanism underlying FFA-related renal tubular injury. The blockade of CD36-induced cellular processes is therefore a promising strategy for treating obesity-related nephropathy.

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The effect of surfactant crystallization on partial coalescence in O/W emulsions.

Partial coalescence is a ubiquitous instability in emulsions whose dispersed phase is partially crystallized. When emulsions are stabilized with proteins, interfacial stiffness and long-range repulsive surface forces hinder this type of instability. The addition of low molecular surfactants modifies the interfacial properties and surface forces, generally promoting partial coalescence. In the present work, various surfactants (Tween® 80, and monoglycerides) differing in their crystallization temperature were probed for their ability to induce partial coalescence in model O/W emulsions stabilized by sodium caseinate. The initially fluid emulsions were submitted to a tempering cycle leading to the gelation of the system. The extent of partial coalescence was evaluated by measuring the bulk storage modulus. DSC was used to determine the melting range of the oil phase and surfactants, while polarized microscopy, Raman imaging, and surface rheology measurements were performed to characterize the oil/water interface. The experimental conditions in terms of droplet size, surfactant-to-protein molar ratio and tempering history favoring partial coalescence were first explored in presence of Tween® 80. We show that partial coalescence is rather marginal when crystallizable surfactants are added, and pronounced with liquid surfactants. The phenomena underlying this result, especially interfacial crystallization of surfactants, are evidenced and discussed.Copyright © 2017 Elsevier Inc. All rights reserved.

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Ellagic Suppresses the Oxidative Stress Induced by Dietary-Oxidized Tallow.

Dietary tallow was thermally oxidized at 180°C in an open fryer. The oxidized tallow (OT) and unoxidized tallow were characterized for oxidation parameters and fatty composition using GC-MS. Tallow samples were fed to rabbits along with 50, 100, and 150\u2009mg/kg/day of ellagic (EA) for three weeks. Results revealed that the peroxide value (PV) and thiobarbituric reactive substances (TBARS) significantly increased, while radical scavenging activity (RSA) of the tallow decreased significantly with oxidation. GC-MS analysis showed eight fatty acids in the tallow samples, where (48.5-49.7\u2009g/100\u2009g), linoleic (18.7-23.7\u2009g/100\u2009g), stearic (13.5-15.6\u2009g/100\u2009g), and margaric (6.32-6.42\u2009g/100\u2009g) were the major fatty acids. Animal studies showed that oxidized tallow (OT) alone or in combination with EA significantly altered the of the rabbits. Serum biochemical parameters and renal function tests were affected by OT and ameliorated by EA. The toxic effects of OT on haematological indices were minimized by EA. The supplementation of OT alone had significant effects on the liver structure and functions. The coadministration of EA reduced the toxic properties of OT on the liver, by increasing the antioxidant (GSH) system. The rabbit heart was also affected by the OT, which was ameliorated by EA supplementation. These results suggested that the supplementation of EA was beneficial against the OT-induced oxidative stress and may be considered for foods containing oxidized lipids.

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Postprandial Lipid Response to High-Saturated and High-Monounsaturated Fat Meals in Normal- or Overweight Women.

We evaluated postprandial response of the lipid metabolism markers after the intake of a high-saturated fat (HSM) or high-monounsaturated fat meal (HMM).A randomized, controlled and acute intervention study included 63 women (age 26.9 ± 6.1 years): 35 normal (NW) and 28 overweight (OW) (total fat [TBF] 24.7 ± 3.9% and 36.6 ± 3.9%, respectively). After 12\xa0hours of fasting, each subject was given one of the two test meals standardized, including 2 muffins and water (HSM, 42.1% of saturated fat , or HMM, 34.5% of monounsaturated fat ). Plasma fatty profile and concentrations of apolipoproteins A1 and B100, complement C3, and triacylglycerols were analyzed during fasting and at 2, 3, and 5 postprandial hours.Among the markers studied, the triacylglycerol (TAG) and complement C3 were significantly higher in the OW group, compared to NW. The increment in the C3 concentration was higher after HSM intake, compared with HMM (iAUC = 4365.5 ± 5477.4\xa0vs. 1215.2 ± 882.4; p = 0.006), with no differences between groups. After 5\xa0hours postprandial, plasma oleic values remained high compared with the fasting value in the NW group, but not in the OW group (26.0 ± 4.2\xa0vs 23.7 ± 3.9%; p < 0.001). Women with high percentage of total plasma saturated fatty acids (SFA) at the beginning of the intervention had higher incremental area under the curve (iAUC) for the , stearic, and total fatty acids (p < 0.005). Those women with a high percentage of monounsaturated fatty acids (MUFA) showed lower iAUC values for the same fatty profile (p < 0.005).This study demonstrated the effect of the HSM on postprandial increment of C3 concentration, suggesting another mechanism for saturated fat metabolism. The postprandial response to HSM appears to be the mediated by baseline lipid profile of the individuals, while the response to HMM was correlated to the status.

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Inhibition of central de novo ceramide synthesis restores insulin signaling in hypothalamus and enhances β-cell function of obese Zucker rats.

Hypothalamic lipotoxicity has been shown to induce central insulin resistance and dysregulation of glucose homeostasis; nevertheless, elucidation of the regulatory mechanisms remains incomplete. Here, we aimed to determine the role of de novo ceramide synthesis in hypothalamus on the onset of central insulin resistance and the dysregulation of glucose homeostasis induced by obesity.Hypothalamic GT1-7 neuronal cells were treated with palmitate. De novo ceramide synthesis was inhibited either by pharmacological (myriocin) or molecular (si-Serine Palmitoyl Transferase 2, siSPT2) approaches. Obese Zucker rats (OZR) were intracerebroventricularly infused with myriocin to inhibit de novo ceramide synthesis. Insulin resistance was determined by quantification of Akt phosphorylation. Ceramide levels were quantified either by a radioactive kinase assay or by mass spectrometry analysis. Glucose homeostasis were evaluated in myriocin-treated OZR. Basal and glucose-stimulated parasympathetic tonus was recorded in OZR. Insulin secretion from islets and β-cell mass was also determined.We show that palmitate impaired insulin signaling and increased ceramide levels in hypothalamic neuronal GT1-7 cells. In addition, the use of deuterated demonstrated that palmitate activated several enzymes of the de novo ceramide synthesis pathway in hypothalamic cells. Importantly, myriocin and siSPT2 treatment restored insulin signaling in palmitate-treated GT1-7 cells. Protein kinase C (PKC) inhibitor or a dominant-negative PKCζ also counteracted palmitate-induced insulin resistance. Interestingly, attenuating the increase in levels of hypothalamic ceramides with intracerebroventricular infusion of myriocin in OZR improved their hypothalamic insulin-sensitivity. Importantly, central myriocin treatment partially restored glucose tolerance in OZR. This latter effect is related to the restoration of glucose-stimulated insulin secretion and an increase in β-cell mass of OZR. Electrophysiological recordings also showed an improvement of glucose-stimulated parasympathetic nerve activity in OZR centrally treated with myriocin.Our results highlight a key role of hypothalamic de novo ceramide synthesis in central insulin resistance installation and glucose homeostasis dysregulation associated with obesity.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

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Synthesis of chemically edited derivatives of the endogenous regulator of inflammation 9-PAHSA.

Fatty esters of hydroxy fatty acids (FAHFAs) are a growing class of natural products found in organisms ranging from plants to humans. The roles these endogenous derivatives of fatty acids play in biology and their novel pathways for controlling inflammation have increased our understanding of basic human physiology. FAHFAs incorporate diverse fatty acids into their structures, however, given their recent discovery non-natural derivatives have not been a focus and as a result structure-activity relationships remain unknown. The importance of the long chain hydrocarbons extending from the ester linkage as they relate to anti-inflammatory activity is unknown. Herein the systematic removal of carbons from either the hydroxy fatty or fatty regions of the most studied FAHFA, ester of 9-hydroxystearic (9-PAHSA), was achieved and these synthetic, abridged analogs were tested for their ability to attenuate IL-6 production. Reduction of the carbon chain lengths of the 9-hydroxystearic portion or hydrocarbon chain resulted in lower molecular analogs that maintained anti-inflammatory activity or in one case enhanced activity.

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Chitosan Oligosaccharides Improve Glucolipid Metabolism Disorder in Liver by Suppression of Obesity-Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid metabolism disorder using (PA)-induced HepG2 cells and high-fat diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated lipid accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid metabolism-related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid metabolism disorder in mice fed with HFD for five months, including reduction in and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid metabolism-related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid metabolism disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid metabolism-related diseases.

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GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

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Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with metabolic syndrome.

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an energy-restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with metabolic syndrome.The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of metabolic syndrome.After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk energy-restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

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GLP-1 Analogue, Exendin-4, Modulates MAPKs Activity but not the Heat Shock Response in Human HepG2 Cells.

Glucagon-like peptide-1 (GLP-1) analogues reduce ER stress and inflammation in key metabolic organs, including the liver. However, their effects on heat shock response (HSR) and mitogen-activated protein kinases (MAPKs) have not yet been elucidated. In the present study, we investigate whether the GLP-1 analogue, exendin-4, triggers the expression of HSR and increases MAPK activity under metabolic stress.The effects of exendin-4 in the presence or absence of (PA; 400 μm) or glucose (30 mm) in the HepG2 liver cell line are assessed using Western blots, quantitative real-time PCR, and label-free proteomics.Heat shock proteins (HSP60, HSP72, HSP90, and GRP78) and other chaperones are not significantly affected by exendin-4 under the conditions tested. In contrast, the presence of exendin-4 alone increases the MAPK phosphorylation levels (JNK, ERK1/2, and p38). For short incubation periods, in the presence of PA or glucose, treatment with exendin-4 exhibits limited effects but significantly attenuates MAPK phosphorylation after a 24-h incubation. Interestingly, canonical signaling pathways, such as EIF2, ILK, PKA, and Rho, are modulated by exendin-4.Identifying new pathways modulated by GLP-1 analogues will provide further insights into their benefits beyond their currently recognized roles in glycemic control, such as MAPK activity, energy homeostasis, and decrease.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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Rational design of dimeric lipidated Xenopus glucagon-like peptide 1 analogues as long-acting antihyperglycaemic agents.

Dimerization is viewed as an effective means to enhance the binding affinity and therapeutic potency of peptides. Both dimerization and lipidation effectively prolong the half-life of peptides in\xa0vivo by increasing hydrodynamic size and facilitating physical interactions with serum albumin. Here, we report a novel method to discover long-acting glucagon-like peptide 1 (GLP-1) analogues by rational design based on Xenopus GLP-1 through a combined dimerization and lipidization strategy. On the basis of our previous structure analysis of Xenopus GLP-1, and a C-terminal Cys were firstly introduced into two Xenopus GLP-1 analogues (1 and 2), and the afforded 3 and 4 were further reacted with bis-maleimide amine to afford two dimeric lipidated Xenopus GLP-1 analogues (5 and 6). The in\xa0vitro and in\xa0vivo biological activities of 5 and 6 were significantly improved as compared with their monomers. Moreover, the selected compound 6 showed greater hypoglycemic and insulinotropic activities than liraglutide even when the dose of 6 was reduced to half in db/db mice. Pharmacokinetic test revealed that 6 had a ∼ 3-fold longer half-life than liraglutide in Kunming mice and SD rats, and the longer half-life of 6 led to excellent long-acting hypoglycemic effects as confirmed by two different pharmacological methods conducted on db/db mice. Finally, a 7 weeks chronic study conducted on db/db mice demonstrated the better therapeutic efficacies of 6 on glucose tolerance normalization, HbA1c reduction and pancreas islets protection than liraglutide. The present research showed that combined dimerization and lipidization is effective when applied to Xenopus GLP-1 analogue to develop novel GLP-1 analogue for the treatment of type 2 diabetes. In addition, the promising preclinical data of 6 suggested the therapeutic potential of 6 as a novel anti-diabetic agent.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

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Vildagliptin Can Alleviate Endoplasmic Reticulum Stress in the Liver Induced by a High Fat Diet.

We investigated whether a DDP-4 inhibitor, vildagliptin, alleviated ER stress induced by a high fat diet and improved hepatic lipid deposition. C57BL/6 mice received standard chow diet (CD), high fat diet (HFD), and HFD administered with vildagliptin (50\u2009mg/Kg) (V-HFD). After administration for 12 weeks, serum alanine aminotransferase, glucose, cholesterol, triglyceride, and insulin levels were analyzed. Samples of liver underwent histological examination and transmission electron microscopy, real-time PCR for gene expression levels, and western blots for protein expression levels. ER stress was induced in HepG2 cells with and the effects of vildagliptin were investigated. HFD mice showed increased liver / (20.27%) and liver triglycerides (314.75%) compared to CD mice, but these decreased by 9.27% and 21.83%, respectively, in V-HFD mice. In the liver, HFD induced the expression of ER stress indicators significantly, which were obviously decreased by vildagliptin. In vitro, the expressions of molecular indicators of ER stress were reduced in HepG2 when vildagliptin was administered. Vildagliptin alleviates hepatic ER stress in a mouse high fat diet model. In HepG2 cells, vildagliptin directly reduced ER stress. Therefore, vildagliptin may be a potential agent for nonalcoholic fatty liver disease.

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Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver- ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

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Maternal overnutrition by hypercaloric diets programs hypothalamic mitochondrial fusion and metabolic dysfunction in rat male offspring.

Maternal overnutrition including pre-pregnancy, pregnancy and lactation promotes a lipotoxic insult leading to metabolic dysfunction in offspring. Diet-induced obesity models (DIO) show that changes in hypothalamic mitochondria fusion and fission dynamics modulate metabolic dysfunction. Using three selective diet formula including a High fat diet (HFD), Cafeteria (CAF) and High Sugar Diet (HSD), we hypothesized that maternal diets exposure program leads to selective changes in hypothalamic mitochondria fusion and fission dynamics in male offspring leading to metabolic dysfunction which is exacerbated by a second exposure after weaning.We exposed female Wistar rats to nutritional programming including Chow, HFD, CAF, or HSD for 9\xa0weeks (pre-mating, mating, pregnancy and lactation) or to the same diets to offspring after weaning. We determined , food intake and metabolic parameters in the offspring from 21 to 60\xa0days old. Hypothalamus was dissected at 60\xa0days old to determine mitochondria-ER interaction markers by mRNA expression and western blot and morphology by transmission electron microscopy (TEM). Mitochondrial-ER function was analyzed by confocal microscopy using hypothalamic cell line mHypoA-CLU192.Maternal programming by HFD and CAF leads to failure in glucose, leptin and insulin sensitivity and fat accumulation. Additionally, HFD and CAF programming promote mitochondrial fusion by increasing the expression of MFN2 and decreasing DRP1, respectively. Further, TEM analysis confirms that CAF exposure after programing leads to an increase in mitochondria fusion and enhanced mitochondrial-ER interaction, which partially correlates with metabolic dysfunction and fat accumulation in the HFD and CAF groups. Finally, we identified that lipotoxic stimulus in hypothalamic cells increases Ca overload into mitochondria matrix leading to mitochondrial dysfunction.We concluded that maternal programming by HFD induces hypothalamic mitochondria fusion, metabolic dysfunction and fat accumulation in male offspring, which is exacerbated by HFD or CAF exposure after weaning, potentially due to mitochondria calcium overflux.

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Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism.A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.© 2018 The Author(s). Published by S. Karger AG, Basel.

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Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant gain differences, with the top one third gaining 48% heavier than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

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Ameliorative effect of panaxynol on the reduction in high-molecular- adiponectin secretion from 3T3-L1 adipocytes treated with acids.

Reduced plasma levels of the high-molecular (HMW) form of adiponectin, rather than total adiponectin levels, have been shown to be closely associated with various metabolic diseases including insulin resistance, type 2 diabetes, and cardiovascular disease. Therefore, we sought to explore active, naturally occurring compounds that promote the recovery of HMW adiponectin secretion suppressed by in our model. A total of 90 crude drug extracts were screened for the ability to augment HMW adiponectin secretion from 3T3-L1 adipocytes treated with . Panaxynol was isolated from Saposhnikovia divaricata as an active compound with HMW adiponectin promoting properties. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists are reported to increase the secretion of HMW adiponectin, although the effects of panaxynol were found to be independent of PPARγ activation. When the underlying mechanisms were further examined, panaxynol was found to inhibit the -induced downregulation of forkhead box O1 (FoxO1) protein, and the anti-lipotoxic effects were abolished by a FoxO1 inhibitor. Furthermore, CCAAT/enhancer-binding protein-α (C/EBPα) mRNA levels were also increased by panaxynol. Reactive oxygen species have critical roles in the reduction in HMW adiponection secretion by ; however, panaxynol reduced this increase in reactive oxygen species generation, followed by reductions in markers of endoplasmic reticulum stress and inflammation. Taken together, these findings suggest that panaxynol ameliorates the impaired HMW adiponection secretion in adipocytes treated with by restoring FoxO1 expression, owing to inhibition of reactive oxygen species generation, in a PPARγ-independent manner.Copyright © 2017 Elsevier B.V. All rights reserved.

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Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

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Valorization of crude glycerol based on biological processes for accumulation of lipophilic compounds.

Bacteria that are capable of accumulating lipids in their cells as storage compounds can also produce polyhydroxyalkanoates of high technological value, depending on the specific culture conditions. The objective of this study was to utilize crude glycerol from biodiesel (CGB) as a substrate, which is a major byproduct from biodiesel production, to produce lipophilic compounds. Bacillus megaterium INCQS 425 was cultivated and evaluated for the production of lipophilic compounds and the properties of these compounds were investigated. Cultivation of the bacteria in a medium with a C:N ratio of 0.60:1 favored the accumulation of lipids by (17.5%) comprising mainly (13.08%), palmitoleic (39.48%), and especially oleic (37.02%), which imparts good characteristics to biodiesel. Meanwhile, cultivation of the bacteria in a medium with a C:N ratio of 4:1 favored the accumulation of polyhydroxyalkanoates (PHA) (3.31gL) mainly comprising medium and long chain PHA. Low crystallinity (<30%) and excellent thermal properties make them suitable for processes that demand high temperatures, such as extrusion. The lipids produced in the present study had satisfactory oxidative stability for the production of quality biodiesel. The polyhydroxyalkanoates produced in the study are of low cost and have promising thermal properties that justify its technological potential, thereby configuring highly competitive bioproducts.Copyright © 2019. Published by Elsevier B.V.

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Dietary lipid requirement of lemon fin barb hybrid.

The present study has been conducted to determine the optimal dietary requirement lipid of lemon fin barb hybrid fingerlings. Five isonitrogenous diets (35% protein) with graded lipid levels of 0%, 4%, 6%, 8% and 12% were fed to 225 lemon fin barb hybrid fingerlings (4.3 ? 0.5 g), which were randomly and equally distributed into twelve 60 l aquaria. The diets were randomly assigned to the aquaria in triplicates, and the feeding experiment was conducted for 10 weeks. Fish were fed at 5% and the feeding ration was adjusted fortnightly. The gain, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) of fish fed 4% dietary lipid were significantly superior (P<0.05) than those fed with other diets, except for 6% lipid. Except for whole moisture content, there were significant differences (P<0.05) between whole proximate composition and nutrient retentions of fish fed with different test diets. The results showed that was the main saturated fatty (SFA), oleic was the predominant MUFA while arachidic and docosahexanoic were the major PUFAs in the muscle and liver tissue of fish. The n-3/n-6 ratio ranged from 0.66 to 2.65. From the results of this study, a dietary lipid of 4% was recommended for lemon fin barb hybrid. ?

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Prepartum fatty supplementation in sheep. III. Effect of eicosapentaenoic and docosahexaenoic during finishing on performance, hypothalamus gene expression, and muscle fatty acids composition in lambs.

The objectives of this study were to evaluate the effect of feeding an enriched diet with eicosapentaenoic (EPA) and docosahexaenoic (DHA) to finishing lambs born from ewes supplemented either with or without EPA and DHA during late gestation on productive performance, muscle fatty (FA), and hypothalamus mRNA concentration of metabolic genes and hormone receptors. Lambs born from dams fed during the last 50 d of gestation either with a control diet containing 0.39% Ca salts of fatty distillate (C) or Ca salts enriched with EPA and DHA (PFA) were used. After weaning lambs (n = 70) were blocked by (BW) and used in a 2 × 2 factorial into 2 finishing diets containing 1.5% of C or PFA. The 2 factors were the ewe diet and the finishing diet. Lambs (37.9 ± 0.4 kg) were weighed and blood sampled for glucose and NEFA measurements at days 1, 14, 28, and 42. Dry matter intake (DMI) was measured daily. At day 43, 14 females and 14 males were slaughtered, and hot carcass , wall thickness, rib eye area, and FA composition of Longissumus thoracis muscle were evaluated. Female hypothalamuses were obtained and mRNA concentration of hormone receptors, neuropeptides, and their receptors was measured. Lambs born from PFA dams were heavier (P < 0.01). There was a time × finishing diet interaction for BW (P = 0.03), and lambs fed C had a greater BW. Lambs fed C had an increase in DMI (P < 0.01). There were no significant differences in plasma glucose and NEFA concentration (P > 0.1). Lambs born from PFA dams had a greater concentration of C22:0 (P < 0.03). Lambs fed C had higher concentrations of C18:1c15 (P < 0.01), C17:0 (P < 0.09), C18:0 (P < 0.09), and n6/n3 (P < 0.01). Lambs fed PFA had greater concentration (P < 0.05) of C16:1, C22:1, C20:5, C22:5, C22:6, total n3 FA, and total EPA and DHA. There was a significant dam × finishing diet interaction (P ≤ 0.08) on mRNA concentration for MCR3, CCK-R, Cort-R, and CART. Lambs, which had the same treatment as their dams, showed lower overall mRNA concentration than those with different treatments between them and their dams. Lambs born from PFA ewes had lower concentration of MCR4 mRNA (P = 0.09) than C. Agouti-related peptides mRNA concentration was lower in lambs fed PFA (P = 0.06) than C. In conclusion, changes on lamb performance, muscle fatty composition, and metabolic neuropeptides depend not only on the lamb diet, but also on the dam diet during late gestation.

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Nutritionally Derived Metabolic Cues Typical of the Obese Microenvironment Increase Cholesterol Efflux Capacity of Adipose Tissue Macrophages.

Cholesterol retention within plasma membranes of macrophages is associated with increased inflammatory signaling. Cholesterol efflux via the transporters ABCA1, ABCG1, and SR-BI to high-density lipoprotein (HDL) particles is a critical mechanism to maintain cellular cholesterol homeostasis. Little is known about the impact of the obese microenvironment on cholesterol efflux capacity (CEC) of macrophages. In this study, the CEC of obese-derived primary adipose-tissue macrophages (ATM) is evaluated and the in vivo microenvironment is modeled in vitro to determine mechanisms underlying modulated CEC.F4/80 ATM are labeled with H-cholesterol ex vivo, and CEC and ABCA1/ABCG1 protein levels are determined. Total, ABCA1-dependent, and ABCA1-independent CECs are determined in J774 macrophages polarized to M1 (LPS&IFNγ), M2 (IL-4&IL-13), or metabolic phenotypes (glucose, insulin, and ).Obese ATM exhibit enhanced CEC and ABCA1 and ABCG1 expression compared to lean ATM. In contrast, ABCA1-CEC is suppressed from M1 polarized macrophages compared to untreated in vitro, by activation of the JAK/STAT pathway. Incubation of macrophages in vitro in high glucose augments cAMP-induced ABCA1 protein expression and ABCA1-CEC.These novel findings demonstrate remarkable plasticity of macrophages to respond to their environment with specific modulation of ABCA1 depending on whether classical pro-inflammatory or metabolic cues predominate.© 2018 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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Nutritional and Nutraceutical Composition of Pansies (Viola × wittrockiana) During Flowering.

Edible flowers consumption and use are an increasing food trend worldwide, although information concerning their nutritional composition and nutraceutical value is still scarce. Thus, the aim of this study was to contribute to the popularization of pansies (Viola × wittrockiana), through the analysis of the nutritional and nutraceutical features of pansies with different colors (white, yellow, and red) and flowering stages. Both flower type and flowering stage influenced the flower composition. When completely open, white and yellow pansies had the highest contents of protein (>2.00\xa0g/100\xa0g fresh ), while red pansies had the highest content of carbohydrates (8.0 g/100\xa0g fresh ). Regarding the fatty profiles, linoleic was always predominant (ranging between 18.7 and 51.0 g/100\xa0g fatty acids), followed by the and linolenic acids. During flowering, there was an increase in protein, fat, and linolenic contents in white and yellow pansies, whereas in red pansies the values did not change. Red pansies were characterized by the highest contents of total carotenoids (873 to 1300 µg β-carotene/g dry ) and monomeric anthocyanins (303 to 402 µg Cy-3 glu/g dry ); however, white and yellow pansies showed an increase in the values of total reducing capacity (total phenols), hydrolysable tannins, flavonoids, monomeric anthocyanins, and antioxidant activity from the bud to completely open flower stage. Our results underline the nutritional differences between pansies with different colors at distinct stages of development and their potential health benefits, suggesting that they can be used as ingredient to improve the nutritional properties of foods. PRACTICAL APPLICATION: The market of edible flowers is increasing, although little information in nutritional view is available. So, the present study was conducted to contribute to the popularization of edible flowers as a new and prospective source for the food industry, as well as a promising product for human nutrition. The results of the present study underline the nutritional differences between pansies with different colors at distinct stages of development and their potential health benefits, suggesting that they can be used as ingredient to improve the nutritional properties of foods.© 2019 Institute of Food Technologists®.

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Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid metabolism-related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of lipid metabolism- and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid metabolism-related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid metabolism, suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

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Supplemental flaxseed modulates ovarian functions of weanling gilts via the action of selected fatty acids.

The aim of this study was to examine the influence of dietary flaxseed on the endocrine and ovarian functions of weanling gilts challenged with E. coli and Coronavirus infections treated with dietary probiotic cheeses and to understand the possible mechanisms of its effects on ovarian function. Probiotics were used as a natural substitution for antibiotics and 10% dietary flaxseed is an effective prebiotic which supports the action of probiotics and has other beneficial effects on the organism. Probiotics with or without flaxseed were fed to weanling gilts starting 10 days before and lasting up until 14 days after weaning. The ovaries were measured and histologically analysed. The blood samples for the levels of steroid hormones and insulin-like growth factor I (IGF-I) were assessed using immunoassays and the levels of fatty acids were assessed using gas chromatography. All samples were collected on the day of weaning and 14 days after weaning. On the day of weaning, increased levels of linoleic and IGF-I was associated with higher . The steroid hormones were not affected by the diet. The conversion of alpha-linolenic (ALA) to timodonic (EPA) and cervonic (DHA) acids were lower compared to controls, and together with high levels of myristic, and palmitoleic acids was associated with the higher proliferation and lower apoptosis in the primordial, primary and secondary follicles; although the inhibition of the cell cycle was observed in relation to the low level of eicosadienoic . The high levels of ALA, EPA and DHA and the low levels of myristic, and palmitoleic acids may have been the effect of flaxseed feeding 14 days post-weaning and may have had a reverse effect on the proliferation and apoptosis of ovarian follicles. These data suggest that flaxseed may suppress the follicle development in weanlings via the stimulation of apoptosis and the inhibition of proliferation via the modulation of the metabolism of selected fatty acids.Copyright © 2018 Elsevier B.V. All rights reserved.

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Performances of local poultry breed fed black soldier fly larvae reared on horse manure.

In poultry, feed based on maggots, like larvae of black soldier fly () is an attractive option to substitute current ingredients which are expensive and often in direct or indirect competition with human food. Little information is currently available on the utility of these larvae in poultry feed, so goals of this study were to determine whether larvae could be reared on horse manure under traditional farming conditions and to evaluate the growth performances of a local poultry fed these larvae and the fatty acids profiles of their meat. After freezing and thawing, larvae were introduced in the feed of Ardennaise chickens between 30 and 80 days of age. Birds in the control group received a commercial standard feed, while those in the treatment group received the same commercial feed in which 8% was substituted with whole fresh larvae corresponding to 2% on a dry matter basis. Means ± standard errors of larval length and were 20.67\xa0±\xa02.21\xa0mm and 0.14\xa0±\xa00.02\xa0g, respectively. Mean larval percentages of dry matter and of substances extractable in diethyl ether were 24.6% and 23.1%, respectively. Larval fatty acids profiles were predominantly composed of lauric (28.1%) and (22.0%). Least squares means of weekly of chicken, adjusted for the effects of sex, replication and initial , were significantly higher (\xa0<\xa00.05) by 77.03\xa0±\xa053.37\xa0g in larvae-fed than in control chickens. All other measurements were not statistically different between larvae-fed and control chicken, including fatty profiles, protein content and ω6/ω3 ratio.

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Hypothalamic lipid-laden astrocytes induce microglia migration and activation.

Obesity-induced hypothalamic inflammation is closely associated with various metabolic complications and neurodegenerative disorders. Astrocytes, the most abundant glial cells in the central nervous system, play a crucial role in pathological hypothalamic inflammatory processes. Here, we demonstrate that hypothalamic astrocytes accumulate lipid droplets under saturated fatty -rich conditions, such as obese environment, and that the lipid-laden astrocytes increase astrogliosis markers and inflammatory cytokines (TNFα, IL-1β, IL-6, MCP-1) at the transcript and/or protein level. Medium conditioned by the lipid-laden astrocytes stimulate microglial chemotactic activity and upregulate transcripts of the microglia activation marker Iba-1 and inflammatory cytokines. These findings indicate that the lipid-laden astrocytes formed in free fatty -rich obese condition may participate in obesity-induced hypothalamic inflammation through promoting microglia migration and activation.© 2017 Federation of European Biochemical Societies.

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NDP-MSH reduces oxidative damage induced by in primary astrocytes.

Recent findings relate obesity to inflammation in key hypothalamic areas for control. Hypothalamic inflammation has also been related to oxidative stress. (PA) is the most abundant free fatty found in food, and in vitro studies indicate that it triggers a pro-inflammatory response in the brain. Melanocortins are neuropeptides with proven anti-inflammatory and neuroprotective action mediated by melanocortin receptor 4 (MC4R), but little is known about the effect of melanocortins on oxidative stress. The aim of this study was to investigate whether melanocortins could alleviate oxidative stress induced by a high fat diet (HFD) model. We found that NDP-MSH treatment decreased PA-induced reactive oxygen species production in astrocytes, an effect blocked by the MC4R inhibitor JKC363. NDP-MSH abolished nuclear translocation of Nrf2 induced by PA and blocked the inhibitory effect of PA on superoxide dismutase (SOD) activity and glutathione levels while it also per se increased activity of SOD and γ-glutamate cysteine ligase (γ-GCL) antioxidant enzymes. However, HFD reduced hypothalamic MC4R and brain derived neurotrophic factor mRNA levels, thereby preventing the neuroprotective mechanism induced by melanocortins.© 2019 British Society for Neuroendocrinology.

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Lipid classes and fatty acids composition of the roe of wild Silurus glanis from subalpine freshwater.

The fat content and the fatty composition of the roe of European catfish (Silurus glanis) wild specimens captured in the Lambro river basin (Como, Northern Italy) were investigated in order to assess their nutritional value. Total extracted lipid values on roe were 5.8-6.3%. Phospholipids (40.2-43.6%) and triacylglycerols (31.8-34.7%) were the most represented lipid classes. Relevant amount of cholesteryl esters (14.3-15.6%) were also detected. The main fatty acids were , oleic and cis-4,7,10,13,16,19-docosahexaenoic (DHA). Total saturated fatty acids were 28.1-30.9%, monounsaturated fatty acids were 28.2-30.6%, and polyunsaturated fatty acids (PUFAs) were 26.7-29.1%. The good found content of phospholipids and ω3-polyunsaturated fatty acids encourage further investigation for the exploitation of Silurus glanis roe in food products and/or supplements.Copyright © 2017 Elsevier Ltd. All rights reserved.

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MD2 Blockage Protects Obesity-Induced Vascular Remodeling via Activating AMPK/Nrf2.

Obesity and increased free fatty (FFA) levels are tightly linked with vascular oxidative stress and remodeling. Myeloid differentiation 2 (MD2), an important protein in innate immunity, is requisite for endotoxin lipopolysaccharide responsiveness. This study shows that (PA) also bonds to MD2, initiating cardiac inflammatory injury. However, it is not clear whether MD2 plays a role in noninflammatory systems such as obesity- and FFA-related oxidative stress involved in vascular remodeling and injury. The aim of this study is to examine whether MD2 participates in reactive oxygen species increase and vascular remodeling.Male MD2 mice and wild-type littermates with a C57BL/6 background were fed a high-fat diet (HFD) to establish obesity-induced vascular remodeling. Rat aortic endothelial cells (RAECs) and vascular smooth muscle cells (VSMCs) were treated with PA to induce oxidative stress and injury.In vivo, MD2 deficiency significantly reduced HFD-induced vascular oxidative stress, fibrosis, and remodeling, accompanied with AMP-activated kinase (AMPK) activation and nuclear factor erythroid (Nrf2) upregulation. In VSMCs and RAECs, inhibition of MD2 by neutralizing monoclonal antibody to MD2 or small interfering RNA knockdown significantly activated the AMPK/Nrf2-signaling pathway and reduced PA-induced oxidative stress and cell injury.It was demonstrated that the deletion or inhibition of MD2 protects against HFD/FFA-induced vascular oxidative stress and remodeling by activating the AMPK/Nrf2-signaling pathway.© 2017 The Obesity Society.

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Inhibition of soluble epoxide hydrolase attenuates a high-fat diet-mediated renal injury by activating PAX2 and AMPK.

A high-fat diet (HFD) causes obesity-associated morbidities involved in macroautophagy and chaperone-mediated autophagy (CMA). AMPK, the mediator of macroautophage, has been reported to be inactivated in HFD-caused renal injury. However, PAX2, the mediator for CMA, has not been reported in HFD-caused renal injury. Here we report that HFD-caused renal injury involved the inactivation of Pax2 and Ampk, and the activation of soluble epoxide hydrolase (sEH), in a murine model. Specifically, mice fed on an HFD for 2, 4, and 8 wk showed time-dependent renal injury, the significant decrease in renal Pax2 and Ampk at both mRNA and protein levels, and a significant increase in renal sEH at mRNA, protein, and molecular levels. Also, administration of an sEH inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea, significantly attenuated the HFD-caused renal injury, decreased renal sEH consistently at mRNA and protein levels, modified the renal levels of sEH-mediated epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) as expected, and increased renal Pax2 and Ampk at mRNA and/or protein levels. Furthermore, (PA) treatment caused significant increase in , and decrease in both Pax2 and Ampk in murine renal mesangial cells (mRMCs) time- and dose-dependently. Also, 14(15)-EET (a major substrate of sEH), but not its sEH-mediated metabolite 14,15-DHET, significantly reversed PA-induced increase in , and PA-induced decrease in Pax2 and Ampk. In addition, plasmid construction revealed that Pax2 may positively regulate Ampk transcriptionally in mRMCs. This study provides insights into and therapeutic target for the HFD-mediated renal injury.

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Inhibitory effect of 17β‑estradiol on triglyceride synthesis in skeletal muscle cells is dependent on ESR1 and not ESR2.

The present study aimed to investigate the inhibitory effects and the mechanisms underlying 17β‑estradiol (E2) effects on triglyceride synthesis and insulin resistance in skeletal muscle tissues and cells. Ovariectomy (OVX) was performed on 6‑month‑old female rats treated with or without E2. Subsequently, various serum biochemical markers were measured. Additionally, pathological alterations of the uterus, liver and skeletal muscle were analyzed, and the content of triglycerides (TG) in muscle was detected. Differentiated myotubes formed by C2C12 cells were treated with (PA) or pretreated with E2, estrogen receptor (ESR)\xa01 agonist propylpyrazoletriol (PPT) and ESR2 agonist diarylpropionitrile (DPN). Subsequently, the mRNA or protein expression levels of ESR1/2, peroxisome proliferator activated receptor α (PPARα), CD36 molecule (CD36), fatty synthase (FASN), perilipin 2 (PLIN2), phosphorylated acetyl‑CoA carboxylase α (p‑ACACA), p‑AKT serine/threonine kinase (p‑AKT) and p‑mitogen‑activated protein kinase\xa08 (p‑MAPK8) were analyzed in skeletal muscle or in C2C12 cells by reverse transcription‑semi‑quantitative polymerase chain reaction and western blotting. The present results suggested that treatment with E2 inhibited OVX‑induced gain, TG accumulation and insulin resistance. The protein or mRNA expression levels of ESR1, CD36, PPARα, p‑ACACA and p‑AKT were decreased, whereas the protein or mRNA expression levels of ESR2, PLIN2, FASN and p‑MAPK8 were increased in the OVX group. Of note, treatment with E2 restored the expression levels of the aforementioned factors. In C2C12 cells, treatment with E2 or PPT reversed the alterations induced by treatment with PA. In contrast, pretreatment with DPN did not influence the effect of PA. Collectively, E2 was able to interact with ESR1, thus activating the CD36‑PPARα pathway, decreasing the level of TG in the muscles and improving insulin resistance in skeletal muscles and C2C12 cells.

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Paeoniflorin Ameliorates Atherosclerosis by Suppressing TLR4-Mediated NF-κB Activation.

Paeoniflorin, a type of bioactive monoterpene glucoside in Paeoniae Radix, possesses anti-oxidative, anti-inflammatory and anti-hyperglycaemic properties. However, the underlying mechanism of paeoniflorin in treating atherosclerosis is unclear. A rat model of high-fat diet-induced atherosclerosis and (PA)-treated vascular smooth muscle cells (VSMCs) were used in this study. The serum concentrations of total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) were determined, and the results indicated that paeoniflorin remarkably lowered the levels of TC, TG and LDL-C induced by a high-fat diet. Histopathological results showed that paeoniflorin significantly improved the pathological changes in the aorta. In addition, paeoniflorin also maintained a normal gain speed. Subsequently, the effects of paeoniflorin on the production of inflammatory cytokines (IL-1β, IL-6 and TNF-α) were detected by qPCR and ELISA. The qPCR and ELISA results showed that paeoniflorin decreased the levels of these inflammatory cytokines. Moreover, the expression of TLR4 and its downstream pathway molecules was measured by Western blot. The results indicated that paeoniflorin significantly reduced the expression of TLR4 and MyD88 as well as the phosphorylation of IκBα and NF-κB p65. Taken together, these results suggested that paeoniflorin could alleviate atherosclerotic inflammation by inhibiting the TLR4/MyD88/NF-κB pathway. Therefore, paeoniflorin may be a potential therapy for atherosclerosis.

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Induction of Sphk1 activity in obese adipose tissue macrophages promotes survival.

During obesity, adipose tissue macrophages (ATM) are increased in concert with local inflammation and insulin resistance. Since the levels of sphingolipid (SLs) in adipose tissue (AT) are altered during obesity we investigated the potential impact of SLs on ATMs. For this, we first analyzed expression of SL metabolizing genes in ATMs isolated from obese mice. A marked induction of sphingosine kinase 1 (Sphk1) expression was observed in obese ATM when compared to lean ATM. This induction was observed in both MGL-ve (M1) and MGL1+ve (M2) macrophages from obese WAT. Next, RAW264.7 cells were exposed to excessive palmitate, resulting in a similar induction of Sphk1. This Sphk1 induction was also observed when cells were treated with chloroquine, a lysosomotropic amine impacting lysosome function. Simultaneous incubation of RAW cells with palmitate and the Sphk1 inhibitor SK1-I promoted cell death, suggesting a protective role of Sphk1 during lipotoxic conditions. Interestingly, a reduction of endoplasmic reticulum (ER) stress related genes was detected in obese ATM and was found to be associated with elevated Sphk1 expression. Altogether, our data suggest that lipid overload in ATM induces Sphk1, which promotes cell viability.

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Nutritional Value, Chemical Composition and Cytotoxic Properties of Common Purslane ( L.) in Relation to Harvesting Stage and Plant Part.

Purslane ( L.) is a widespread weed, which is highly appreciated for its high nutritional value with particular reference to the content in omega-3 fatty acids. In the present study, the nutritional value and chemical composition of purslane plants in relation to plant part and harvesting stage were evaluated. Plants were harvested at three growth stages (29, 43 and 52 days after sowing (DAS)), while the edible aerial parts were separated into stems and leaves. Leaves contained higher amounts of macronutrients than stems, especially at 52 DAS. α-tocopherol was the main isoform, which increased at 52 DAS, as well total tocopherols (values were in the ranges of 197-327 μg/100 g fresh (fw) and 302-481 μg/100 g fw, for α-tocopherol and total tocopherols, respectively). Glucose and fructose were the main free sugars in stems and leaves, respectively, whereas stems contained higher amounts of total sugars (values were ranged between 0.83 g and 1.28 g/100 g fw). Oxalic and total organic content was higher in leaves, especially at the last harvesting stage (52 DAS; 8.6 g and 30.3 g/100 g fw for oxalic and total organic acids, respectively). Regarding the fatty content, stems contained mainly (20.2-21.8%) and linoleic (23.02-27.11%), while leaves were abundant in α-linolenic (35.4-54.92%). Oleracein A and C were the major oleracein derivatives in leaves, regardless of the harvesting stage (values were in the ranges of 8.2-103.0 mg and 21.2-143 mg/100 g dried (dw) for oleraceins A and C, respectively). Cytotoxicity assays showed no hepatotoxicity, with GI values being higher than 400 μg/mL for all the harvesting stages and plant parts. In conclusion, early harvesting and the separation of plant parts could increase the nutritional value of the final product through increasing the content of valuable compounds, such as omega-3 fatty acids, phenolic compounds and oleracein derivatives, while at the same time, the contents of anti-nutritional compounds such as oxalic are reduced.

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Chemical evaluation of the Rhynchophorus ferrugineus larvae fed on different substrates as human food source.

We investigated the chemical composition of the weevil Rhynchophorus ferrugineus larvae, traditionally used as human food in Asia and known worldwide as one of the most significant pest for palm trees. Total fat content and fatty composition were analyzed using standard methodologies in (1) weevil larvae reared on apple fruit slices and wild specimens collected from attacked (2) Phoenix canariensis and (3) Syagrus romanzoffiana palm trees. Total fat content was extremely high in all the specimens (ranged between 57.62 and 60.03% based on dry ). Despite sharing the same prevalent fatty acids (myristic , , stearic , palmitoleic , oleic , α-linoleic , and α-linolenic ), fatty composition of the wild weevil larvae significantly differed from that of the specimens raised on apple fruit, due to the presence of other minor compounds. In general, a good balance between unsaturated fatty acids (∼53.68% of total fatty acids) and saturated fatty acids (∼43.41% of total fatty acids) and a low cholesterol content (74.61-152.32\u2009mg/kg based on dry matter) were detected in all the specimens. Conversely, the weevil larvae did not represent a good source of α-tocopherol (14.17-26.22\u2009mg/kg based on dry matter). The ability of the protein extracts obtained from the weevil larvae to inhibit in\xa0vitro the angiotensin-converting enzyme, the main enzyme involved in blood pressure regulation, was also investigated. To simulate gastrointestinal digestion process, protein extracts were hydrolyzed by the gastrointestinal enzymes. A significantly lower IC (0.588-0.623\u2009mg/ml) was measured in all the protein extracts after enzymatic hydrolysis versus the corresponding crude protein extracts (3.270-3.752\u2009mg/ml). Given that the weevil larvae are able to provide interesting benefits for human health, this study supports their use as human food not just in the native countries where they are traditionally consumed and farmed but also throughout the world.

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Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/lipid metabolism in type 2 diabetes mellitus. Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high-fat diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

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Screening of some lesser known tree-borne oilseed plants from North-East India for their oil content and major fatty components.

Forest of North-East India is rich in a variety of tree borne oilseeds and there were no serious efforts to exploit the less known oilseeds of the forest origin. Considering the vast plant wealth and with an objective to explore these untapped potentialities of oilseed resources of NE India, a research program was undertaken. The present communication is based on the investigation of seeds of 14 lesser known plant species for fats and oils. The screened plant species exhibited oil yield ranging from 4.71% to 48.43% on dry basis. Alseodaphne andersonii showed significant amount of tocols while Garcinia xanthochymus showed significant amount of phytosterol. Gas liquid chromatography analysis of methyl ester extracted fatty indicated dominant fraction of Oleic in 6 numbers of species, followed by Linoleic in 3 species, Lauric in 3 species, and in 2 species. The majority of the species (9 species) showed higher amount of unsaturated fatty content in the oils which suggested that the species may serve as valuable raw materials for vegetable oil which can be explored as substitute for industrial feasibility while restshowed higher amount of saturated fatty acids which suggested that these species may serve as important industrial by-products.Copyright © 2017 Elsevier Ltd. All rights reserved.

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Inhibition of galectin-3 ameliorates the consequences of cardiac lipotoxicity in a rat model of diet-induced obesity.

Obesity is accompanied by metabolic alterations characterized by insulin resistance and cardiac lipotoxicity. Galectin-3 (Gal-3) induces cardiac inflammation and fibrosis in the context of obesity; however, its role in the metabolic consequences of obesity is not totally established. We have investigated the potential role of Gal-3 in the cardiac metabolic disturbances associated with obesity. In addition, we have explored whether this participation is, at least partially, acting on mitochondrial damage. Gal-3 inhibition in rats that were fed a high-fat diet (HFD) for 6\u2005weeks with modified citrus pectin (MCP; 100\u2005mg/kg/day) attenuated the increase in cardiac levels of total triglyceride (TG). MCP treatment also prevented the increase in cardiac protein levels of carnitine palmitoyl transferase IA, mitofusin 1, and mitochondrial complexes I and II, reactive oxygen species accumulation and decrease in those of complex V but did not affect the reduction in F-fluorodeoxyglucose uptake observed in HFD rats. The exposure of cardiac myoblasts (H9c2) to increased the rate of respiration, mainly due to an increase in the proton leak, glycolysis, oxidative stress, β-oxidation and reduced mitochondrial membrane potential. Inhibition of Gal-3 activity was unable to affect these changes. Our findings indicate that Gal-3 inhibition attenuates some of the consequences of cardiac lipotoxicity induced by a HFD since it reduced TG and lysophosphatidyl choline (LPC) levels. These reductions were accompanied by amelioration of the mitochondrial damage observed in HFD rats, although no improvement was observed regarding insulin resistance. These findings increase the interest for Gal-3 as a potential new target for therapeutic intervention to prevent obesity-associated cardiac lipotoxicity and subsequent mitochondrial dysfunction© 2018. Published by The Company of Biologists Ltd.

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METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, insulin resistance, and diabetes. Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

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Yield, Antioxidant Components, Oil Content, and Composition of Onion Seeds Are Influenced by Planting Time and Density.

Research was carried out on onion landrace (Ramata di Montoro) for seed production in southern Italy, with the aim to evaluate the effects on yield and quality of four bulb planting times in factorial combination with four densities, using a split plot design with three replicates. The number of flower stalks per plant, their height and diameter, and the inflorescence diameter decreased with the bulb planting delay and density increase. The highest plant leaf area and LAI (leaf area index), seed yield, number, and mean were recorded with the earliest planting time, with the lowest bulb density eliciting the highest plant leaf area but the lowest LAI and seed yield per hectare. The ratio between seeds and inflorescence , and seed germinability, decreased with the planting delay and density increase. Seed oil, protein, and antioxidant content (polyphenols and selenium) were highest with the last crop cycle. The polyunsaturated fatty acids, predominant in oil, increased with planting time delay, whereas the monounsaturated fatty acids decreased. Linoleic, oleic, and prevailed among polyunsaturated, monounsaturated, and saturated fatty acids, respectively. Planting from 20 December to 10 January with 3.3 cold-stored bulbs per m was the most effective combination in terms of seed yield per hectare, whereas seed oil content and quality were the best, with the last crop cycle starting on 21 February, independent of bulb density.

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Optimization growth of Spirulina (Arthrospira) platensis in photobioreactor under varied nitrogen concentration for maximized biomass, carotenoids and lipid contents.

Spirulina (Arthrospira) platensis (SP) microalgae were cultured in Zarrouk medium (ZM), containing three nitrogen concentrations (N-limited, N-optimal and N-rich medium) in ten liter-photo-bioreactor (10 L PBR) for 15-days, in order to study changes in lipid compounds (total carotenoids and total lipids and their effect on fatty profile). Based on US patent, the yield of bioactive compounds (such as gamma-linolenic GLA, C18:3) extracted from microalgae biomass, mainly depends on the extraction processes (1). GLA has much attention in respect of its therapeutic properties such as its ability to decrease blood cholesterol levels.The impact of the addition of N in cultures of S. platensis in terms of growth, biomasses and induced lipid compounds (total carotenoids and total lipid contents and its fatty profile) as well as the sonication (SON) and microwave (MIC) process as aiding techniques for lipid extraction compared with a cold condition (COL) were examined. GC/MS method was used to determine the fatty profile of lipid extract of SP cultures.In all S. platensis tested culture, the SP was growing successfully, with varied degrees. In N-rich media, the highest cell growth rate and biomass yield were obtained compared with that recorded in other cultures. Under an N-limited condition, SP had a higher total carotenoids (TCAR, 45.54 mg/g dw) and total lipid contents (TL, 29.51%± 1.92 g/100g dw) compared with that recorded either in N-rich (11.2 mg/g dw) or in N-optimal (6.23 mg/g dw) cultures. Thus, SP copes with the N -stress by altering the metabolic pathways toward inducing lipid biosynthesis. To maximize the TL and TCAR extraction yields, from N-limited cultures, a set of operating process was applied including the sonication (SON) and microwave (MIC), which were used as aiding techniques for lipid extraction compared with the cold condition (COL) techniques. The results showed that the extraction efficiency of the S. platensis TL increased in the following order: MIC (29.51%± 1.92) > SON (25.46% ± 1.65> COL (20.43% ±1.43). In comparative study for its fatty profiles (FAPs) among all SP cultures, lipids were analyzed by GC/MS. The predominant fatty acids (>10%, of total FA) were found to be myristic (C14:0, MA), (C16:0, PA) and oleic (C18:1).The study concluded that N-limited condition was found to be a strong influence on biomass dry and lipid contents and total carotenoids in SP cells compared to either N-rich or N-optimal conditions. The use of sonication and the microwave techniques lead to great increase in the extraction of lipid contents and in high amount polyunsaturated fatty acids (PUFAs) in N-limited cultures, in particular the omega-6 (ω6) and omega-3 (ω3) of the essential C18 fatty acids. It seems that the SP rich in lipid content with a high amount of GLC produced under nitrogen limitation in PBR conditions can be used as a food additive or as a nutritional supplement.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

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Optimisation, experimental validation and thermodynamic study of the sequential oil extraction and biodiesel production processes from seeds of Sterculia foetida.

Non-edible seeds are not used in any commercial applications, which implies that they can be used for biofuel applications. The present study aimed to maximise the process conditions for oil extraction and sterculia biodiesel production from Sterculia foetida (poon oil). GC-MS identified the methyl esters of sterculia oil as sterculic (32%), (15.88%), oleic (10.00%), linoleic (9.95%) and malvalic (9%). Response surface methodology (RSM) based parametric optimisation of oil extraction was carried out by choosing process variables such as sample , volume of solvent to seed ratio and time. The optimum sample amount of 7.5 g and the volume of solvent to seed ratio of 40 mL/g resulted in a maximum oil yield of 45.27% at 3 h. The results were statistically significant (P < 0.05) with a regression coefficient (R) of 0.9988. Furthermore, the artificial neural network (ANN) resulted in an R value greater than 0.9, which validates the RSM. Conventional optimisation of the temperature (55 °C), feedstock to methanol ratio (1:12), catalyst proportion (1.5%) and transesterification reaction time (60 min) yield 90.87% biodiesel production. The physicochemical characteristics of oil and biodiesel complied with the requirements of the ASTM standards. The rate constant and thermodynamic variables at the optimum temperature (333 K) were calculated from the experimental data. The activation energy (E), activation enthalpy in transition state theory (ΔH), activation entropy in transition state theory (ΔS) and Gibbs free energy in transition state theory (ΔG) were 37.91 kJ mol, 35.14 kJ mol, - 239.58 J mol K and 79.81 kJ mol respectively. Graphical abstract.

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Adipocyte-Derived Exosomal MiR-27a Induces Insulin Resistance in Skeletal Muscle Through Repression of PPARγ.

The mechanism by which adipocyte-derived endocrine factors promote insulin resistance in skeletal muscle are not fully understood. MiR-27a is highly expressed in sera of obese individuals with prediabetes and T2DM, and mainly derived by adipose tissues. Thus, miR-27a secreted into circulation by adipose tissue may regulate insulin resistance in skeletal muscle. The association between miR-27a and insulin resistance in skeletal muscle was determined in obese children, high-fat diet-induced miR-27a knockdown obese mice, db/db mice and C2C12 cells overexpressing miR-27a. The crosstalk mediated by exosomal miR-27a between adipose tissue and skeletal muscle was determined in C2C12 cells incubated with conditioned medium prepared from palmitate-treated 3T3-L1 adipocytes. We showed that serum miR-27a level correlated positively with obesity and insulin resistance in obese children, and that elevated serum miR-27a levels correlated with insulin resistance in leptin receptor-deficient db/db mice, and with obesity and insulin resistance in high-fat diet-fed C57BL/6J mice. MiR-27a released from adipocytes of high-fat diet-fed C57BL/6J mice was associated with triglyceride accumulation. MiR-27a derived from these adipocytes induced insulin resistance in C2C12 skeletal muscle cells through miR-27a-mediated repression of PPARγ and its downstream genes involved in the development of obesity. These results identify a novel crosstalk signaling pathway between adipose tissue and skeletal muscle in the development of insulin resistance, and indicate that adipose tissue-derived miR-27a may play a key role in the development of obesity-triggered insulin resistance in skeletal muscle.

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Gut Microbiota and Metabolome Response of Seed Oil on Metabolism Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut microbiota dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.

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PINK1-Parkin alleviates metabolic stress induced by obesity in adipose tissue and in 3T3-L1 preadipocytes.

Mitochondria play an important role in cellular metabolism and are closely related with metabolic stress. Recently, several studies have shown that mitophagy mediated by PTEN-induced putative kinase 1 (PINK1) and Parkin may play a critical role in clearing the damaged mitochondria and maintaining the overall balance of intracellular mitochondria in quality and quantity. A previous study showed that PINK1 and Parkin were overexpressed in adipose tissue in obese subjects. However, it is still unclear whether a direct relationship exists between obesity and mitophagy. In this study, we created a high-fat-diet (HFD)-induced obese mouse model and examined the expression of PINK1 and Parkin in adipose tissue using western blot and real-time quantitative PCR. After we confirmed that there is an interesting difference between regular-chow-fed mice and HFD-induced obese mice in the expression of PINK1 and Parkin in\xa0vivo, we further tested the expression of PINK1 and Parkin in 3T3-L1 preadipocytes in\xa0vitro by treating cells with (PA) to induce metabolic stress. To better understand the role of PINK1 and Parkin in metabolic stress, 3T3-L1 preadipocytes were transfected with small interfering RNA (siRNA) of PINK1 and Parkin followed by PA treatment. Our results showed that under lower concentrations of PA, PINK1 and Parkin can be activated and play a protective role in resisting the harmful effects of PA, including protecting the mitochondrial function and resisting cellular death, while under higher concentrations of PA, the expression of PINK1 and Parkin can be inhibited. These results suggest that PINK1-Parkin can protect mitochondrial function against metabolic stress induced by obesity or PA to a certain degree.Copyright © 2018 Elsevier Inc. All rights reserved.

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ALDH2 protects against high fat diet-induced obesity cardiomyopathy and defective autophagy: role of CaM kinase II, histone H3K9 methyltransferase SUV39H, Sirt1, and PGC-1α deacetylation.

Uncorrected obesity contributes to cardiac remodeling and contractile dysfunction although the underlying mechanism remains poorly understood. Mitochondrial aldehyde dehydrogenase (ALDH2) is a mitochondrial enzyme with some promises in a number of cardiovascular diseases. This study was designed to evaluate the impact of ALDH2 on cardiac remodeling and contractile property in high fat diet-induced obesity.Wild-type (WT) and ALDH2 transgenic mice were fed low (10% calorie from fat) or high (45% calorie from fat) fat diet for 5 months prior to the assessment of cardiac geometry and function using echocardiography, IonOptix system, Lectin, and Masson Trichrome staining. Western blot analysis was employed to evaluate autophagy, CaM kinase II, PGC-1α, histone H3K9 methyltransferase SUV39H, and Sirt-1.Our data revealed that high fat diet intake promoted gain, cardiac remodeling (hypertrophy and interstitial fibrosis, p\u2009<\u20090.0001) and contractile dysfunction (reduced fractional shortening (p\u2009<\u20090.0001), cardiomyocyte function (p\u2009<\u20090.0001), and intracellular Ca handling (p\u2009=\u20090.0346)), mitochondrial injury (elevated O levels, suppressed PGC-1α, and enhanced PGC-1α acetylation, p\u2009<\u20090.0001), elevated SUV39H, suppressed Sirt1, autophagy and phosphorylation of AMPK and CaM kinase II, the effects of which were negated by ALDH2 (p\u2009≤\u20090.0162). In vitro incubation of the ALDH2 activator Alda-1 rescued against -induced changes in cardiomyocyte function, the effect of which was nullified by the Sirt-1 inhibitor nicotinamide and the CaM kinase II inhibitor KN-93 (p\u2009<\u20090.0001). The SUV39H inhibitor chaetocin mimicked Alda-1-induced protection again (p\u2009<\u20090.0001). Examination in overweight human revealed an inverse correlation between diastolic cardiac function and ALDH2 gene mutation (p\u2009<\u20090.05).Taken together, these data suggest that ALDH2 serves as an indispensable factor against cardiac anomalies in diet-induced obesity through a mechanism related to autophagy regulation and facilitation of the SUV39H-Sirt1-dependent PGC-1α deacetylation.

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Apigenin Ameliorates the Obesity-Induced Skeletal Muscle Atrophy by Attenuating Mitochondrial Dysfunction in the Muscle of Obese Mice.

It was investigated whether apigenin (AP) protected against skeletal muscle atrophy induced by obesity.Mice were fed a high-fat diet (HFD) for 9 weeks to induce obesity, and then were assigned to two groups; the HFD group received a high-fat diet, and the HFD+AP group received a 0.1% AP-containing HFD. After additional feeding of the experimental diet for 8 weeks, mice in the HFD group were highly obese compared with the mice in the standard diet fed mice group. The mice in the AP-treated group showed less fat pad accumulation and less inflammatory cytokines without reduction. The of skeletal muscle in the AP group tended to increase as compared with that of the HFD group. Furthermore, AP reduced the expression of atrophic genes, including MuRF1 and Atrogin-1, but increased the exercise capacity. The mitochondrial function and mitochondrial biogenesis were enhanced by AP. In cultured C2C12 cells, AP also suppressed -induced muscle atrophy and mitochondrial dysfunction. In addition, AP activated AMP-activated protein kinase (AMPK) in the C2C12 and the muscle of HFD-induced obese mice.The results suggested that AP ameliorated the obesity-induced skeletal muscle atrophy by attenuating mitochondrial dysfunction.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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The nutritive value of marula (Sclerocarya birrea) seed cake for broiler chickens: nutritional composition, performance, carcass characteristics and oxidative and mycotoxin status.

This study was aimed at investigating the nutritive value of marula seed cake (MSC) as an alternative protein source for broilers. In a completely randomised design involving six replicate pens of five chickens assigned to each of five treatments, gain (BWG), feed intake (FI), feed conversion efficiency (FCE) and carcass characteristics were measured in an experiment in which 150 28-day-old broilers were fed maize-based diets containing, respectively, 0, 5, 10, 15 and 20% MSC at finisher phase. The results showed MSC to be remarkably high in CP (470.0\xa0g/kg DM) and EE (343.5\xa0g/kg DM), with moderate CF (58.2\xa0g/kg DM), ash (54.3\xa0g/kg DM), Ca (1.1\xa0g/kg DM) and P (11.0\xa0g/kg DM). Whilst very poor in lysine, MSC was found to be rich in methionine, cyst(e)ine, arginine and glutamic ; it also contains good levels of valine, glycine, threonine, isoleucine, leucine, histidine, phenylalanine, serine, proline and alanine. Also, it contained 85.24% oleic (OA), 9.65% and 5.11% stearic acids, with a high peroxide value and low levels of mycotoxins deoxynivalenol (DON) and T-2 toxin. BWG, FI and FCE of broiler chickens significantly decreased (P\xa0<\xa00.001) as the dietary level of MSC increased. Further, dietary MSC significantly decreased bird live at slaughter (P\xa0<\xa00.001), plucked (P\xa0<\xa00.001), dressed (P\xa0<\xa00.001) and of the liver (P\xa0<\xa00.001) and neck (P\xa0<\xa00.05). The results therefore demonstrate MSC to be a good source of CP, fat, Ca, P, amino acids (except lysine) and OA that can replace soya bean meal (SBM) in broiler diets. However, its use is currently limited by lipid peroxidation and presence of mycotoxins.

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Metabolomic elucidation of the effects of media and carbon sources on fatty production by Yarrowia lipolytica.

Lipid production by oleaginous Yarrowia lipolytica depends highly on culture environments, such as carbon sources, carbon/nitrogen (C/N) ratios, types of media, and cellular growth phases. In this study, the effects of media and carbon sources on lipid and metabolite production were investigated by profiling fatty acids and intracellular metabolites of Y. lipolytica grown in various media. The highest total fatty yield 114.04\u202f±\u202f6.23\u202fmg/g dry cell was achieved by Y. lipolytica grown in minimal medium with glycerol (SCG) in the exponential phase. The high lipid production by Y. lipolytica in SCG was presumed to be due to the higher C/N ratio in SCG than in the complex media. Moreover, glycerol promoted lipid production better than glucose in both complex and minimal media because glycerol can easily incorporate into the core of triglycerides. Metabolite profiling revealed that levels of long-chain fatty acids, such as stearic , , and arachidic , increased in SCG medium. Meanwhile, in complex media supplemented with either glucose or glycerol, levels of amino acids, such as cysteine, methionine, and glycine, highly increased. This metabolomic approach could be applied to modulate the global metabolic network of Y. lipolytica for producing lipids and other valuable products.Copyright © 2018 Elsevier B.V. All rights reserved.

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Hydroxystearic Acids Activate GPR40, Which Is Involved in Their Beneficial Effects on Glucose Homeostasis.

hydroxystearic acids (PAHSAs) are endogenous lipids with anti-diabetic and anti-inflammatory effects. PAHSA levels are reduced in serum and adipose tissue of insulin-resistant people and high-fat diet (HFD)-fed mice. Here, we investigated whether chronic PAHSA treatment enhances insulin sensitivity and which receptors mediate PAHSA effects. Chronic PAHSA administration in chow- and HFD-fed mice raises serum and tissue PAHSA levels ∼1.4- to 3-fold. This improves insulin sensitivity and glucose tolerance without altering . PAHSA administration in chow-fed, but not HFD-fed, mice augments insulin and glucagon-like peptide (GLP-1) secretion. PAHSAs are selective agonists for GPR40, increasing Ca flux, but not intracellular cyclic AMP. Blocking GPR40 reverses improvements in glucose tolerance and insulin sensitivity in PAHSA-treated chow- and HFD-fed mice and directly inhibits PAHSA augmentation of glucose-stimulated insulin secretion in human islets. In contrast, GLP-1 receptor blockade in PAHSA-treated chow-fed mice reduces PAHSA effects on glucose tolerance, but not on insulin sensitivity. Thus, PAHSAs activate GPR40, which is involved in their beneficial metabolic effects.Copyright © 2018 Elsevier Inc. All rights reserved.

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Effects of heat-treated hempseed supplementation on performance, egg quality, sensory evaluation and antioxidant activity of laying hens.

1. This study was conducted to determine the effects of raw and heat-treated hempseed (HHS, Cannabis sativa L.) on performance, egg quality and antioxidant activity in laying hens. 2. A total of 108 laying hens, aged 36\xa0weeks, were divided into three treatment groups with 12 replicates and each replicate contained three laying hens. The treatments were as follows: (1) Control (no hempseed), (2) 15% raw hempseed (RHS) in diet and (3) 15% HHS in the diet. Experiments lasted for 12\xa0weeks. 3. Feed intake of the RHS group was lower than those of the control and HHS groups. Egg , egg mass, shell , shell surface area and shell thickness of the HHS group were significantly (P <\xa00.05) higher than that of the RHS group. Roche (DSM) colour fan values of the RHS group were higher than that of the HHS group (P <\xa00.01). 4. , palmitoleic and oleic acids of egg yolk were significantly (P <\xa00.05) decreased in the RHS and HHS groups; however, linoleic, α-linolenic and docosahexaenoic acids (DHA) of egg yolk increased (P <\xa00.05) for both treatment diets compared to the control group. 5. Both RHS and HHS supplementation to layer diets did not influence malondialdehyde (MDA) and superoxide dismutase (SOD) activity and blood lipid profile. 6. It was concluded that HHS was superior in improving the egg quality of laying hens as compared to the RHS.

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Physicochemical Quality, Fatty Composition, and Sensory Analysis of Nellore Steers Meat Fed with Inclusion of Condensed Tannin in the Diet.

This study was conducted to test the effect of dietary tannin on the fatty profile and sensory attributes of meat from Nellore steers. Thirty-two Nellore bull male were distributed in a completely randomized design and fed diets with condensed tannin extract as follows: 0, 10, 30, and 50\xa0g/kg total DM basis. The physicochemical composition of the meat, lipid oxidation, fatty profile, flavor, tenderness, and overall acceptance were evaluated. There was a linear decrease (P\xa0≤\xa00.05) on lipid content, tenderness, cooking loss, myristic, , and oleic acids in meat as tannin increased in the diets. The total saturated and monounsaturated fatty acids, the atherogenicity index decreased. However, a linear increase (P\xa0≤\xa00.05) was observed for linoleic, linolenic, arachidonic, eicosapentaenoic, and docosapentaenoic acids. The physicochemical characteristic of the meat, such as moisture, ash, and protein contents, water retention capacity, final pH, Warner-Bratzler shear force, collagen, and color indexes (lightness, redness, yellowness, and chrome) did not change with dietary tannin. Also, CLA, n-6:n-3\xa0ratio, Δ -desaturase, and elongase activity were not different among diets. In conclusion, condensed tannin linearly increases unsaturated fatty acids and decreases the atherogenicity index of meat; thus, it can be recommended at the highest level (50\xa0g/kg DM) in the diet of Nellore steers.Agriculture byproducts plays an important part in the diet of ruminant animals and consequently on food chain and has implications for the composition and quality of the livestock products (milk, meat, and eggs) that people consume. Feeding tannin to steers increases the amount of unsaturated fatty acids and meat tenderness, with a concomitant reduction on saturated fatty acids and the atherogenicity index in meat. Thus, we recommend adding tannin to steer diets to reduce the risk factors for cardiovascular diseases in red meat for human consumption.© 2018 Institute of Food Technologists®.

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Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration.

Obesity may promote intervertebral disc degeneration (IDD) by non-mechanical means, by influencing levels of free fatty acids which could impair cell metabolism. This study aims to establish metabolic factors in obesity-related IDD independent of mechanical loading. In clinical study, we retrospectively reviewed 128 volunteers (73 males, 55 females, aged 29-88 years) and compared their grades of disk degeneration with obesity-related factors such as , BMI, and serum lipid levels. Clinically, the IDD group showed increased age, BMI and serum triglyceride. Triglyceride was a significant risk factor for IDD even after correction for BMI and age (P = 0.007). In obesity animal model, rats were fed a high-fat diet (HFD) in order to study its effects on disk metabolism and apoptosis. HFD rats had significantly higher serum levels of lipids, including triglyceride and non-esterified fatty , and showed significantly decreased markers of anabolism, increased catabolism and apoptosis in disk. Finally, rat nucleus pulposus (NP) cells were stimulated with a fatty (, PA) to gauge its effects on cell metabolism and apoptosis. Cell culture studies showed that NP cells exposed to PA showed increased apoptosis for activation of caspase 3, 7, 9, and PARP, which was primarily via the MAPK signal pathway, especially ERK pathway. In conclusion, hypertriglyceridemia can lead to IDD, independently of age and BMI. Hypertriglyceridemia appears to mediate disk cell apoptosis and matrix catabolism primarily via the ERK pathway.Copyright © 2019 Zhang, Chen, Huang, Wang, Shan, Liu, Chen, Li, Fan and Zhao.

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Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue , glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

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MDD-Palm: Identification of protein S-palmitoylation sites with substrate motifs based on maximal dependence decomposition.

S-palmitoylation, the covalent attachment of 16-carbon acids to a cysteine residue via a thioester linkage, is an important reversible lipid modification that plays a regulatory role in a variety of physiological and biological processes. As the number of experimentally identified S-palmitoylated peptides increases, it is imperative to investigate substrate motifs to facilitate the study of protein S-palmitoylation. Based on 710 non-homologous S-palmitoylation sites obtained from published databases and the literature, we carried out a bioinformatics investigation of S-palmitoylation sites based on amino composition. Two Sample Logo indicates that positively charged and polar amino acids surrounding S-palmitoylated sites may be associated with the substrate site specificity of protein S-palmitoylation. Additionally, maximal dependence decomposition (MDD) was applied to explore the motif signatures of S-palmitoylation sites by categorizing a large-scale dataset into subgroups with statistically significant conservation of amino acids. Single features such as amino composition (AAC), amino pair composition (AAPC), position specific scoring matrix (PSSM), position matrix (PWM), amino substitution matrix (BLOSUM62), and accessible surface area (ASA) were considered, along with the effectiveness of incorporating MDD-identified substrate motifs into a two-layered prediction model. Evaluation by five-fold cross-validation showed that a hybrid of AAC and PSSM performs best at discriminating between S-palmitoylation and non-S-palmitoylation sites, according to the support vector machine (SVM). The two-layered SVM model integrating MDD-identified substrate motifs performed well, with a sensitivity of 0.79, specificity of 0.80, accuracy of 0.80, and Matthews Correlation Coefficient (MCC) value of 0.45. Using an independent testing dataset (613 S-palmitoylated and 5412 non-S-palmitoylated sites) obtained from the literature, we demonstrated that the two-layered SVM model could outperform other prediction tools, yielding a balanced sensitivity and specificity of 0.690 and 0.694, respectively. This two-layered SVM model has been implemented as a web-based system (MDD-Palm), which is now freely available at http://csb.cse.yzu.edu.tw/MDDPalm/.

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Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

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Effects of increased myocardial tissue concentration of myristic, and palmitoleic acids on the course of cardiac atrophy of the failing heart unloaded by heterotopic transplantation.

The present experiments were performed to evaluate if increased heart tissue concentration of fatty acids, specifically myristic, and palmitoleic acids that are believed to promote physiological heart growth, can attenuate the progression of unloading-induced cardiac atrophy in rats with healthy and failing hearts. Heterotopic abdominal heart transplantation (HT(x)) was used as a model for heart unloading. Cardiac atrophy was assessed from the ratio of the native- to-transplanted heart (HW). The degree of cardiac atrophy after HT(x) was determined on days 7, 14, 21 and 28 after HT(x) in recipients of either healthy or failing hearts. HT(x) of healthy hearts resulted in 23+/-3, 46+/-3, 48+/-4 and 46+/-4 % HW loss at the four time-points. HT(x) of the failing heart resulted in even greater HW losses, of 46+/-4, 58+/-3, 66+/-2 and 68+/-4 %, respectively (P<0.05). Activation of "fetal gene cardiac program" (e.g. beta myosin heavy chain gene expression) and "genes reflecting cardiac remodeling" (e.g. atrial natriuretic peptide gene expression) after HT(x) was greater in failing than in healthy hearts (P<0.05 each time). Exposure to isocaloric high sugar diet caused significant increases in fatty concentrations in healthy and in failing hearts. However, these increases were not associated with any change in the course of cardiac atrophy, similarly in healthy and post-HT(x) failing hearts. We conclude that increasing heart tissue concentrations of the fatty acids allegedly involved in heart growth does not attenuate the unloading-induced cardiac atrophy.

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Altering the ratio of dietary , stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, energy partitioning, and production responses of dairy cows.

The objective of this study was to evaluate the effects of varying the ratio of dietary (C16:0), stearic (C18:0), and oleic (cis-9 C18:1) acids in basal diets containing soyhulls or whole cottonseed on nutrient digestibility, energy partitioning, and production response of lactating dairy cows. Twenty-four mid-lactation multiparous Holstein cows were used in a split-plot Latin square design. Cows were allocated to a main plot receiving either a basal diet with soyhulls (SH, n = 12) or a basal diet with whole cottonseed (CS, n = 12) that was fed throughout the experiment. Within each plot a 4 × 4 Latin square arrangement of treatments was used in 4 consecutive 21-d periods. Treatments were (1) control (CON; no supplemental fat), (2) high C16:0 supplement [PA; fatty (FA) supplement blend provided ∼80% C16:0], (3) C16:0 and C18:0 supplement (PA+SA; FA supplement blend provided ∼40% C16:0 + ∼40% C18:0), and (4) C16:0 and cis-9 C18:1 supplement (PA+OA; FA supplement blend provided ∼45% C16:0 + ∼35% cis-9 C18:1). Interactions between basal diets and FA treatments were observed for dry matter intake (DMI) and milk yield. Among the SH diets, PA and PA+SA increased DMI compared with CON and PA+OA treatments, whereas in the CS diets PA+OA decreased DMI compared with CON. The PA, PA+SA, and PA+OA treatments increased milk yield compared with CON in the SH diets. The CS diets increased milk fat yield compared with the SH diets due to the greater yield of de novo and preformed milk FA. The PA treatment increased milk fat yield compared with CON, PA+SA, and PA+OA due to the greater yield of mixed-source (16-carbon) milk FA. The PA treatment increased 3.5% fat-corrected milk compared with CON and tended to increase it compared with PA+SA and PA+OA. The CS diets increased (BW) change compared with the SH diets. Additionally, PA+OA tended to increase BW change compared with CON and PA and increased it in comparison with PA+SA. The PA and PA+OA treatments increased dry matter and neutral detergent fiber digestibility compared with PA+SA and tended to increase them compared with CON. The PA+SA treatment reduced 16-carbon, 18-carbon, and total FA digestibility compared with the other treatments. The CS diets increased energy partitioning toward reserves compared with the SH diets. The PA treatment increased energy partitioning toward milk compared with CON and PA+OA and tended to increase it compared with PA+SA. In contrast, PA+OA increased energy partitioned to reserves compared with PA and PA+SA and tended to increase it compared with CON. In conclusion, milk yield responses to different combinations of FA were affected by the addition of whole cottonseed in the diet. Among the combinations of C16:0, C18:0, and cis-9 C18:1 evaluated, fat supplements with more C16:0 increased energy output in milk, whereas fat supplements with more cis-9 C18:1 increased energy storage in BW. The combination of C16:0 and C18:0 reduced nutrient digestibility, which most likely explains the lower performance observed compared with other treatments.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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Low molecular fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic fatty-liver disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common liver disease globally. Low molecular fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in diabetes-induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the liver of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for obesity/diabetes-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

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Chemical composition, in vitro gas production, methane production and fatty profile of canola silage (Brassica napus) with four levels of molasses.

The objective of this study was to investigate the effect of four levels of molasses on chemical composition, in vitro digestibility, methane production and fatty profile of canola silages. A canola (Brassica napus var. Monty) crop was established in a small-scale agricultural farm and harvested 148\xa0days after sowing. Four levels of molasses were tested with respect to the fresh (1.5\xa0kg); these were 1% (CS-1), 2% (CS-2), 3% (CS-3) and 4% (CS-4) molasses, and 0% molasses (CS-0) was included as a control. A total of 45 microsilages were prepared using PVC pipes (4\xa0in. of diameter × 20\xa0cm of length), and the forage was compressed using a manual press. The effects of control and treatments were tested using the general linear model Y\u2009=\u2009μ\u2009+\u2009T\u2009+\u2009E. The linolenic (C18:3n3), (C16:0) and linoleic methyl ester (C18:2n6c) accounted for 30%, 21% and 10.5% of total fatty acids, respectively; the fermentation parameters and in vitro methane production were not affected (P\u2009>\u20090.05) by treatments; in vitro digestibility decreased significantly (P\u2009<\u20090.05) as the level of molasses increased. It was concluded that CS-4 improved the DM content by 9% and showed high content of linolenic methyl ester. The gross energy of canola silages could favour the oleic methyl ester.

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Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut microbiota, and hepatic metabolism.

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Saturated Fatty Increases Lung Macrophages and Augments House Dust Mite-Induced Airway Inflammation in Mice Fed with High-Fat Diet.

Obesity is one of the phenotypes of severe asthma, which is considered to be a heterogeneous syndrome; however, its interaction with airway inflammation is not fully understood. The aim of this study was to clarify the role of saturated fatty acids in augmenting airway inflammation induced by house dust mite (HDM) in obesity. Subjects were Balb/c mice fed a high-fat diet (HFD) for 10\xa0weeks, followed by sensitization and exposure to HDM. Subjects were also administered (PA) for 4\xa0weeks with concurrent sensitization and exposure to HDM. Airway inflammation was assessed by quantifying the amount of inflammatory cells in bronchoalveolar lavage (BAL) and airway resistance was measured. In vitro, lipopolysaccharide (LPS)-primed macrophages were stimulated by PA. The amount of monocyte chemoattractant protein-1 (MCP-1), interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) was examined in the supernatant. Compared to normal chow mice, HFD mice underwent significant increases in ; increases in number of lung macrophages, including circulating monocytes and alveolar macrophages; and increases in bronchoalveolar lavage fluid (BALF) total cell count, including neutrophils but not eosinophils, after HDM sensitization and exposure. In vitro, PA induced MCP-1 and augmented LPS-primed production of IL-1β and TNF-α in macrophages. Among HDM mice that were administered PA, there was an increase BALF total cell count, including neutrophils but not eosinophils, compared to vehicle mice. In conclusion, saturated fatty increased the number of lung macrophages and augmented HDM-induced neutrophilic airway inflammation in a HFD mouse model.

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Palmitate-Induced Insulin Hypersecretion and Later Secretory Decline Associated with Changes in Protein Expression Patterns in Human Pancreatic Islets.

In obese children with high circulating concentrations of free fatty palmitate, we have observed that insulin levels at fasting and in response to a glucose challenge were several times higher than in obese children with low concentrations of the fatty as well as in lean controls. Declining and even insufficient insulin levels were observed in obese adolescents with high levels of the fatty . In isolated human islets exposed to palmitate we have observed insulin hypersecretion after 2 days exposure. In contrast, insulin secretion from the islets was reduced after 7 days culture in the presence of the fatty . This study aims\xa0at identifying islet-related biological events potentially linked with the observed insulin hypersecretion and later secretory decline in these obese children and adolescents using the islet model. We analyzed protein expression data obtained from human islets exposed to elevated palmitate levels for 2 and 7 days by an improved methodology for statistical analysis of differentially expressed proteins. Protein profiling of islet samples by liquid chromatography-tandem mass spectrometry identified 115 differentially expressed proteins (DEPs). Several DEPs including sorcin were associated with increased glucose-stimulated insulin secretion\xa0in islets after 2 days of exposure to palmitate. Similarly, several metabolic pathways including altered protein degradation, increased autophagy, altered redox condition, and hampered insulin processing were coupled to the functional impairment of islets after 7 days of culture in the presence of palmitate. Such biological events, once validated in the islets, may give rise to novel treatment strategies aiming at normalizing insulin levels in obese children with high palmitate levels, which may reduce or even prevent obesity-related type 2 diabetes mellitus.

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Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

Obesity increases the risk of developing diabetes mellitus. Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in diabetes with obesity. Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II diabetes. Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling pathways was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of diabetes with obesity that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for diabetes with obesity.

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Antarctic thraustochytrids: Producers of long-chain omega-3 polyunsaturated fatty acids.

Thraustochytrids have been isolated from different aquatic systems; however, few studies have reported their occurrence in Antarctica. In this study, 13 strains close to strains belonging to the genera Oblongichytrium, Thraustochytrium, and Aurantiochytrium were isolated from seawater samples collected near the Antarctic Base Professor Julio Escudero (S 62°12\'57\' E 58°57\'35″). Docosahexaenoic (DHA) was found in the total lipids of all the isolates; DHA content of the biomass (dry ) varied between 3.3 and 33\xa0mg/g under the growth conditions for isolation. Five of the Antarctic thraustochytrids were able to accumulate lipids at levels higher than 20% w/w. Two strains, RT2316-7 and RT2316-13, were selected to test the effect of the incubation temperature (at 5°C for 14\xa0days and at 15°C for 5\xa0days). Incubation temperature had little effect on the lipid content and biomass yield; however, its effect on the fatty composition was significant (p\xa0<\xa0.05). The low incubation temperature favored the accumulation of eicosapentaenoic (EPA), and stearic in the total lipids of RT2316-7. Percentage of EPA, DHA and the omega-6 fatty dihomo-γ-linolenic of total fatty acids of RT2316-13 was higher at the low incubation temperature. RT2316-13 accumulated the highest lipid content (30.0\xa0±\xa00.5%) with a carbon to nitrogen mass ratio equal to 16.9. On the contrary, lipid accumulation in RT2316-7 occurred at high concentration of the nitrogen sources (monosodium glutamate or yeast extract). The capability to accumulate lipids with a fatty profile that can be tuned through cultivation temperature make the Antarctic thraustochytrid RT2316-13 a candidate for the production of lipids with different uses.© 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

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Investigating Meat Quality of Broiler Chickens Fed on Heat Processed Diets Containing Corn Distillers Dried Grains with Solubles.

The present study investigated the effects of feed form and distiller\'s dried grain with solubles (DDGS) on meat quality and fatty acids profile of broiler chickens. A total of 720 broilers (Ross 308; average BW [] 541±5.7 g) were randomly allotted to six treatments. Birds were fed three different feed forms (mash; SP, simple pellet; EP, expanded pellet) and DDGS (0 or 200 g kg) in a 3×2 factorial arrangement. The addition of DDGS and EP to the diet resulted in increased shear force of breast meat. Moreover, DDGS inclusion in the diet reduced the concentration of stearic and behenic in thigh meat. Pelleting (SP and EP) of the diets increased content in the thigh, whereas the linolenic content decreased. The breast mass was higher with EP and SP diets than with the mash diet. Feed processing led to increased pectoralis muscle and drum mass compared to mash-fed chickens. In conclusion, our results demonstrated that EP decreased thigh linolenic and meat shear force. In addition, DDGS supplementation in broiler hampers meat quality by increasing the shear force.

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Silymarin protects against renal injury through normalization of lipid metabolism and mitochondrial biogenesis in high fat-fed mice.

Obesity is associated with an increased risk of chronic kidney diseases and the conventional treatment with renin-angiotensin-aldosterone system (RAAS) inhibitors is not enough to prevent renal injury and prolong the progression of disease. Recently, silymarin has shown protective effects on renal tissue injury, but the underlying mechanisms remain elusive. The goal of this study was to investigate the potential capacity of silymarin to prevent renal injury during obesity induced by high fat diet (HFD) in mice. In vivo, male C57BL/6 mice received HFD (60% of total calories) for 12 weeks, randomized and treated orally with vehicle saline or silymarin (30mg/kg /d) for 4 weeks. In vitro, human proximal tubular epithelial cells (HK2) were exposed to 300μM (PA) for 36h followed by silymarin administration at different concentrations. The administration of silymarin significantly ameliorated HFD induced glucose metabolic disorders, oxidative stress and pathological alterations in the kidney. Silymarin significantly mitigated renal lipid accumulation, fatty β-oxidation and mitochondrial biogenesis in HFD mice and PA treated HK2 cells. Furthermore, silymarin partly restored mitochondrial membrane potential of HK2 cells after PA exposure. In conclusion, silymarin can improve oxidative stress and preserve mitochondrial dysfunction in the kidney, potentially via preventing accumulation of renal lipids and fatty β-oxidation.Copyright © 2017. Published by Elsevier Inc.

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Exercise Training Protects Cardiomyocytes from Deleterious Effects of Palmitate.

We investigated the effects of palmitate, a high saturated fat, on Ca, action potential and reactive oxygen species dynamics in cardiomyocytes from untrained and trained mice. Male mice were subjected to moderate intensity exercise training on a treadmill. Cardiomyocytes of untrained and trained mice were isolated, treated for 30\u2009min with palmitate and intracellular calcium transient and action potential duration were recorded. Additionally, we assessed reactive oxygen species generation. Treatment of cardiomyocytes from untrained mice with palmitate induced a significant decrease in Ca transient magnitude by 34%. Exercise training did not change cardiomyocyte Ca dynamics in the control group. However, trained cardiomyocytes were protected from deleterious effects of palmitate. Action potential duration was not altered by palmitate in either untrained or trained cardiomyocytes. Moreover, palmitate treatment increased reactive oxygen species generation in both untrained and trained cardiomyocytes. Nevertheless, the levels of reactive oxygen species in trained cardiomyocytes treated with palmitate were still 27% lower than those seen at basal conditions in untrained cardiomyocytes. Taken together, these findings indicate that exercise training protects cardiomyocytes from deleterious effects of palmitate possibly by inhibiting exacerbated ROS production.© Georg Thieme Verlag KG Stuttgart · New York.

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Nutritive Value of Desert Truffles Species of Genera Terfezia and Picoa (Ascomycetes) from Arid and Semiarid Regions of Eastern Turkey.

Terfezia and Picoa species contained 63-94 kcal energy, 76.93-83.81 g moisture, 0.78-1.52 g ash, 2.19-4.69 g protein, 2.14-9.48 g carbohydrate, 2.54-11.23 g dietary fiber, and 0.96-3.40 g fat per 100 g wet . We determined that Terfezia species contain more vitamin E and malondialdehyde than do Picoa species, but the amounts of vitamin A, vitamin C, and β-carotene may vary. Picoa species contain less and stearic acids but more oleic than do Terfezia species. Amino analyses revealed that glutamic and aspartic were the most abundant. We also noted that Picoa species have larger amounts of glucose and fructose than do Terfezia species. In addition, the elements these mushrooms contain can vary, but they are found at nutritious levels and are below toxic levels.

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Hemin Improves Insulin Sensitivity and Lipid Metabolism in Cultured Hepatocytes and Mice Fed a High-Fat Diet.

Hemin is a breakdown product of hemoglobin. It has been reported that the injection of hemin improves lipid metabolism and insulin sensitivity in various genetic models. However, the effect of hemin supplementation in food on lipid metabolism and insulin sensitivity is still unclear, and whether hemin directly affects cellular insulin sensitivity is yet to be elucidated. Here we show that hemin enhances insulin-induced phosphorylation of insulin receptors, Akt, Gsk3β, FoxO1 and cytoplasmic translocation of FoxO1 in cultured primary hepatocytes under insulin-resistant conditions. Furthermore, hemin diminishes the accumulation of triglyceride and increases in free fatty content in primary hepatocytes induced by palmitate. Oral administration of hemin decreases , energy intake, blood glucose and triglyceride levels, and improves insulin and glucose tolerance as well as hepatic insulin signaling and hepatic steatosis in male mice fed a high-fat diet. In addition, hemin treatment decreases the mRNA and protein levels of some hepatic genes involved in lipogenic regulation, fatty synthesis and storage, and increases the mRNA level and enzyme activity of CPT1 involved in fatty oxidation. These data demonstrate that hemin can improve lipid metabolism and insulin sensitivity in both cultured hepatocytes and mice fed a high-fat diet, and show the potential beneficial effects of hemin from food on lipid and glucose metabolism.

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Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk fat content (3.60 vs. 3.41%), milk fat yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% fat-corrected milk (47.6 vs. 46.5 kg/d), and energy-corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), (720 vs. 723 kg), or condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk fat yield (1.70 vs. 1.66 kg/d), and 3.5% fat-corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk fat yield, energy-corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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Stimulating effect of palmitate and insulin on cell migration and proliferation in PNT1A and PC3 prostate cells: Counteracting role of metformin.

A potential association between obesity and prostate cancer has been proposed. Metformin, an antidiabetes drug, has antiproliferative effects being proposed for cancer treatment. However, under intense proliferative stimulation conditions such as those found in obesity, its efficacy is still uncertain. Thus, we analyzed the effects of saturated fatty and/or insulin under high concentrations, with or without metformin, on the proliferation and migration of prostate cells.Human prostate epithelial cell lines non-tumor (PNT1A) and tumor (PC3) were treated with control media (DMEM, C), palmitate (100\u2009µM, HF), and/or insulin (50\u2009µU, HI) with or without metformin (100\u2009µM) for 24 or 48\u2009h.Both PNT1A and PC3 cells had greater proliferation when treated with HF, while HI treatment stimulated only PNT1A. Metformin inhibited cell proliferation caused by HF in both cell lines, but it did not block the proliferative action of HI in PNT1A cells. PNT1A increased cell migration after all treatments, while only HF influenced PC3; metformin inhibited the migration stimulated by all obese microenvironments. Both HF and HI treatments in PNT1A and HF treatment in PC3 augmented vimentin expression, resulting in a higher epithelial-mesenchymal transition (which, in turn, could influence cell migration). Metformin inhibited vimentin expression in both normal and tumor cells. Although HF treatment had increased AMPK activation, it also increased the levels of activated ERK1/2, which could be responsible for high cell proliferation in both cell lines. In contrast, HI decreased AMPK activation in both cell lines, whereas it increased ERK1/2 levels in PNT1A and decreased them in PC3 (reflecting greater cell proliferation only in non-tumor cells). Metformin maintained high activation of AMPK and decreased ERK1/2 levels after HF in both cell lines and only after HI in PNT1A, which was able to decrease the cell proliferation triggered by these treatments.Higher concentrations of palmitate on PC3 cells and palmitate and insulin on PNT1A cells stimulate cellular activities that could favor cancer progression. Metformin inhibited most of these stimuli, showing the efficacy of this drug for cancer adjuvant therapy in obese patients (a group at increased risk for the development of prostrate cancer).© 2018 Wiley Periodicals, Inc.

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Short communication: Effects of prill size of a -enriched fat supplement on the yield of milk and milk components, and nutrient digestibility of dairy cows.

The objective of our experiment was to evaluate the effects of prill size of a -enriched fatty supplement (PA; 85% C16:0) on feed intake, nutrient digestibility, and production responses of dairy cows. Twenty-four primiparous and multiparous Holstein cows were assigned based on parity and production level to replicated 4×4 Latin squares balanced for carryover effects with 21-d periods. Treatments were a control diet (no added PA), or 2.0% PA added as a small prill size (PA-SM; 284±12.4µm), a medium prill size (PA-MD; 325±14.7µm), or a large prill size (PA-LG; 600±17.4µm) supplement. Overall, PA treatments increased milk fat content (4.25 vs. 3.99%), milk fat yield (1.48 vs. 1.39kg/d), 3.5% fat-corrected milk (39.2 vs. 37.7kg/d), and improved feed efficiency (fat-corrected milk:dry matter intake; 1.51 vs. 1.42) compared with control. Compared with control, PA treatments did not affect dry matter intake, , condition score, or yields of milk, protein, and lactose. The PA treatments increased neutral detergent fiber digestibility (44.8 vs. 42.4%) and reduced the digestibility of 16-carbon fatty acids (72.3 vs. 79.1%) and total fatty acids (76.6 vs. 80.3%). Compared with control, PA treatments reduced the contents of de novo synthesized milk fatty acids (23.0 vs. 25.8g/100g of fatty acids) and preformed milk fatty acids (36.3 vs. 39.1g/100g of fatty acids), but did not affect their yields. In contrast, PA treatments increased the content (40.8 vs. 35.1g/100g of fatty acids) and yield (570 vs. 436g/d) of 16-carbon milk fatty acids compared with control. The PA prill size had no effect on dry matter intake, yield of milk and milk components, or feed efficiency. However, PA-LG tended to increase milk fat content compared with PA-SM (4.28 vs. 4.22%), and it increased 16-carbon fatty digestibility compared with PA-MD (74.2 vs. 71.0%) and PA-SM (74.2 vs. 71.7%). Additionally, PA-LG increased total fatty digestibility compared with PA-MD (78.1 vs. 75.6%) and PA-SM (78.1 vs. 76.0%). Results demonstrate that PA increased milk fat content and yield, and feed efficiency. Reducing prill size decreased fatty digestibility, but it had no effect on animal performance under the dietary conditions and prill sizes evaluated.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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Phthalate exposure and childhood overweight and obesity: Urinary metabolomic evidence.

Metabolomics may unravel global metabolic changes in response to environmental exposures and identify important biological pathways involved in the pathophysiology of childhood obesity. Phthalate has been considered an obesogen and contributing to overweight and obesity in children. The purpose of this study is to evaluate changes in urine metabolites in response to the environmental phthalate exposure among overweight or obese children, and to investigate the metabolic mechanisms involved in the obesogenic effect of phthalate on children at puberty.Within the national Puberty Timing and Health Effects in Chinese Children (PTHEC) study, 69 overweight/obese children and 80 normal children were selected into the current study according to their puberty timing and WGOC (The Working Group for obesity in China) references. Urinary concentrations of six phthalate monoesters (MMP, MEP, MnBP, MEHP, MEOHP and MEHHP) were measured using API 2000 electrospray triple quadrupole mass spectrometer (ESIMS/MS). Metabolomic profiling of spot urine was performed using gas chromatography-mass spectrometry. Differentially expressed urinary metabolites associated with phthalate monoesters exposure were examined using orthogonal partial least square-discriminant analysis and multiple linear regression models. In addition, the candidate metabolites were regressed to obesity indices with multiple linear regression models and logistic regression models in all subjects.Compared with normal children, higher levels of MnBP were detected in urinary samples of children with overweight and obesity. After adjusting for confounders including chronological age, gender, puberty onset, daily energy intake and physical activity and socio-economic level, positive association remained between urinary MnBP concentration and childhood overweight/obesity [OR\u202f=\u202f1.586, 95% CI:1.043,2.412]. We observed elevated MnBP concentration was significantly correlated with increased levels of monostearin, 1-monopalmitin, stearic , itaconic , glycerol 3-phosphate, 5-methoxytryptamine, kyotorphin, 1-methylhydantoin, d-alanyl-d-alanine, pyrrole-2-carboxylic , 3,4-Dihydroxyphenylglycol, and butyraldehyde. Meanwhile, increased MnBP concentration was also significantly correlated with decreased levels of lactate, glucose 6-phosphate, d-fructose 6-phosphate, , 4-acetamidobutyric , l-glutamic , n-acetyl-l-phenylalanine, iminodiacetic , hydroxyproline, pipecolinic , l-ornithine, n-acetyl-l-glutamic , guanosine, cytosin, and (s)-mandelic in the normal subjects. The observations indicated that MnBP exposure was related to global urine metabolic abnormalities characterized by disrupting arginine and proline metabolism and increasing oxidative stress and fatty reesterification. Among the metabolic markers related to MnBP exposure, 1-methylhydantoin, pyrrole-2-carboxylic and monostearin were found to be positively correlated with obesity indices, while hydroxyproline, l-ornithine, and lactate were negatively associated with overweight/obesity in children.Our results suggested that the disrupted arginine and proline metabolism associated with phthalate exposure might contribute to the development of overweight and obesity in school-age children, providing insights into the pathophysiological changes and molecular mechanisms involved in childhood obesity.Copyright © 2018 Elsevier Ltd. All rights reserved.

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Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes.

Type 2 diabetes mellitus is characterized by insulin resistance. However, the complete molecular mechanism remains unclear. In this study, zebrafish were fed a long-term high-fat diet to induce type 2 diabetes, which resulted in a higher , mass index, more lipid vacuoles in liver, increased insulin transcription level in liver, brain and muscle, and high fasting blood glucose in the high-fat diet zebrafish. Oppositely, the transcription levels of insulin substrate-2 and glucose transporter 2 were significantly decreased, indicating insulin signaling pathway and glucose transport impaired in the insulin-targeting tissues. Transcription of the autophagy-related genes, ATG3, ATG4B, ATG5, ATG7, ATG12, and FOXO3, were decreased but autophagy inhibitor gene m-TOR increased, and autophagy-flux was inhibited in liver of the high-fat diet zebrafish. Main of these changes were confirmed in -treated HepG2 cells. Further, in co-immunoprecipitation and subcellular co-localization experiments, the conjunction of preproinsulin with cargo-recognition protein p62 increased, but conjuncts of autophagosome with p62-cargo, lysosomes with p62-cargo, and autolysosomes decreased apparently. Interestingly, lysosomes, autolysosomes and conjuncts of p62-insulin localized at the periphery of -treated cells, the margination of lysosomes may mediate deactivation of proteases activity. These findings suggest that intracellular high-lipid may trigger defective autophagy, defective downstream signaling of insulin and accumulated intracellular preproinsulin, leading to dysregulation of cell homeostasis mechanism, which may be one of reasons involved in insulin-resistance in type 2 diabetes.

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Superhydrophobic Polypropylene Functionalized with Nanoparticles for Efficient Fast Static and Dynamic Separation of Spilled Oil from Water.

Frequent oil spills not only threaten the ecosystem, but they are also a waste of a valuable source of energy. There is an urgent need to develop materials that can readily remove spilled oil from water bodies and also have the capacity to collect it for energy applications. Herein, a superhydrophobic fiber of functionalized polypropylene is engineered with the help of interaction with incorporated copper oxide nanoparticles. The successful development of functionalized polypropylene is confirmed by Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, and energy-dispersive X-ray spectroscopy. The scanning electron microscopy images reveal that the surface roughness of the polypropylene is enhanced after functionalization. The optimized functionalized polypropylene displays an ultrahydrophobic surface with a water contact angle of 162.42°. The functionalized polyprolyene displays good absorption capacity. It has the capacity to take 30 to 40 times its own in oils and nonpolar organic solvents, which makes it useful for small spills. With a flux of 11 204 Lm h, functionalized polypropylene is as an ideal material for the dynamic separation of oil spills from water. It also has excellent selectivity towards oil, water rejection, and oil absorption capacity.© 2019 The Authors. Published by WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

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Pediatric Age Palm Oil Consumption.

Palm oil is widely used in the food industry for its chemical/physical properties, low cost and wide availability. Its widespread use has provoked an intense debate about whether it is a potential danger to human health. In a careful review of the scientific literature, we focused on nutritional characteristics and health effects of the use of palm oil with regards to children, seeking to determine whether there is evidence that justifies fears about the health effects of palm oil. Our review showed that palm oil represents a significant source of saturated fatty acids, to which scientific evidence attributes negative health effects when used in excess, especially with regards to cardiovascular diseases. However, to date, there is no evidence about the harmful effects of palm oil on the health of children. Nevertheless, palm oil has possible ill health effects linked to its composition of fatty acids: its consumption is not correlated to risk factors for cardiovascular diseases in young people with a normal and cholesterol level; the elderly and patients with dyslipidaemia or previous cardiovascular events or hypertension are at a greater risk. Therefore, the matter is not palm oil itself but the fatty--rich food group to which it belongs. The most important thing is to consume no more than 10% of saturated fatty acids, regardless of their origin and regardless of one\'s age. Correct information based on a careful analysis of the scientific evidence, rather than a focus on a singular presumed culprit substance, should encourage better lifestyles.

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Phytochemical Analysis and Potential Biological Activities of Essential Oil from Rice Leaf.

Although many investigations on phytochemicals in rice plant parts and root exudates have been conducted, information on the chemical profile of essential oil (EO) and potent biological activities has been limited. In this study, chemical compositions of rice leaf EO and in vitro biological activities were investigated. From 1.5 kg of fresh rice leaves, an amount of 20 mg EO was obtained by distillation and analyzed by gas chromatography-mass spectrometry (GC-MS), electrospray ionization (ESI), and atmospheric pressure chemical ionization (APCI) to reveal the presence of twelve volatile constituents, of which methyl ricinoleate (27.86%) was the principal compound, followed by (17.34%), and linolenic (11.16%), while 2-pentadecanone was the least (2.13%). Two phytoalexin momilactones A and B were first time identified in EO using ultra-performance liquid chromatography coupled with electrospray mass spectrometry (UPLC/ESI-MS) (9.80 and 4.93 ng/g fresh , respectively), which accounted for 7.35% and 3.70% of the EO, respectively. The assays of DPPH (IC = 73.1 µg/mL), ABTS (IC = 198.3 µg/mL), FRAP (IC = 700.8 µg/mL) and β-carotene oxidation (LPI = 79%) revealed that EO possessed an excellent antioxidant activity. The xanthine oxidase assay indicated that the anti-hyperuricemia potential was in a moderate level (IC = 526 µg/mL) as compared with the standard allopurinol. The EO exerted potent inhibition on growth of , and two noxious weeds , and , but in contrast, the growth of rice seedlings was promoted. Among the examined plants, the growth of the root was the most inhibited, proposing that constituents found in EO may have potential for the control of the problematic paddy weed . It was found that the EO of rice leaves contained rich phytochemicals, which were potent in antioxidants and gout treatment, as well as weed management. Findings of this study highlighted the potential value of rice leaves, which may provide extra benefits for rice farmers.

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The impact of obesity in the cardiac lipidome and its consequences in the cardiac damage observed in obese rats.

To explore the impact of obesity on the cardiac lipid profile in rats with diet-induced obesity, as well as to evaluate whether or not the specific changes in lipid species are associated with cardiac fibrosis.Male Wistar rats were fed either a high-fat diet (HFD, 35% fat) or standard diet (3.5% fat) for 6 weeks. Cardiac lipids were analyzed using by liquid chromatography-tandem mass spectrometry.HFD rats showed cardiac fibrosis and enhanced levels of cardiac superoxide anion (O), HOMA index, adiposity, and plasma leptin, as well as a reduction in those of cardiac glucose transporter (GLUT 4), compared with control animals. Cardiac lipid profile analysis showed a significant increase in triglycerides, especially those enriched with , stearic, and arachidonic . An increase in levels of diacylglycerol (DAG) was also observed. No changes in cardiac levels of diacyl phosphatidylcholine, or even a reduction in total levels of diacyl phosphatidylethanolamine, diacyl phosphatidylinositol, and sphingomyelins (SM) was observed in HFD, as compared with control animals. After adjustment for other variables (oxidative stress, HOMA, cardiac hypertrophy), total levels of DAG were independent predictors of cardiac fibrosis while the levels of total SM were independent predictors of the cardiac levels of GLUT 4.These data suggest that obesity has a significant impact on cardiac lipid composition, although it does not modulate the different species in a similar manner. Nonetheless, these changes are likely to participate in the cardiac damage in the context of obesity, since total DAG levels can facilitate the development of cardiac fibrosis, and SM levels predict GLUT4 levels.Copyright © 2017 Sociedad Española de Arteriosclerosis. Publicado por Elsevier España, S.L.U. All rights reserved.

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Diapause Induced by Temperature and Photoperiod Affects Fatty Compositions and Cold Tolerance of Phthorimaea Operculella (Lepidoptera: Gelechiidae).

To find out the potential condition for diapause induction in the potato tuber moth Phthorimaea operculella (Zeller), combination of constant temperatures (15, 20, and 30°C) and photoperiods (8, 9, 10, 11, 12, 14, and 16 h) were employed from egg to adult emergence. In addition, changes in the total phospholipid fatty content and cold tolerance of non-diapausing and diapausing pupae were examined. The critical daylength for diapause induction were 12.43 h at 20°C and lower temperatures that can induce in 50% of population. Moreover, the composition of total phospholipid fatty acids in the pupae revealed seven major fatty acids in both non-diapausing and diapausing pupae: oleic (26-32%), (21-29%), linoleic (18-21%), palmitoleic (4-10%), stearic (9%), linolenic (7-8%), and pentadecanoic acids (3-5%) with an increase in proportion of unsaturated fatty acids in diapause state. In contrast to increase of oleic (C18:1) from 26 to 32% in non-diapausing to diapausing pupae, a decrement trend from 29 to 21% in (C16:0) was observed at the same state. Additionally, supercooling point was observed to be significantly lower in diapausing (-22.6°C) than in non-diapausing pupae (-18.5°C) and the fresh of diapausing pupae was found to be significantly higher than non-diapausing ones. The significance of these findings would allow us a better understanding of interrelationship between diapause and cold tolerance.© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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Tuning culturing conditions towards the production of neutral lipids from lubricant-based wastewater in open mixed bacterial communities.

Production of bacterial lipid-based biofuels using inexpensive substrates, as wastes, is an emerging approach. In this work, a selective process using carbon feast-famine cycles was applied to obtain an indigenous microbial community of hydrocarbon-degrading and lipid-accumulating bacteria, using a real lubricant-based wastewater as carbon source. In the conditions applied, the enriched bacterial community, dominated by members of the genus Rhodococcus, Pseudomonas and Acinetobacter, was able to degrade almost all hydrocarbons present in the wastewater within 24\u202fh\' incubation and to accumulate, although in low levels, triacylglycerol (TAG) (<5% of cell dry (CDW)) and polyhydroxyalkanoates (PHA) (3.8%\u202f±\u202f1.1% of the CDW) as well as an unknown lipid (29%\u202f±\u202f6% of CDW), presumably a wax ester-like compound. The influence of culture conditions, namely carbon and nitrogen concentrations (and C/N ratio) and cultivation time, on the amount and profile of produced storage compounds was further assessed using a statistical approach based on a central composite circumscribed design and surface response methodology. The regression analysis of the experimental design revealed that only nitrogen concentration and C/N ratio are significant for neutral lipid biosynthesis (p\u202f<\u202f0.05). Maximum neutral lipid content, i.e. 33% (CDW basis), was achieved for the lowest carbon and nitrogen concentrations evaluated (10\u202fg COD L and 0.02\u202fg\u202fN L). PHA accounted for less than 5% of CDW. In these conditions, neutral lipid content was mainly composed by TAG, about 70% (w/w). TAG precursors, namely monoacylglycerols (MAG), diacylglycerols (DAG) and fatty acids (FA), accounted for 22% of total neutral lipids and WE for about 7%. Nevertheless, according to the applied response surface model, further improvement of neutral lipids content is still possible if even lower nitrogen concentrations are used. The fatty acids detected in TAG extracts ranged from myristic (C14:0) to linoleic (C18:2), being the most abundant (C16:0), stearic (C18:0) and oleic (C18:1). This study shows the feasibility of combining treatment of hydrocarbon contaminated wastewater, herein demonstrated for lubricant-based wastewater, with the production of bacterial neutral lipids using open mixed bacterial communities. This approach can decrease the costs associated to both processes and contribute to a more sustainable waste management and production of lipid-based biofuels.Copyright © 2018 Elsevier Ltd. All rights reserved.

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Mammary gland and milk fatty composition of two dairy goat breeds under feed-restriction.

Goat dairy products are an important source of animal protein in the tropics. During the dry season, pasture scarcity leads animals to lose up to 40% of their , a condition known as Seasonal Loss (SWL) that is one of the major constraints in ruminant production. Breeds with high tolerance to SWL are relevant to understand the physiological responses to pasture scarcity so they could be used in programs for animal breeding. In the Canary Islands there are two dairy goat breeds with different levels of tolerance to SWL: the Palmera, susceptible to SWL; and the Majorera, tolerant to SWL. Fat is one of the milk components most affected by environmental and physiological conditions. This study hypothesises that feed-restriction affects Majorera and Palmera breeds differently, leading to different fatty profiles in the mammary gland and milk. An interaction between breed and feed-restriction was observed in the mammary gland. Feed-restriction was associated with an increase in oleic and a decrease in percentage in the Palmera breed whereas no differences were observed in the Majorera breed. and oleic acids together constituted around 60% of the total fatty acids identified, which suggests that Palmera breed is more susceptible to SWL. In milk, feed-restriction affected both breeds similarly. Regarding the interaction of the breed with the treatment, we also observed similar responses in both breeds, but this influence affects only around 2% of the total fatty acids. In general, Majorera breed is more tolerant to feed-restriction.

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LC-MS/MS bioanalysis of plasma 1, 14-tetradecanedioic and 1, 16-hexadecanedioic as candidate biomarkers for organic anion-transporting polypeptide mediated drug-drug interactions.

A robust LC-MS/MS assay was developed to quantify endogenous 1, 14-tetradecanedioic (TDA) and 1, 16-hexadecanedioic (HDA) in human plasma as potential biomarkers for evaluating drug-drug interactions mediated by the hepatic drug transporters, organic anion-transporting polypeptides.This assay was validated using fit-for-purpose approach over standard curve range of 2.5-1000\xa0nM for TDA and HDA using analyte-free charcoal-stripped human plasma as the surrogate matrix. Chromatographic separation condition was successfully optimized to separate TDA from an interference peak while maintaining both analytes in neutral forms to minimize carryover issue.The described assay is currently applied to a clinical study for evaluating TDA/HDA as potential substitute biomarkers for drug-drug interaction studies.

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Depot-specific differences in fatty composition and distinct associations with lipogenic gene expression in abdominal adipose tissue of obese women.

Cardiometabolic diseases are primarily linked to enlarged visceral adipose tissue (VAT). However, some data suggest heterogeneity within the subcutaneous adipose tissue (SAT) depot with potential metabolic differences between the superficial SAT (sSAT) and deep SAT (dSAT) compartments. We aimed to investigate the heterogeneity of these three depots with regard to fatty (FA) composition and gene expression. Adipose tissue biopsies were collected from 75 obese women undergoing laparoscopic gastric bypass surgery. FA composition and gene expression were determined with gas chromatography and quantitative real-time-PCR, respectively. Stearoyl CoA desaturase-1 (SCD-1) activity was estimated by product-to-precursor FA ratios. All polyunsaturated FAs (PUFA) with 20 carbons were consistently lower in VAT than either SAT depots, whereas essential PUFA (linoleic , 18:2n-6 and α-linolenic , 18:3n-3) were similar between all three depots. Lauric and were higher and lower in VAT, respectively. The SCD-1 product palmitoleic as well as estimated SCD-1 activity was higher in VAT than SAT. Overall, there was a distinct association pattern between lipid metabolizing genes and individual FAs in VAT. In conclusion, SAT and VAT are two distinct depots with regard to FA composition and expression of key lipogenic genes. However, the small differences between sSAT and dSAT suggest that FA metabolism of SAT is rather homogenous.

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Qualitative and quantitative comparison of cyclic phosphatidic and its related lipid species in rat serum using hydrophilic interaction liquid chromatography with tandem-mass spectrometry.

Cyclic phosphatidic (cPA) is a simple lipid containing a fatty attached at the sn-1 position and a cyclic phosphate ring structure at the sn-2 and sn-3 positions of the glycerol backbone. The pharmacological effects of cPA have been demonstrated in several diseases such as cancer and neuropathic pain; however, the composition of the molecular species of cPA in relative to other lipid species in biological samples is still unclear. Recently, hydrophilic interaction liquid chromatography (HILIC) has demonstrated the ability to perform lipidomic analyses of biological samples. In the present study, we developed the quantitative measurement of cPA and its related lipid species, such as lysophosphatidic (LPA) and lysophosphatidylcholine (LPC), in rat serum using HILIC equipped with tandem-mass spectrometry (MS/MS). The lipid analysis using HILIC-MS/MS system demonstrated high linearity and reproducibility. The modified Bligh and Dyer method using citric was showed high efficiency on the extraction of cPA and LPA without contamination of artificial products. In rat serum, cPA and LPC contained more saturated fatty acids such as and stearic than unsaturated fatty acids, whereas LPA and phosphatidylcholine more contained unsaturated fatty acids than saturated fatty acids. The analytical methods for measuring cPA and its related lipid species in the present study will aid the analysis of their metabolism.Copyright © 2018 Elsevier B.V. All rights reserved.

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Acanthoic modulates lipogenesis in nonalcoholic fatty liver disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic fatty liver disease (NAFLD) associated with lipid accumulation by activating Farnesoid X receptor (FXR) and liver X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high-fat (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on lipid accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in , liver index, and liver lipid droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated lipid accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating lipid accumulation and fatty synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

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pH effects on the lipid and fatty acids accumulation in Chlamydomonas reinhardtii.

pH variations influence the delivery of essential nutrients and CO solubility, which impact algae metabolism. In this study the microalgal growth and chlorophyll, lipid, and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. Lipid content increased 3.0 (14.81% of dry cell , dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for ; 17.3, 14.7, and 25.7% for oleic ; and 9.8, 12.1, and 4.6% for linoleic ; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of lipid biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and lipid accumulation.© 2019 American Institute of Chemical Engineers.

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Effects of a co-micronized composite containing palmitoylethanolamide and polydatin in an experimental model of benign prostatic hyperplasia.

Palmitoylethanolamide (PEA), a fatty amide-signaling molecule has well-known anti-inflammatory and neuroprotective effects. Nevertheless, PEA does not possess the ability to prevent free radicals formation. Polydatin (PLD), a biological precursor of resveratrol, has antioxidant activity. A combination of PEA and PLD could, conceivably, have beneficial effects on oxidative stress produced by inflammatory processes. In the present study we investigated the effects of a co-micronized composite containing PEA and PLD (m(PEA/PLD)) in a model of testosterone-induced benign hyperplasia (BPH). BPH was provoked in rats by daily administration of testosterone propionate (3mg/kg) for 14days. This protocol leads to alterations in prostate morphology and increased levels of prostaglandin E2 and dihydrotestosterone as well as of 5α-reductase 1 and 5α-reductase 2 expression. Moreover, testosterone induced marked inflammation in terms of an increase in nuclear translocation of nuclear factor-κB p65 and consequently in IκB-α degradation as well as disregulation of inducible nitric oxide synthase, cyclooxygenase-2 and manganese superoxide dismutase expression and in the apoptosis pathway. Our results show, for the first time, that m(PEA/PLD) is capable of decreasing prostate and dihydrotestosterone production in BPH-induced rats. These effects were most likely correlated to the anti-inflammatory and apoptotic effects of m(PEA/PLD). Accordingly, these results support the view that m(PEA/PLD) should be further studied as a potent candidate for the management of BPH.Copyright © 2017 Elsevier Inc. All rights reserved.

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Lipid profile changes in erythrocyte membranes of women with diagnosed GDM.

Gestational diabetes mellitus (GDM) is a glucose intolerance that begins or is first recognized during pregnancy. It is currently a growing health problem worldwide affecting from 1% to 14% of all pregnant women depending on racial and ethnic group as well as the diagnostic and screening criteria. Our preliminary study aimed at investigating the erythrocyte membrane fatty profiles of pregnant women, in particular with diagnosed with gestational diabetes mellitus (GDM), and with normal glucose tolerant (NGT) pregnant women as a control group. The study group comprised 43 pregnant women, 32 of whom were diagnosed with GDM according to the WHO criteria, and 11 with normal glucose tolerance. The erythrocyte membrane phospholipids were obtained according to the Folch extraction procedure. Fatty acids (FA) were analyzed by gas chromatography (GC) as the corresponding fatty methyl esters (FAME). A cluster of 14 fatty acids identified contained >98% of the recognized peaks in the GC analysis. The analysis of fatty acids from erythrocytes revealed important differences between GDM and NGT women in the third trimester, and the results were correlated with biochemical data. Among the 14 measured FA representing the membrane lipidomic profile, the levels of three saturated FA (myristic, , stearic acids) tended to decrease in GDM patients, with the percentage content of stearic significantly changed. The relative content of monounsaturated fatty acids (MUFA) tended to increase, in particular the oleic and vaccenic contents were significantly increased in erythrocyte membranes of the GDM group in comparison with the NGT group. The GDM group demonstrated higher sapienic levels (+29%) but this change was not statistically significant. This study revealed association between an impaired cis-vaccenic concentration in erythrocytes membrane and GDM development. No significant changes of polyunsaturated fatty acids (PUFA) were observed in GDM and NGT erythrocytes. We postulate, basing on the differences between the GDM and NGT lipidomic profiles, that stearic and cis-vaccenic acids can be considered as dual biomarkers of specific SFA-MUFA conversion pathway, involving the coupling of delta-9 desaturase and elongase enzymes. Our results indicate that the SFA-MUFA families may be involved in the pathophysiology of metabolic diseases such as GDM, but the further studies are needed to confirm our hypothesis. In conclusion, the erythrocyte membranes of GDM women undergo remodeling resulting in abnormal fatty profiles, which are reflection of the long-term status of organism and can have great impact on both the mother and her offspring.

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Effects of fundamental nutrient stresses on the lipid accumulation profiles in two diatom species Thalassiosira weissflogii and Chaetoceros muelleri.

Microalgae are considered as attractive feedstocks for biofuel production nowadays because of their high lipid contents and easy cultivation. In the present study, two diatoms, Thalassiosira weissflogii and Chaetoceros muelleri, were cultured under various nutrient-limitation conditions to explore their comprehensive lipid accumulation profiles for further commercialization. In T. weissflogii, the highest neutral lipid accumulation and highest lipid productivity (14.28\xa0mg\xa0L\xa0day) were both recorded under P-limitation. In C. muelleri, the highest lipid content (35.03% of dry cell ), highest neutral lipid accumulation, and highest lipid productivity (29.07\xa0mg\xa0L\xa0day) were all recorded under N-limitation. Besides, the predominant fatty acids of T. weissflogii and C. muelleri were myristic (C14:0), (C16:0), and palmitoleic (C16:1), with the amounts of 58.4-74.4 and 74.1-87.7% of the total fatty acids, respectively. Moreover, nutrient limitations led to a lower proportion of polyunsaturated fatty acids (PUFA) than that of saturated fatty (SFA) and monounsaturated fatty (MUFA) in both species. The ratios of (SFA\u2009+\u2009MUFA) to PUFA were from 1.65 to 3.01 in T. weissflogii, and up to 3.61 to 8.59 in C. muelleri. Our results suggested the feasibility of C. muelleri as biodiesel feedstock due to its more suitable fatty composition and higher lipid productivity compared to T. weissflogii.

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Obesity-related cellular stressors regulate gonadotropin releasing hormone gene expression via c-Fos/AP-1.

Obesity is a risk factor for infertility, but mechanisms underlying this risk are unclear. Fertility is regulated by hypothalamic gonadotropin-releasing hormone, encoded by the Gnrh1 gene. Because obesity promotes endoplasmic reticulum (ER) stress, we sought to determine how tunicamycin-induced ER stress affected Gnrh1 gene expression in the mouse hypothalamic cell line GT1-7. Tunicamycin repressed expression of Gnrh1 in a PKC- and JNK-dependent manner, while upregulating expression of a known Gnrh1 repressor, Fos. Obesity is associated with increased circulating free fatty acids, and exposure to palmitate promoted ER stress and inflammation. Fos expression increased with palmitate dose, but Gnrh1 expression was upregulated with low-dose palmitate and repressed with high-dose palmitate. Using a small molecule inhibitor, we determined that AP-1 was required for Gnrh1 repression by high-dose palmitate or tunicamycin-induced ER stress. These findings suggest that hypogonadism driven by decreased hypothalamic GnRH may be a component of obesity-related infertility.Copyright © 2018 Elsevier B.V. All rights reserved.

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Enhanced production of carotenoids using a Thraustochytrid microalgal strain containing high levels of docosahexaenoic -rich oil.

Results to date suggest that microalgal Thraustochytrids family strains can be used to produce high-functional omega-3 rich oil (~\u200930-70% of dry cell ) and carotenoid-based antioxidant pigments simultaneously with value-added bioactive potential. In the present study, we describe the isolation and characterization of a new Thraustochytrid Schizochytrium sp. from the west coastal area of Korea. This newly isolated Thraustochytrid, identified as Schizochytrium sp. through 18S rRNA analysis and named SH104, simultaneously produces high levels of DHA and carotenoid-based antioxidant pigments. An improved Schizochytrium mutant, named SHG104, was obtained from the original host strain by γ-irradiation-induced mutagenesis. Under combined temperature-shift cultivation conditions employing white-light LEDs (light-emitting diodes), Schizochytrium sp. SHG104 yielded 10.8\xa0g L of biomass comprising 45.8% total lipids (32.1% DHA) and 4.6\xa0mg L of astaxanthin. In addition to DHA, the main fatty acids produced by Schizochytrium sp. SHG104 were and a trace of other long-chain fatty acids. The carotenoid profile of SH104 and SHG104 was β-carotene, astaxanthin, canthaxanthin, pheonicoxanthin and echinenone, which analyzed by HPLC and LC/APCI-MS. Furthermore, genomic analysis of Schizochytrium and Aurantiochytrium microalgae confirmed that the presence of carotenogenesis pathway enzymes and genes including geranylgeranyl diphosphate, phytoene synthase, lycopene cyclase, and cytochrome P450 hydroxylase that necessary for the production of antioxidants via a complete biosynthetic KEGG synthesis pathway. This newly isolated Schizochytrium microalga potentially have wide application as a source of antioxidants for astaxanthin-containing pigments, commercial omega-3 lipids and feed additives, such as nutritional supplements for aquaculture.

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Demonstration of bioprocess factors optimization for enhanced mono-rhamnolipid production by a marine Pseudomonas guguanensis.

We identified that Pseudomonas guguanensis produced macromolecular mono-rhamnolipid (1264.52\u202fDa) upon sensing n-hexadecane/diesel/kerosene from its surroundings. Permutation experiments were done to improve the laboratory-scale mono-rhamnolipid production (ie, a three-fold increase) using RSM validation. Consequently, maximal mono-rhamnolipids production [40-50\u202fmg/L] and emulsification abilities [65-70%] were encountered on day 8 using vegetable oil, peptone\u202f+\u202fyeast extract. EI values for the rhamnolipids were found to be 78±1.75% at 12.5\u202fmg/\u202fmL. Production and secretion of rhamnolipids were accompanied by aggregation of cells at day 6 as pictured in SEM. Pure monorhamnolipids of P. guguanensis was found to lower the surface tension of water to 32.98±0.3\u202fmN/m than the crude and CFSs of P. aeruginosa indicating efficient activity. Utilization and subsequent removal of hexadecane was 77.2% and the breakdown products were fatty acids [decanoic, hexadecanoic, octadecanoic acids and methyl stearates] as signified in Head-space GC-MS. The breakdown products of hexadecane are also present in the synthesized rhamnolipids suggesting their biosynthetic role. Rapid degradation of hexadecane, diesel and kerosene by this emulsifier combined with non-pathogenic trait of P. guguanensis identifies this organism as a viable option to remove n-alkanes from aquatic environments.Copyright © 2017 Elsevier B.V. All rights reserved.

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In vivo kinetic study of materno-fetal fatty transfer in obese and normal pregnant women.

Placental structure and function can be modified as a result of maternal obesity affecting materno-fetal fatty acids (FA) transport. We report for the first time, in humans and in vivo, the kinetics of placental FA transfer in normo- and in normolipemic obese pregnant women using stable isotopes. The administration of different tracer FA with similar behaviour to the mother at different time points allows the collection of kinetic information on materno-fetal transfer of FA despite only one sample of placenta and cord can be collected per subject. Computational modelling showed a good fit to the data when considering all maternal plasma lipid classes but not when based only on non-esterified FA. The novel approach using multiple tracer FA administration combined with computational modelling shows a consistent time course of placental tracer FA and predicted total FA accumulation.We analyse for the first time the in vivo materno-fetal kinetic transfer of fatty acids (FA) labelled with stable isotopes in control and obese (OB) pregnant women. Labelled FA with a similar metabolism (stearic : C-SA; : C-PA; oleic : C-OA) were orally administered at -4\xa0h, -8\xa0h and -12\xa0h, respectively prior to elective caesarean section to 10 pregnant women with a mass index >30 (OB) and 10 with a mass index in the range 20-25 (NW). Placenta, venous and arterial cord blood were collected obtaining a wide range of FA enrichments. A combined experimental and computational modelling analysis was applied. FA fractional synthesis rate (FSR) in placenta was 11-12%\xa0h . No differences were observed between NW and normo-lipidemic OB. It was not possible to estimate FA FSR in cord blood with this oral bolus dose approach. Computational modelling demonstrated a good fit to the data when all maternal plasma lipid classes were included but not with modelling based only on the non-esterified FA fraction. The estimated materno-fetal C-FA transfer was ∼1%. In conclusion, our approach using multiple C-FA tracers allowed us to estimated FSR in placental/maternal plasma but not in fetal/maternal compartments. Computational modelling showed a consistent time course of placental C-FA transfer and predicted total fetal FA accumulation during the experiment. We conclude that, in addition to non-esterified FA fraction in the maternal circulation, maternal plasma very low-density lipoprotein and other lipoproteins are important contributors to placental FA transfer to the fetus.© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.

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7-Hydroxymatairesinol improves , fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg 7-HMR (HMRLignan™) or 10 mg/kg TEP by oral administration. 7-HMR and TEP limited the increase in (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the liver was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and obesity in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

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Alternation of plasma fatty acids composition and desaturase activities in children with liver steatosis.

The aim of this study was to investigate changes in plasma fatty acids proportions and estimated desaturase activities for variable grading of liver steatosis in children.In total, 111 schoolchildren (aged 8-18 years) were included in the analysis from March 2015 to August 2016. Anthropometric evaluation, liver ultrasound examination and scoring for nonalcoholic fatty liver disease (NAFLD score = 0-6), and biochemical and plasma fatty acids analysis were performed. We compared the composition ratio of fatty acids between children with high-grade liver steatosis (NAFLD score = 4-6), low-grade liver steatosis (NAFLD score = 1-3), and healthy controls (NAFLD score = 0). In addition, correlation coefficients (r) between NAFLD score, metabolic variables, and estimated activity of desaturase indices (stearoyl-coenzyme A desaturase-1 (SCD1), delta-5 and delta-6 desaturase) were calculated.Compared with healthy controls, children with liver steatosis showed a higher proportion of monounsaturated fatty acids (21.16 ± 2.81% vs. 19.68 ± 2.71%, p = 0.024). In addition, children with high- grade liver steatosis exhibited higher proportions of (C16:0), palmitoleic (C16:1n-7), dihomo-γ-linolenic (C20:3n-6), adrenic (C22:4n-6), and docosapentaenoic (C22:5n-6); and lower proportions of eicosapentaenoic (C20:5n-3) (P< 0.05). In all subjects, the NAFLD score was positively correlated with mass index (BMI) (kg/m2) (r = 0.696), homeostasis model of assessment ratio-index (HOMA-IR) (r = 0.510), SCD1(16) (r = 0.273), and the delta-6 index (r = 0.494); and inversely associated with the delta-5 index (r = -0.443).Our current data suggested that children with liver steatosis was highly associated with obesity, and insulin resistance. In addition, increased endogenous lipogenesis through altered desaturase activity may contribute to the progression of liver steatosis in children.

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Feeding of palm oil fatty acids or rapeseed oil throughout lactation: Effects on energy status, composition, and milk production in Norwegian dairy goats.

The objective of this experiment was to examine how supplements of rapeseed oil or palm oil fatty acids would affect milk production and composition, lipid stores, and energy balance in 30 multiparous goats of Norwegian dairy goat breed. The experiment lasted 230 d, with 1 to 120 d in milk (DIM) for indoor feeding (P1), 120 to 200 DIM for mountain grazing (P2), and 200 to 230 DIM for indoor feeding (P3). Grass silage was fed according to appetite during indoor feeding periods. After an adjustment period (1-60 DIM) when the control diet was given to the goats, the animals were subdivided into 3 groups of 10 goats. Treatments (60-230 DIM) were (1) basal concentrate (control; no added fat); (2) control concentrate with 8% (added on air-dry basis) hydrogenated palm oil enriched with (POFA); and (3) control concentrate with 8% (added on air-dry basis) rapeseed oil (RSO). Individual energy balances based on energy intake and milk production were estimated on 10, 30, 60, 90, 120, 200, and 230 DIM. At the same times, (BW), condition score (BCS), mass index, and tissue stores using computed tomography were monitored. Silage intake was depressed by POFA throughout the experimental period. Reduced BW and mass index were observed in the POFA and RSO groups, whereas no effect on BCS or composition was observed throughout lactation. Generally, a minor decrease in BW was observed from 10 to 120 DIM (only 0.6 kg on average) and the total amount of lipid was reduced by 4.4 kg. During the mountain grazing period, a further reduction in lipid stores (2.7 kg) was observed, and BW was reduced by 3.9 kg in the same period. The goats mobilized, on average, 72% of their fat reserves during the first 200 DIM. In this period, dietary fat supplementation did not reduce the mobilization of adipose tissue but resulted in greater milk fat yield (2 kg more, on average, compared with the control group). Milk yield was not affected by POFA or RSO supplementation. Milk fat content was higher in the POFA group than in the control and RSO groups. Milk protein and lactose contents were not affected by lipid supplements. In late lactation, a rapid accumulation of fat deposits followed the intense mobilization during the grazing period. Dietary lipid supplements had no effect on milk fat yield at this stage. Milk production depends heavily on the ability to mobilize lipid stores, and neither POFA nor RSO supplements at rates used in our study affected this mobilization.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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Dihydrosterculic from cottonseed oil suppresses desaturase activity and improves liver metabolomic profiles of high-fat-fed mice.

Polyunsaturated fatty (PUFA)-rich diets are thought to provide beneficial effects toward metabolic health in part through their bioactive properties. We hypothesized that increasing PUFA intake in mice would increase peroxisome proliferator activated receptor delta (PPARδ) expression and activity, and we sought to examine the effect of different PUFA-enriched oils on muscle PPARδ expression. One of the oils we tested was cottonseed oil (CSO) which is primarily linoleic (53%) and (24%). Mice fed a CSO-enriched diet (50% energy from fat) displayed no change in muscle PPARδ expression; however, in the liver, it was consistently elevated along with its transcriptional coactivator Pgc-1. Male mice were fed chow or CSO-, saturated fat (SFA)-, or linoleic (18:2)-enriched diets that were matched for macronutrient content for 4 weeks. There were no differences in food intake, , fasting glucose, glucose tolerance, or energy expenditure between chow- and CSO-fed mice, whereas SFA-fed mice had increased fat mass and 18:2-fed mice were less glucose tolerant. Metabolomic analyses revealed that the livers of CSO-fed mice closely matched those of chow-fed but significantly differed from SFA- and 18:2-enriched groups. Fatty composition of the diets and livers revealed an impairment in desaturase activity and the presence of dihydrosterculic (DHSA) in the CSO-fed mice. The effect of DHSA on PPARδ and stearoyl-CoA desaturase-1 expression mimicked that of the CSO-fed mice. Taken together, these data suggest that DHSA from CSO may be an effective means to increase PPARδ expression with concomitant suppression of liver stearoyl-CoA desaturase-1 activity.Copyright © 2017 Elsevier Inc. All rights reserved.

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Effect of loss on circulating fatty profiles in overweight subjects with high visceral fat area: a 12-week randomized controlled trial.

Significant associations between visceral fat and alterations in plasma fatty acids have been identified in overweight individuals. However, there are scant data regarding the relationships of the visceral fat area (VFA) with the plasma fatty profiles and desaturase activities following loss. We investigated the effect of loss with mild calorie restriction on the circulating fatty profiles and desaturase activities in nondiabetic overweight subjects with high VFA.Eighty overweight subjects with high VFA (L4 VFA ≥100\xa0cm) were randomized into the 12-week mild-calorie-restriction (300\xa0kcal/day) or control groups.Comparison of the percent of changes between groups revealed that the -loss group had greater reductions in . The VFA decreased by 17.7\xa0cm from baseline in the -loss group\xa0(P < 0.001). At follow-up, the -loss group showed greater reductions in serum triglycerides, insulin, and HOMA-IR than the control group. Significantly greater reductions in total saturated fatty acids, , stearic , total monounsaturated fatty acids, palmitoleic , oleic , eicosadienoic , and dihomo-γ-linolenic levels were detected in the -loss group compared with the control group after adjusting for baseline values. Following loss, C16 Δ9-desaturase activity was significantly decreased and Δ5-desaturase activity was significantly increased, and the changes were greater in the -loss group than in the control group.The results suggest that mild loss improves abdominal obesity, overall fatty profiles, and desaturase activities; therefore, mild calorie restriction has potential health benefits related to obesity-related diseases in overweight subjects with high VFA.. Retrospectively registered 11 December 2016.

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Role of the mTOR‑FOXO1 pathway in obesity‑associated renal tubulointerstitial inflammation.

Since obesity is largely responsible for the growing incidence of renal tubulointerstitial inflammation, exploration into the mechanisms of obesity‑associated tubulointerstitial inflammation is essential. Studies have demonstrated that mammalian target of rapamycin\xa0(mTOR) is a crucial molecule in the pathogenesis of renal inflammation, including regulating the expression of inflammatory factors. The purpose of the present study was to further elucidate the role of mTOR in obesity‑associated tubulointerstitial inflammation. In the clinical study, obese and healthy subjects were recruited for physical examination, as well as the collection of blood and urine samples. Further study was performed on a high fat diet\xa0(HFD)‑induced obese rat model and a cultured human renal tubular epithelial cell line\xa0(HK‑2). The clinical study demonstrated that the participants with obesity had increased serum lipids, creatinine\xa0(Cr), urinary albumin to creatinine ratio\xa0(UACR) and urinary neutrophil gelatinase‑associated lipocalin\xa0(u‑NGAL). Moreover, the level of urinary monocyte chemoattractant protein‑1\xa0(u‑MCP‑1) was increased in the participants with obesity, and it was positively correlated with free fatty \xa0(FFA), UACR and u‑NGAL. In the in\xa0vivo study, the results indicated that the levels of serum lipids, Cr and blood urea nitrogen\xa0(BUN), as well as 24\xa0h urine protein and u‑NGAL, were significantly increased in the HFD‑fed obese rats. In addition, the infiltration of CD68+ cells into the renal interstitial area and the release of interleukin‑1β\xa0(IL‑1β) was observed in the kidneys of obese rats. Meanwhile, the supernatant from HK‑2 cells treated with stimulated THP‑1 monocyte migration. The upregulation of MCP‑1, phosphorylated forkhead boxO1 (p‑FOXO1), and phosphorylated mTOR (p‑mTOR) was observed in\xa0vivo and in\xa0vitro. However, inhibition of mTOR was able to alleviate the above effects. Overall, these results demonstrated that activated mTOR induced FOXO1 phosphorylation, which mediates renal MCP‑1 release, causes tubulointerstitial inflammation and ultimately leads to pathological renal changes and dysfunction. However, inhibition of mTOR may play a renoprotective role during the progression of obesity‑associated tubulointerstitial inflammation.

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Human milk enriched with human milk lyophilisate for feeding very low birth preterm infants: A preclinical experimental study focusing on fatty profile.

Human milk, with essential nutrients and long chain polyunsaturated fatty acids (LC-PUFAs) such as the omega 3 and 6 fatty acids is important for development of the central nervous system and the retina in very low birth infants (<1,500 g). However, breast milk may not be sufficient to meet these needs. The possibility of supplementing breast milk with a lyophilisate of human milk was explored in this study. The objectives of this study were to determine the total lipid content and the lipid profile of the Human Milk on Baseline (HMB) and that of the Concentrates with the Human Milk + lyophilisate (with lyophilisate of milk in the immediate period (HMCI), at 3 months (HMC3m), and at 6 months (HMC6m) of storage).Fifty donors from the Human Milk Bank of Children\'s Hospital provided consent, and donated milk samples. Macronutrient (including total lipids) quantification was performed using the MIRIS® Human Milk Analyzer, and the fatty profile was determined by gas chromatography (CG-FID, SHIMADZU®).There was a higher lipid concentration in HMCI relative to HMB. The concentrations of the main fatty acids (% of total) were as follows: (C16:0) HMB, 22.30%; HMCI, 21.46%; HMC3m, 21.54%; and HMC6m, 21.95% (p<0.01); oleic (C18:1n-9) HMB, 30.41%; HMCI, 30.47%; HMC3m, 30.55%; and HMC6m, 29.79% (p = 0.46); linoleic (C18:2n-6) HMB, 19.62%; HMCI, 19.88%; HMC3m, 19.49%; and HMC6m, 19.45% (p = 0.58); arachidonic (C20:4n-6) HMB, 0.35%; HMCI, 0.16%; HMC3m, 0.13%; and HMC6m, 0.15% (p<0.01); α-linolenic (C18:3n-3) HMB,1.32%; HMCI, 1.37%; HMC3m, 1.34%; and 1.34% HMC6m (p = 0.14); docosahexaenoic (C22:6n-3) HMB, 0.10%; HMCI, 0.06%; HMC3m, 0.05%; and HMC6m, 0.06% (p<0.01). There were no significant changes in the lipid profile when stored. There was no evidence of peroxidation during storage.Freeze-dried human milk fortified with a human milk concentrate brings potential benefits to newborns, mainly by preserving the essential nutrients present only in breast milk; however, further clinical studies are required to evaluate the safety and efficacy of the concentrate as a standard nutritional food option for very low birth infants.

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Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high-fat diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg ) for four weeks, the liver, liver/, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high-fat diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high-fat diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

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Fatty composition from the marine red algae Pterocladiella capillacea (S. G. Gmelin) Santelices & Hommersand 1997 and Osmundaria obtusiloba (C. Agardh) R. E. Norris 1991 and its antioxidant activity.

This study evaluated the chemical composition and antioxidant activity of fatty acids from the marine red algae Pterocladiella capillacea (S. G. Gmelin) Santelices & Hommersand 1997 and Osmundaria obtusiloba (C. Agardh) R. E. Norris 1991. The gas chromatography mass spectrometry (GC-MS) identified nine fatty acids in the two species. The major fatty acids of P. capillacea and O. obtusiloba were , oleic , arachidonic and eicosapentaenoic . The DPPH radical scavenging capacity of fatty acids was moderate ranging from 25.90% to 29.97%. Fatty acids from P. capillacea (31.18%) had a moderate ferrous ions chelating activity (FIC), while in O. obtusiloba (17.17%), was weak. The ferric reducing antioxidant power (FRAP) of fatty acids from P. capillacea and O. obtusiloba was low. As for β-carotene bleaching (BCB), P. capillacea and O. obtusiloba showed a good activity. This is the first report of the antioxidant activities of fatty acids from the marine red algae P. capillacea and O. obtusiloba.

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Overexpression of apolipoprotein A-I alleviates endoplasmic reticulum stress in hepatocytes.

Abnormal lipid metabolism may contribute to an increase in endoplasmic reticulum (ER) stress, resulting in the pathogenesis of non-alcoholic steatohepatitis. Apolipoprotein A-I (apoA-I) accepts cellular free cholesterol and phospholipids transported by ATP-binding cassette transporter A1 to generate nascent high density lipoprotein particles. Previous studies have revealed that the overexpression of apoA-I alleviated hepatic lipid levels by modifying lipid transport. Here, we examined the effects of apoA-I overexpression on ER stress and genes involved in lipogenesis in both HepG2 cells and mouse hepatocytes.Human apoA-I was overexpressed in HepG2 hepatocytes, which were then treated with 2\xa0μg/mL tunicamycin or 500\xa0μM . Eight-week-old male apoA-I transgenic or C57BL/6 wild-type mice were intraperitoneally injected with 1\xa0mg/kg tunicamycin or with saline. At 48\u2009h after injecting, blood and liver samples were collected.The overexpression of apoA-I in the models above resulted in decreased protein levels of ER stress makers and lipogenic gene products, including sterol regulatory element binding protein 1, fatty synthase, and acetyl-CoA carboxylase 1. In addition, the cellular levels of triglycerides and free cholesterol also decreased. Some of gene products which are related to ER stress-associated apoptosis were also affected by apoA-I overexpression. These results suggested that apoA-I overexpression could reduce steatosis by decreasing lipid levels and by suppressing ER stress and lipogenesis in hepatocytes.ApoA-I expression could significantly reduce hepatic ER stress and lipogenesis in hepatocytes.

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Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced obesity.

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti-obesity drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased and liver , insulin, leptin, triglyceride, cholesterol and free fatty plasma levels in a mouse model of diet-induced obesity. Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in energy expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in obesity and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

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Ceramide stearic to ratio predicts incident diabetes.

Ceramide lipids have a role in the development of insulin resistance, diabetes and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident diabetes.A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the metabolic syndrome (Prevent Metabolic Syndrome [PrevMetSyn], n\u2009=\u2009371). Diabetes risk score models were developed to estimate the 10\xa0year risk of incident diabetes.Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident diabetes (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and diabetes risk scores significantly decreased in individuals who had 5% or more loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident diabetes, and may be modulated by lifestyle intervention.

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Acute and Repeated Treatment with 5-PAHSA or 9-PAHSA Isomers Does Not Improve Glucose Control in Mice.

Fatty esters of hydroxylated fatty acids (FAHFAs) were discovered as a novel class of endogenous mammalian lipids whose profound effects on metabolism have been shown. In the current study, in\xa0vitro and in\xa0vivo the metabolic effects of two of these FAHFAs, namely -5- (or -9) -hydroxy-stearic (5- or 9-PAHSA, respectively) were profiled. In DIO mice fed with differentially composed low- or high-fat diets, acute and subchronic treatment with 5-PAHSA and 9-PAHSA alone, or in combination, did not significantly improve the deranged metabolic status. Neither racemic 5- or 9-PAHSA, nor the enantiomers were able to: (1) increase basal or insulin-stimulated glucose uptake in\xa0vitro, (2) stimulate GLP-1 release from GLUTag cells, or (3) induce GSIS in rat, mouse, or human islets or in a human pancreatic β cell line. Therefore, our data do not support the\xa0further development of PAHSAs or their derivatives for the control of insulin resistance and hyperglycemia.Copyright © 2018 Elsevier Inc. All rights reserved.

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[Research on influence of environment factors to yield and quality traits of Perilla frutescen].

The research is aimed to study of the influence of environmental factors on the yield and quality traits, and find out the regularity of the growth and development of perilla. The main environmental factor data in six ecological area in Guizhou province were collected, and the correlation analysis with yield and quality traits of 15 perilla strains was conducted. The results showed that the cultivation environment has significant effects on the yield and quality traits of perilla. The effect of environment on main yield composed traits, contained grain number in top spike, effective panicle number per plant, plant height, top spike length, growth period, and thousand seed was degressive. In the different environmental factors, the latitude showed positive correlation with yield, growth period and effective panicle number per plant, and negative correlation with top spike length and grain number in top spike. Elevation showed negative correlation with the growth period of perilla. The perilla yield increased at first and then decreased with altitude rising, with the maximum in the 800 m altitude. The 600-900 m altitude is suitable area for perilla. Except for positive correlation with the plant height, and negative correlation with top spike length, the longitude showed in apparent impact on other traits. Sunshine duration, temperature and rainfall accumulation showed different effect on the different perilla strains. For yield composed traits, the sunshine duration was negatively correlation with the plant length. The accumulated temperature and mean temperature showed negative correlation with the main spike length, the rainfall showed negative correlation with the precipitation and growth period, plant height, ear number. The environmental impact on the oil compounds decreased with oleic , stearic , linoleic , -linolenic , and oil content. Correlation analysis showed that the significantly negative correlation between the oil content and and linoleic content, and the positive correlation between linolenic content, -linolenic content showed significant negative correlation with other fatty acids composition, and , stearic , oleic , linoleic showed significant positive correlation with each other. The influence of different environmental factors on the quality of perilla were as follows: the oil content was positively associated with elevation and sunshine duration. -Linolenic content showed negative correlation with longitude, latitude, accumulated temperature and mean temperature, but positive correlation with altitude, sunlight and rainfall capacity. The correlation between , stearic , oleic , linoleic and environmental factors showed contrast character of -linolenic . This study detailed discussed the influence of environmental factors on the quality of perilla, which provided the foundation of ecological planting technology and geoherbalism research of perilla.Copyright© by the Chinese Pharmaceutical Association.

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Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering hyperlipidemia via regulating the AMPK/SREBP1/PPARα signaling cascade of lipid metabolism.

The present study for the first time aims to examine the hypothesis that circulating gamma-glutamyl carboxylated growth arrest specific protein 6 (Gla-Gas6) deficiency may be associated with hyperlipidemia and vitamin K (VK) supplementation may ameliorate the impaired lipid homeostasis via activating Gas6 protein. Subjects with hyperlipidemia (n=22) and age-matched healthy controls (n=19) were included in this study. Results showed that plasma levels of Gla-Gas6 protein and VK were significantly lower in hyperlipidemic subjects compared to control. Moreover, Gla-Gas6 levels were significantly and positively correlated with VK (P=.034, r=0.452) and negatively with triglyceride (P=.022, r=-0.485) and total cholesterol (P=.043, r=-0.435) in hyperlipidemic subjects, which suggest that VK supplementation may have a positive effect in activating Gas6 protein and thereby reducing the aberrant plasma lipid levels. Further studies with high-fat diet (HFD)-fed animal model of hyperlipidemia demonstrated that VK supplementation (5 μg/kg , 8 weeks) reduced the plasma lipid levels, stimulated both the plasma levels and the hepatic protein expression of Gla-Gas6 protein, and regulated the AMPK/SREBP1/PPARα signaling pathways of hepatic lipid metabolism in HFD-fed mice. Moreover, by using (PA, 0.75 mM)-treated both control and GGCX knockdown hepatocytes, this study dissected the direct role of Gla-Gas6 in mediating the positive effect of VK on preventing the PA-induced impaired hepatic lipid metabolism via regulating AMPK/SREBP1/PPARα pathways. Combining all, the present study demonstrated the beneficial effect of VK supplementation in preventing the impaired lipid homeostasis via activating VK-dependent Gas6 protein.Copyright © 2019 Elsevier Inc. All rights reserved.

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Macrophage alternative activation confers protection against lipotoxicity-induced cell death.

Alternative activation (M2) of adipose tissue resident macrophage (ATM) inhibits obesity-induced metabolic inflammation. The underlying mechanisms remain unclear. Recent studies have shown that dysregulated lipid homeostasis caused by increased lipolysis in white adipose tissue (WAT) in the obese state is a trigger of inflammatory responses. We investigated the role of M2 macrophages in lipotoxicity-induced inflammation.We used microarray experiments to profile macrophage gene expression regulated by two M2 inducers, interleukin-4 (Il-4), and peroxisome proliferator-activated receptor delta/gamma (Pparδ/Pparγ) agonists. Functional validation studies were performed in bone marrow-derived macrophages and mice deprived of the signal transducer and activator of transcription 6 gene (Stat6; downstream effector of Il-4) or Pparδ/Pparγ genes (downstream effectors of Stat6). (PA) and β-adrenergic agonist were employed to induce macrophage lipid loading in\xa0vitro and in\xa0vivo, respectively.Profiling of genes regulated by Il-4 or Pparδ/Pparγ agonists reveals that alternative activation promotes the cell survival program, while inhibiting that of inflammation-related cell death. Deletion of Stat6 or Pparδ/Pparγ increases the susceptibility of macrophages to PA-induced cell death. NLR family pyrin domain containing 3 (Nlrp3) inflammasome activation by PA in the presence of lipopolysaccharide is also increased in Stat6 macrophages and to a lesser extent, in Pparδ/γ macrophages. In concert, β-adrenergic agonist-induced lipolysis results in higher levels of cell death and inflammatory markers in ATMs derived from myeloid-specific Pparδ/γ or Stat6 mice.Our data suggest that ATM cell death is closely linked to metabolic inflammation. Within WAT where concentrations of free fatty acids fluctuate, M2 polarization regulated by the Stat6-Ppar axis enhances ATM\'s tolerance to lipid-mediated stress, thereby maintaining the homeostatic state.Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

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Evaluation of bioactive compounds in black table olives fermented with selected microbial starters.

Table olives have been a component of the Mediterranean diet for centuries, with the trend for their consumption currently increasing worldwide. They are rich in bioactive molecules with nutritional, antioxidant, anti-inflammatory or hormone-like properties. In the present study, the concentrations of phenolics, triterpenic acids, carotenoids and vitamins, as well as fatty profiles and antioxidant activity, were analyzed in the edible portion of black table olives (Olea europea L.) from Italian (Cellina di Nardò and Leccino) and Greek (Kalamàta and Conservolea) cultivars fermented with selected autochthonous starters and in the corresponding monovarietal olive oils.On a fresh basis, Cellina di Nardò and Leccino table olives showed the highest total phenolic content. No significant differences were found with respect to the levels of total triterpenic (maslinic and oleanolic) acids and vitamin E among cultivars. All table olives were characterized by high amounts of oleic, linoleic and acids. Oils were richer in lipophilic antioxidants (carotenoids and tocochromanols) than table olives, which, instead, showed a higher content of polyphenols and triterpenic acids than oils.The present study demonstrates that fermented table olives are an excellent natural source of unsaturated fatty acids, as well as being nutritionally important health-promoting bioactive compounds. © 2017 Society of Chemical Industry.© 2017 Society of Chemical Industry.

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Orange juice affects acylcarnitine metabolism in healthy volunteers as revealed by a mass-spectrometry based metabolomics approach.

Citrus juices, especially orange juice, constitute rich sources of bioactive compounds with a wide range of health-promoting activities. Data from epidemiological and in vitro studies suggest that orange juice (OJ) may have a positive impact on lipid metabolism. However, the effect of orange juice intake on blood lipid profile is still poorly understood. We have used two different blood samples, Dried Blood Spots (DBS) and plasma, to assess the effect of two-week orange juice consumption in healthy volunteers by a mass-spectrometry based metabolomics approach. DBS were analysed by liquid chromatography mass spectrometry (LC-MS) and plasma samples were analysed by the gas chromatography mass spectrometry (GC-MS). One hundred sixty-nine lipids including acylcarnitines (AC), lysophosphatidylcholines (LysoPC), (diacyl- and acyl-alkyl-) phosphatidylcholines (PC aa and PC ae) and sphingomyelins (SM) were identified and quantified in DBS. Eighteen fatty acids were identified and quantified in plasma. Multivariate analysis allowed to identify an increase in C3:1, C5-DC(C6-OH), C5-M-DC, C5:1-DC, C8, C12-DC, lysoPC18:3, myristic , pentadecanoic , palmitoleic and and a decrease in nervonic , C0, C2, C10, C10:1, C16:1, C16-OH, C16:1-OH, C18-OH, PC aa C40:4, PC ae C38:4, PC ae C42:3, PC ae C42:4 and cholesterol levels after orange juice intake. A two-week period of orange juice intake could affect fatty acids β-oxidation through mitochondrial and peroxisomal pathways, leading to an increase of short-chain acylcarnitines and a decrease of medium and long-chain acylcarnitines. This is the first report analyzing the effect of orange juice intake in healthy volunteers using a dried blood spot-based metabolomics approach.Copyright © 2018 Elsevier Ltd. All rights reserved.

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Nutritional modulation of metabolic inflammation.

Metabolic inflammation is a very topical area of research, wherein aberrations in metabolic and inflammatory pathways probably contribute to atherosclerosis, insulin resistance (IR) and type 2 diabetes. Metabolic insults arising from obesity promote inflammation, which in turn impedes insulin signalling and reverse cholesterol transport (RCT). Key cells in the process are metabolically activated macrophages, which up-regulate both pro- and anti-inflammatory pathways in response to lipid spillover from adipocytes. Peroxisome proliferator-activated receptors and AMP-activated protein kinase (AMPK) are regulators of cellular homeostasis that influence both inflammatory and metabolic pathways. Dietary fats, such as saturated fatty acids (SFAs), can differentially modulate metabolic inflammation. , in particular, is a well-characterized nutrient that promotes metabolic inflammation via the NLRP3 (the nod-like receptor containing a pyrin domain) inflammasome, which is partly attributable to AMPK inhibition. Conversely, some unsaturated fatty acids are less potent agonists of metabolic inflammation. For example, monounsaturated fatty does not reduce AMPK as potently as SFA and n-3 polyunsaturated fatty acids actively resolve inflammation via resolvins and protectins. Nevertheless, the full extent to which nutritional state modulates metabolic inflammation requires greater clarification.© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

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Serine prevented high-fat diet-induced oxidative stress by activating AMPK and epigenetically modulating the expression of glutathione synthesis-related genes.

Serine deficiency has been observed in patients with nonalcoholic fatty liver disease (NAFLD). Whether serine supplementation has any beneficial effects on the prevention of NAFLD remains unknown. The present study was conducted to investigate the effects of serine supplementation on hepatic oxidative stress and steatosis and its related mechanisms. Forty male C57BL/6J mice (9week-old) were randomly assigned into four groups (n=10) and fed: i) a low-fat diet; ii) a low-fat diet supplemented with 1% (wt:vol) serine; iii) a high-fat (HF) diet; and iv) a HF diet supplemented with 1% serine, respectively. (PA)-treated primary hepatocytes separated from adult mice were also used to study the effects of serine on oxidative stress. The results showed that serine supplementation increased glucose tolerance and insulin sensitivity, and protected mice from hepatic lipid accumulation, but did not significantly decreased HF diet-induced gain. In addition, serine supplementation protected glutathione (GSH) antioxidant system and prevented hypermethylation in the promoters of glutathione synthesis-related genes, while decreasing reactive oxygen species (ROS) in mice fed a HF diet. Moreover, we found that serine supplementation increased phosphorylation and S-glutathionylation of AMP-activated protein kinase α subunit (AMPKα), and decreased ROS, malondialdehyde and triglyceride contents in PA-treated primary hepatocytes. However, while AMPK activity or GSH synthesis was inhibited, the abovementioned effects of serine on PA-treated primary hepatocytes were not observed. Our results suggest that serine supplementation could prevent HF diet-induced oxidative stress and steatosis by epigenetically modulating the expression of glutathione synthesis-related genes and through AMPK activation.Copyright © 2017 Elsevier B.V. All rights reserved.

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Antibacterial, Anti-Diarrhoeal, Analgesic, Cytotoxic Activities, and GC-MS Profiling of (Buch.-Ham.) Seed.

Fruits of (Buch.-Ham.), (English: mangrove apple, Bengali: keora) both seeds and pericarps, are largely consumed as food besides their enormous medicinal application. The fruit seeds have high content of nutrients and bioactive components. The seeds powder of was successively fractionated using -hexane, diethyl ether, chloroform, ethyl acetate, and methanol. The fractions were used to evaluate antibacterial, anti-diarrhoeal, analgesic, and cytotoxic activities. Methanol fraction of seeds (MeS) stronly inhibited strains, Paratyphi A, Typhi, , and except at 500 μg/disc. All the fractions strongly inhibited castor oil induced diarrhoeal episodes and onset time in mice at 500 mg extract/kg (<0.001). At the same concentration, MeS had the strongest inhibitory activity on diarrhoeal episodes, whereas the -hexane fraction (HS) significantly (<0.05) prolonged diarrhoeal onset time as compared to positive control. Similarly, HS (<0.005) inhibited acetic induced writhing in mice at 500 mg extract/kg, more than any other fraction. HS and diethyl ether fractions of seed strongly increased reaction time of mice in hot plate test at 500 mg extract/kg. All the fractions showed strong cytotoxic effects in brine shrimp lethality tests. Gas chromatography-mass spectrometry analysis of HS led to the identification of 23 compounds. Linoleic (29.9%), (23.2%), ascorbyl palmitate (21.2%), and stearic (10.5%) were the major compounds in HS. These results suggest that seeds of could be of great use as nutraceuticals.

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Chemical Composition of Pinus roxburghii Bark Volatile Oil and Validation of Its Anti-Inflammatory Activity Using Molecular Modelling and Bleomycin-Induced Inflammation in Albino Mice.

The chemical composition of bark essential oil (PRO) was qualitatively and quantitatively determined using GC/FID and GC/MS. The anti-inflammatory activity was assessed in vitro by evaluating the binding percentages on the cannabinoids and opioids receptors. Bleomycin (BLM)-induced pulmonary inflammation in mice was adopted to assess PRO anti-inflammatory efficacy in vivo. In silico molecular modelling of its major components was performed on human glucocorticoids receptor (GR). Seventy-five components were identified in which longifolene (33.13%) and (9.34%) constituted the predominant components. No binding was observed on cannabinoid receptor type 1 (CB1), whereas mild binding was observed on cannabinoid receptor type 2 (CB2), , , and receptors accounting for 2.9%, 6.9%, 10.9% and 22% binding. A significant in vivo activity was evidenced by reduction of the elevated malondialdehyde (MDA), nitric oxide (NO), myeloperoxidase (MPO), interleukin-6 (IL-6), and tumor necrosis factor- (TNF-) levels by 55.56%, 55.66%, 64.64%, 58.85% and 77.78% with concomitant elevation of superoxide dismutase (SOD) and catalase (CAT) activities comparable to BLM-treated group at 100 mg/kg . In silico studies showed that exerted the fittest binding. PRO could serve as a potent anti-inflammatory natural candidate that should be supported by further clinical trials.

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Saturated fatty acids, obesity, and the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in asthmatic patients.

Both obesity and high dietary fat intake activate the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome.We aimed to examine NLRP3 inflammasome activity in the airways of obese asthmatic patients after macronutrient overload and in immune cells challenged by inflammasome triggers.Study 1 was a cross-sectional observational study of nonobese (n\xa0=\xa051) and obese (n\xa0=\xa076) asthmatic adults. Study 2 was a randomized, crossover, acute feeding study in 23 asthmatic adults (n\xa0=\xa012 nonobese and n\xa0=\xa011 obese subjects). Subjects consumed 3 isocaloric meals on 3 separate occasions (ie, saturated fatty , n-6 polyunsaturated fatty , and carbohydrate) and were assessed at 0 and 4\xa0hours. For Studies 1 and 2, airway inflammation was measured based on sputum differential cell counts, IL-1β protein levels (ELISA), and sputum cell gene expression (Nanostring nCounter). In Study 3 peripheral blood neutrophils and monocytes were isolated by using Ficoll density gradient and magnetic bead separation and incubated with or without , LPS, or TNF-α for 24\xa0hours, and IL-1β release was measured (ELISA).In Study 1 NLRP3 and nucleotide oligomerization domain 1 (NOD1) gene expression was upregulated, and sputum IL-1β protein levels were greater in obese versus nonobese asthmatic patients. In Study 2 the saturated fatty meal led to increases in sputum neutrophil percentages and sputum cell gene expression of Toll-like receptor 4 (TLR4) and NLRP3 at 4\xa0hours in nonobese asthmatic patients. In Study 3 neutrophils and monocytes released IL-1β when challenged with a combination of and LPS or TNF-α.The NLRP3 inflammasome is a potential therapeutic target in asthmatic patients. Behavioral interventions that reduce fatty exposure, such as loss and dietary saturated fat restriction, warrant further exploration.Copyright © 2018 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

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Human relaxin-2 attenuates hepatic steatosis and fibrosis in mice with non-alcoholic fatty liver disease.

Human relaxin-2 reduces hepatic fibrosis in mice. However, the effects of relaxin-2 on hepatic steatosis and fibrosis in animals with non-alcoholic fatty liver disease (NAFLD) remain to be elucidated. C57BL/6 mice fed a high-fat diet (HFD) or methionine-choline-deficient (MCD) diet were randomly assigned to receive recombinant human relaxin-2 (25 or 75\u2009μg/kg/day) or vehicle for 4 weeks. In HFD-fed mice, relaxin-2 decreased systemic insulin resistance and reduced , epididymal fat mass and serum leptin and insulin concentrations. In livers of HFD-fed mice, relaxin-2 attenuated steatosis and increased phosphorylation of insulin receptor substrate-1, Akt and endothelial nitric oxide synthase (eNOS), and activated genes that regulate fatty oxidation and suppressed acetyl-CoA carboxylase. Relaxin-2 had no direct anti-steatotic effect on primary mouse hepatocytes, but S-nitroso-N-acetylpenicillamine attenuated -induced steatosis and activated genes regulating fatty oxidation in hepatocytes. In mice fed an MCD diet, relaxin-2 attenuated steatosis, inflammation and fibrosis. Relaxin-2 increased eNOS and Akt phosphorylation and transcript levels of cytochrome P450-4a10 and decreased acetyl-CoA carboxylase in MCD-fed mouse livers. Moreover, expression levels of Kupffer cell activation, hepatic stellate cell activation and hepatocyte apoptosis were decreased in MCD diet-fed mice receiving relaxin-2. In conclusion, relaxin-2 reduces hepatic steatosis by activating intrahepatic eNOS in HFD-fed mice and further attenuates liver fibrosis in MCD diet-fed mice. Therefore, human relaxin-2 is a potential therapeutic treatment for NAFLD.

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Increased serum concentration of ceramides in obese children with nonalcoholic fatty liver disease.

Hepatic lipid accumulation is closely related to the development of insulin resistance, which is regarded as one of the most significant risk factors of nonalcoholic fatty liver disease (NAFLD). Although the exact molecular pathway leading to impaired insulin signaling has not been definitively established, ceramides are suspected mediators of lipid induced hepatic insulin resistance. Therefore, the aim of the study was to evaluate the serum ceramides concentration in obese children with NAFLD.The prospective study included 80 obese children (aged 7-17\xa0years, median 12\xa0years) admitted to our Department to diagnose initially suspected liver disease. Patients with viral hepatitis (HCV, HBV, CMV), autoimmune (AIH), toxic and metabolic (Wilson\'s disease, alfa-1-antitrypsin deficiency) liver diseases and celiac disease were excluded. NAFLD was diagnosed based on pediatric diagnostic criteria in obese children with liver steatosis in ultrasound (US) as well as elevated alanine transaminase (ALT) serum activity after exclusion of other major liver diseases listed before. Ultrasonography was used as a screening method and for qualitative assessment of the steatosis degree (graded according to Saverymuttu scale). Advanced steatosis was defined as a score\u2009>\u20091. The total intrahepatic lipid content (TILC) was assessed by magnetic resonance proton spectroscopy (HMRS) which is the most accurate technique for assessment of ectopic fat accumulation. Fasting serum concentration of ceramides was measured in 62 children.NAFLD was diagnosed in 31 children. Significant, positive correlation was found between total serum concentration of ceramides and insulin (r\xa0=\u20090.3, p\xa0=\u20090.02) and HOMA-IR (r\xa0=\u20090.28, p\xa0=\u20090.03). Total ceramide concentration as well as specific fatty -ceramides (FA-ceramides) concentrations, namely: myristic, , palmitoleic, stearic, oleic, behenic and lignoceric were significantly higher (p\xa0=\u20090.004, p\xa0=\u20090.003, p\xa0=\u20090.007, p\xa0<\u20090.001, p\xa0=\u20090.035, p\xa0=\u20090.008, p\xa0=\u20090.003, p\xa0=\u20090.006, respectively) in children with NAFLD compared to controls (n\xa0=\u200914). Moreover, children with NAFLD had significantly higher activity of ALT (p\xa0<\u20090.001) and GGT (p\xa0<\u20090.001), HOMA-IR (p\xa0=\u20090.04), BMI (p\xa0=\u20090.046), waist circumference (p\xa0=\u20090.01) steatosis grade in ultrasound (p\xa0<\u20090.001) and TILC in HMRS (p\xa0<\u20090.001) compared to children without NAFLD. We did not find significant differences in total and FA-ceramide species concentrations between children with mild (grade 1) and advanced liver steatosis in ultrasonography (grade 2-3).Elevated ceramide concentrations in obese patients together with their significant correlation with insulin resistance parameters suggest their association with molecular pathways involved in insulin signaling impairment known to be strongly linked to pathogenesis of non-alcoholic fatty liver disease.

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A Combination of Short and Simple Surfactant Protein B and C Analogues as a New Synthetic Surfactant: In Vitro and Animal Experiments.

Pulmonary surfactants for preterm infants contain mostly animal-derived surfactant proteins (SPs), which are essential for lowering surface tension. We prepared artificial pulmonary surfactants using synthetic human SP analogs and performed in vitro and in vivo experiments.We synthesized peptide analogues that resemble human SP-B (RMLPQLVCRLVLRCSMD) and SP-C (CPVHLKRLLLLLLLLLLLLLLLL). Dipalmitoylphosphatidylcholine (DPPC), phosphatidylglycerol (PG), and (PA) were added and mixed in lyophilized to render powdered surfactant. Synsurf-1 was composed of DPPC:PG:PA:SP-B (75:25:10:3, w/w); Synsurf-2 was composed of DPPC:PG:PA:SP-C (75:25:10:3, w/w); and Synsurf-3 was composed of DPPC:PG:PA:SP-B:SP-C (75:25:10:3:3, w/w). We performed in vitro study to compare the physical characteristics using pulsating bubble surfactometer and modified Wilhelmy balance test. Surface spreading and adsorption test of the surfactant preparations were measured. In vivo test was performed using term and preterm rabbit pups. Pressure-volume curves were generated during the deflation phase. Histologic findings were examined.Pulsating bubble surfactometer readings revealed following minimum and maximum surface tension (mN/m) at 5 minutes: Surfacten® (5.5±0.4, 32.8±1.6), Synsurf-1 (16.7±0.6, 28.7±1.5), Synsurf-2 (7.9±1.0, 33.1±1.6), and Synsurf-3 (7.1±0.8, 34.5±1.0). Surface spreading rates were as follows: Surfacten® (27 mN/m), Synsurf-1 (43 mN/m), Synsurf-2 (27 mN/m), and Synsurf-3 (27 mN/m). Surface adsorption rate results were as follows: Surfacten® (28 mN/m), Synsurf-1 (35 mN/m), Synsurf-2 (29 mN/m), and Synsurf-3 (27 mN/m). The deflation curves were best for Synsurf-3; those for Synsurf-2 were better than those for Surfacten®. Synsurf-1 was the worst surfactant preparation. Microscopic examination showed the largest aerated area of the alveoli in the Synsurf-3 group, followed by Synsurf-1 and Surfacten®; Synsurf-2 was the smallest.Synsurf-3 containing both SP-B and SP-C synthetic analogs showed comparable and better efficacy than commercially used Surfacten® in lowering surface tension, pressure-volume curves, and tissue aerated area of the alveoli.© Copyright: Yonsei University College of Medicine 2017

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Melatonin improves insulin resistance and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with insulin resistance and type 2 diabetes. The aim of this study was to determine whether melatonin improves hepatic insulin resistance and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve insulin resistance in FETUA-overexpressing cells, indicating that improvement in insulin resistance by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced gain to one-third of that of the HFD group and hepatic steatosis. Insulin sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the liver and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic insulin resistance and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

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Chronic intake of moderate fat-enriched diet induces fatty liver and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for , biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

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Vasoreactivity of isolated aortic rings from dyslipidemic and insulin resistant inducible nitric oxide synthase knockout mice.

Recent study from this lab indicated enhanced susceptibility of iNOS KO mice for diet induced obesity (DIO) and systemic insulin resistance (IR) as compared to C57BL/6 (WT) mice. The present study investigates aortic vasoreactivity in high fat diet (HFD) induced insulin resistant iNOS KO mice. WT and iNOS KO mice were fed with 45% HFD/10% LFD for ten weeks. Systemic IR was assessed via measurement of circulating lipids, glucose, and insulin; while phenylephrine (PE)/acetylcholine (ACh) induced responses were monitored in the isolated aortic rings. To understand the mechanism, qPCR or Western blotting experiments were performed in aorta and Ea.hy926\u202fcells. After 10 weeks of HFD feeding, significant increase in the /fat mass, augmented circulating lipids, glucose, insulin and inflammatory cytokines along with impaired acetylcholine induced aortic vasorelaxation and enhanced iNOS expression was observed in the aortic tissue of WT mice. In the aminoguanidine (AG, 20\u202fmg/kg for 4 weeks) treated WT mice and also in iNOS KO mice, acetylcholine induced vasorelaxation was significantly preserved. Further, acetylcholine mediated vasorelaxation correlated with increased eNOS phosphorylation at Ser1177 residue in the iNOS KO mice and same was also observed in the iNOS silenced Ea.hy926\u202fcells. Moreover, treatment of Ea.hy926\u202fcells with or TNFα also caused a significant decrease in eNOS activity, which was reversed in iNOS silenced Ea.hy926\u202fcells suggesting the role of iNOS in the reduction of eNOS activity. The study thus implies a critical role of iNOS in vascular diseases associated with dyslipidemia/IR.Copyright © 2019. Published by Elsevier B.V.

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A new lipid carrier protein in the cattle tick Rhipicephalus microplus.

Tick infestation in cattle reflects the main cause of economic loss to cattle producers. This is due to several reasons but mainly to their ability to feed on blood and generate a huge amount of eggs. Lipid transport in arthropods is achieved by highly specialized hemolymphatic lipoproteins, which resemble those described in vertebrate blood. Such lipoproteins continuously deliver lipids through the blood to growing eggs. The injection of radioactive [H] into tick hemocoel showed that the gut, ovary, fat and Gene\'s organ were the main organs of incorporation of this labeled fatty . The rate of [H] incorporation by the organs was high up to 30\u202fmin after injection. The [H] incorporated by these organs was later found in phospholipids and neutral lipids. Here, we describe the purification and characterization of a key player of lipid dynamics in tick hemolymph. The Rhipicephalus microplus lipid-apolipoprotein complex (RmLCP) is a new high-density lipoprotein (1.18\u202fg/mL), which accounts for over 90% of [H] present in the hemolymph. It has a native molecular of 420\u202fkDa and is composed of one subunit of 122\u202fkDa. Protein identification analysis of RmLPC subunit showed two better hits: vitellogenin 2 (23% protein coverage) and vitellogenin 5 (29% protein coverage), respectively and similarities with hemolymphatic apolipoproteins of arachnids such as the tick Ixodes scapularis (80%), the mite Galendromus occidentalis (44%) and the spider Parasteatoda tepidariorum (43%) and also for the insects Locusta migratoria (45%), Drosophila melanogaster (42%) and Manduca sexta (47%) to vitellogenin 2 and tick Ixodes scapularis (83%), the crab Limulus polyphemus (55%) and the oyster Crassostrea gigas (55%) to vitellogenin 5. Furthermore, it shows a distinct lipid composition from most arthropod lipoproteins, being composed of 40% free cholesterol, 27% phospholipids, 20% triacylglycerol and 15% hydrocarbons. In addition to binding most hemolymphatic fatty acids, this lipoprotein also binds and transports free cholesterol. In conclusion, the present study provides insight into the macromolecules involved in arachnid metabolism, which have significant potential for future use for the biological control of ticks.Copyright © 2018 Elsevier GmbH. All rights reserved.

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Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

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Activity-Structure Study on the Peptide Fraction of AG2: a Potent In Vitro Transfection Agent.

Gemini-based amphiphiles are candidates for biomedical applications. In fact, most of the gemini compounds described in the literature have been prepared to be used as new synthetic vectors in gene transfection. Our group carried out an activity-structure study starting from the structure of the gemini [AG2-C], which is an effective in vitro transfection reagent. We synthesized a series of novel amphiphilic amino derivatives of low molecular , named AGn-C (N), in which the same apolar region (m) of oleic or was maintained and the peptide region was modified by amino insertions, deletions, and substitutions. We also determined the transfection efficiency, critical micelle concentration, particle size, and ζ-potential for these derivatives. Amphiphiles AG10-C and AG10-C were more active at a lower N/P ratio than AG2-C. These amphiphiles showed no activity when lysine was replaced by ornithine, and the activity of all derivatives increased when there were more ornithine residues and a W/O\u2009=\u20091 ratio in the peptide region. It can be said that for AG10-C, these two structural requirements on the amino portion predominated over the type of aliphatic chain used.

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Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high-fat diet-fed mice.

Type 2 diabetes mellitus is frequently accompanied by fatty liver disease. Lipid accumulation within the liver is considered as one of the risk factors for insulin resistance. Hepatocyte growth factor (HGF) is used to treat liver dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting insulin and homeostatic model assessment of insulin resistance were calculated for insulin sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/fatty synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic insulin resistance and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.

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Blockade of myeloid differentiation protein 2 prevents obesity-induced inflammation and nephropathy.

Obesity is a major and independent risk factor of kidney diseases. The pathogenic mechanisms of obesity-associated renal injury are recognized to at least involve a lipid-rich and pro-inflammatory state of the renal tissues, but specific mechanisms establishing causal relation remain unknown. Saturated fatty acids are elevated in obesity, and known to induce chronic inflammation in kidneys. Myeloid differentiation protein 2 (MD2) is an important protein in lipopolysaccharide-induced innate immunity response and inflammation. We suggested that obesity-associated renal injury is regulated by MD2 thereby driving an inflammatory renal injury. The used three mouse models for in vivo study: MD2 knockout mice (KO) maintained on high fat diet (HFD), wild-type mice on HFD plus L6H21, a specific MD2 inhibitor and KO mice given (PA) by IV injection. The in vitro studies were carried out in cultured renal tubular epithelial cells, mouse mesangial cells and primary macrophages, respectively. The HFD mice presented with increased hyperlipidemia, serum creatinine and proteinuria. Renal tissue from HFD mice had increased fibrosis, inflammatory cytokines, macrophage infiltration, and activation of NF-κB and MAPKs. This HFD-induced renal injury profile was not observed in KO mice or L6H21-treated mice. Mice given PA mimmicked the HFD-induced renal injury profiles, which were prevented by MD2 knockout. The in vitro data further confirmed MD2 mediates PA-induced inflammation. MD2 is causally related with obesity-associated renal inflammatory injury. We believe that MD2 is an attractive target for future therapeutic strategies in obesity-associated kidney diseases.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

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Polysaccharide from Mulberry Fruit ( L.) Protects against -Induced Hepatocyte Lipotoxicity by Activating the Nrf2/ARE Signaling Pathway.

This study was aimed to investigate the protective effects of three different mulberry fruit polysaccharide fractions (MFP-I, MFP-II, and MFP-III) against (PA)-induced hepatocyte lipotoxicity and characterize the functional polysaccharide fraction using gel permeation chromatography, high-performance liquid chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance analyses. MFP-I, MFP-II, and MFP-III were isolated from mulberry fruit by stepwise precipitation with 30, 60, and 90% ethanol, respectively. MFP-II at 0.1 and 0.2 mg/mL dramatically attenuated PA-induced hepatic lipotoxicity, while MFP-I and MFP-III showed weak protection. It was demonstrated that MFP-II not only increased nuclear factor erythroid-2-related factor 2 (Nrf2) phosphorylation and its nuclear translocation, thereby activating the Nrf2/ARE signaling pathway, but also enhanced heme oxygenase 1, NAD(P)H:quinone oxidoreductase 1, and γ-glutamate cysteine ligase gene expressions and promoted catalase and glutathione peroxidase activities, which protected hepatocytes against PA-induced oxidative stress and lipotoxicity. Further investigation indicated that the molecular of MFP-II was 115.0 kDa, and MFP-II mainly consisted of galactose (30.5%), arabinose (26.2%), and rhamnose (23.1%). Overall, our research might provide in-depth insight into mulberry fruit polysaccharide in ameliorating lipid metabolic disorders.

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Interesterified Palm Olein (IEPalm) and Interesterified Stearic -Rich Fat Blend (IEStear) Have No Adverse Effects on Insulin Resistance: A Randomized Control Trial.

Chemically-interesterified (CIE) fats are -fat free and are increasingly being used as an alternative to hydrogenated oils for food manufacturing industries to optimize their products\' characteristics and nutrient compositions. The metabolic effects of CIE fats on insulin activity, lipids, and adiposity in humans are not well established. We investigated the effects of CIE fats rich in (C16:0, IEPalm) and stearic (C18:0, IEStear) acids on insulin resistance, serum lipids, apolipoprotein concentrations, and adiposity, using C16:0-rich natural palm olein (NatPO) as the control. We designed a parallel, double-blind clinical trial. Three test fats were used to prepare daily snacks for consumption with a standard background diet over a period of 8 weeks by three groups of a total of 85 healthy, overweight adult volunteers. We measured the outcome variables at weeks 0, 6, and at the endpoint of 8. After 8 weeks, there was no significant difference in surrogate biomarkers of insulin resistance in any of the IE fat diets (IEPalm and IEStear) compared to the NatPO diet. The change in serum triacylglycerol concentrations was significantly lower with the IEStear diet, and the changes in serum leptin and fat percentages were significantly lower in the NatPO-diet compared to the IEPalm diet. We conclude that diets containing C16:0 and C18:0-rich CIE fats do not affect markers of insulin resistance compared to a natural C16:0-rich fat (NatPO) diet. Higher amounts of saturated fatty acids (SFAs) and longer chain SFAs situated at the -1,3 position of the triacylglycerol (TAG) backbones resulted in less gain and lower changes in fat percentage and leptin concentration to those observed in NatPO and IEStear.

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MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue inflammation in obesity. Both MIP-1α and free fatty acids are elevated in obesity/T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling pathways were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling pathways. In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in obesity/T2D may contribute to metabolic inflammation through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

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Optimization of Synechococcus sp. VDW Cultivation with Artificially Prepared Shrimp Wastewater for Ammonium Removal and Its Potential for Use As a Biofuel Feedstock.

To investigate the potential of application of marine cyanobacterium for concurrent biomass production and ammonium removal, Synechococcus sp. VDW was cultured under different conditions in medium containing varying concentrations of NHCl. Response surface methodology (RSM) was then used to build a predictive model of the combined effects of independent variables (pH, inoculum size, ammonium concentration). At the optimum conditions of initial pH 7.4, inoculum size 0.17 (OD730) and ammonium concentration 10.5 mg L, the maximum ammonium removal and biomass productivity were about 95% and 34 mg Ld, respectively, after seven days of cultivation. The result of fatty methyl ester (FAME) analysis showed that the major fatty acids were (C16:0), linoleic (C18:2 n6 cis), palmitoleic (C16:1) and oleic (C18:1 n9 cis), which accounted for more than 80% of total fatty acids. Further, analysis of neutral lipid accumulation using flow cytometry revealed that the mean of the fluorescence intensity increased under optimal conditions. These results indicate that Synechococcus sp. VDW has the potential for use for concurrent water treatment and production of biomass that can be applied as biofuel feedstock.

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Effects of timing of supplementation on production responses of early-lactation dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a diet supplemented with C16:0 (, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% fat-corrected milk by 5.30 kg/d, yield of energy-corrected milk (ECM) by 4.70 kg/d, milk fat content by 0.41% units, milk fat yield by 280 g/d, and protein yield by 100 g/d. The increase in milk fat associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased (BW) by 21 kg and condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk fat yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% fat-corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk fat content by 0.22% units, milk fat yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk fat yield due to feeding PA during the PK period increasing milk fat yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High-Fat-Diet-Induced Cardiac Dysfunction and Insulin Resistance by Elevating Tafazzin in Mouse Hearts.

High fat (HF)-diet-induced insulin resistance is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac insulin signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low fat (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs insulin signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the mitochondria. Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores insulin sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced insulin resistance further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to insulin resistance via mediating Drp-1 translocation to the mitochondria, and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates insulin resistance.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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Tannic , a novel histone acetyltransferase inhibitor, prevents non-alcoholic fatty liver disease both in\xa0vivo and in\xa0vitro model.

We examined the potential of tannic (TA) as a novel histone acetyltransferase inhibitor (HATi) and demonstrated that TA prevents non-alcoholic fatty liver disease (NAFLD) by inhibiting HAT activity.The anti-HAT activity of TA was examined using HAT activity assays. An in\xa0vitro NAFLD model was generated by treating HepG2 cells with oleic and acids. Male C57BL/6J mice were fed a control diet (CD) or Western diet (WD) with or without supplementation with either 1% or 3% TA (w/w) for 12 weeks. Finally, the possibility of interacting p300 and TA was simulated.TA suppressed HAT activity both in\xa0vitro and in\xa0vivo. Interestingly, TA abrogated occupancy of p300 on the sterol regulatory element in the fatty synthase and ATP-citrate lyase promoters, eventually inducing hypoacetylation of H3K9 and H3K36. Furthermore, TA decreased acetylation at lysine residues 9 and 36 of histone H3 protein and that of total proteins. Consequently, TA decreased the mRNA expression of lipogenesis-related genes and attenuated lipid accumulation in\xa0vivo. We observed that NAFLD features, including , liver mass, fat mass, and lipid profile in serum, were improved by TA supplementation in\xa0vivo. Finally, we demonstrated the possibility that TA directly binds to p300 through docking simulation between ligand and protein.Our findings demonstrate that TA, a novel HATi, has potential application for the prevention of NAFLD.Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.

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Chronic administration of myristic improves hyperglycaemia in the Nagoya-Shibata-Yasuda mouse model of congenital type 2 diabetes.

Previously, we demonstrated that myristic (14:0) increases levels of diacylglycerol kinase (DGK) δ, a key enzyme involved in type 2 diabetes exacerbation, and enhances glucose uptake in C2C12 myotube cells. Moreover, results from a population-based cohort study suggest that consumption of high-fat dairy products, which contain high amounts of myristic , is associated with a lower risk of developing type 2 diabetes. Taken together, we hypothesised that intake of myristic reduces type 2 diabetes risk in vivo. The aim of this study was to examine the glucose-lowering effect of myristic in Nagoya-Shibata-Yasuda (NSY) mice, a spontaneous model for studying obesity-related type 2 diabetes.Male NSY mice were orally administered vehicle (n\xa0=\xa09), 300\xa0mg/kg of myristic (n\xa0=\xa014) or 300\xa0mg/kg of (16:0) (n\xa0=\xa09) every other day from 4\xa0weeks of age. Glucose and insulin tolerance tests were performed at weeks 18, 24 and 30, and weeks 20 and 26, respectively. DGKδ levels were measured in skeletal muscle from 32-36-week-old NSY mice via western blot.Chronic oral administration of myristic ameliorated glucose tolerance (24-28% decrease in blood glucose levels during glucose tolerance tests) and reduced insulin-responsive blood glucose levels (~20% decrease) in male NSY mice compared with vehicle and groups at 24-30\xa0weeks of age (the age at which the severity of type 2 diabetes is exacerbated in NSY mice). Myristic also attenuated the increase in seen in NSY mice. Furthermore, the fatty increased DGKδ levels (~1.6-fold) in skeletal muscle of NSY mice.These results suggest that the chronic oral administration of myristic improves hyperglycaemia by decreasing insulin-responsive glucose levels and reducing , and that the fatty accounts for the diabetes protective properties of high-fat dairy products. Myristic is a potential candidate for the prevention and treatment of type 2 diabetes mellitus and its related diseases.

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Laundry Detergency of Solid Non-Particulate Soil Using Microemulsion-Based Formulation.

Laundry detergency of solid non-particulate soil on polyester and cotton was investigated using a microemulsion-based formulation, consisting of an anionic extended surfactant (C-4PO-SONa) and sodium mono-and di-methyl naphthalene sulfonate (SMDNS) as the hydrophilic linker, to provide a Winsor Type III microemulsion with an ultralow interfacial tension (IFT). In this work, methyl palmitate ( methyl ester) having a melting point around 30°C, was used as a model solid non-particulate (waxy) soil. A total surfactant concentration of 0.35 wt% of the selected formulation (4:0.65 ratio of C-4PO-SONa:SMDNS) with 5.3 wt% NaCl was able to form a middle phase microemulsion at a high temperature (40°C),which provided the highest oil removal level with the lowest oil redeposition and the lowest IFT, and was much higher than that with a commercial detergent or de-ionized water. Most of the detached oil, whether in liquid or solid state, was in an unsolubilized form. Hence, the dispersion stability of the detached oil droplets or solidified oil particles that resulted from the surfactant adsorption played an important role in the oil redeposition. For an oily detergency, the lower the system IFT, the higher the oil removal whereas for a waxy (non-particulate) soil detergency, the lower the contact angle, the higher the solidified oil removal. For a liquefied oil, the detergency mechanism was roll up and emulsification with dispersion stability, while that for the waxy soil (solid oil) was the detachment by wettability with dispersion stability.

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Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The , relative liver , hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the , relative liver , serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

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Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced , blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

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Exacerbation and Prolongation of Psoriasiform Inflammation in Diabetic Obese Mice: A Synergistic Role of CXCL5 and Endoplasmic Reticulum Stress.

Accumulating evidence suggests that psoriasis is frequently accompanied by metabolic disorders, such as obesity and diabetes. However, the mechanisms underlying the association between increased psoriasis severity and concomitant metabolic syndrome have not been fully clarified. Herein, we show that imiquimod-induced psoriasiform inflammation was exacerbated and prolonged in diabetic obese mice compared to that in control mice, accompanied by remarkably increased lesional expressions of Cxcl5 and Il-1b. Notably, a large number of CXCL5 Ly6G cells infiltrated the dermis and subcutaneous fat tissue of the diabetic obese mice. Most macrophages in the subcutaneous fat tissues of the diabetic obese mice were positive for expression of IL-1β and GRP78/Bip, an endoplasmic reticulum stress marker. Depletion of Ly6G cells and macrophages diminished the imiquimod-induced psoriasiform inflammation. Further, CXCL5 potentiated the secretion of IL-1β from macrophages and , a fatty released from subcutaneous adipocytes, further enhanced IL-1β secretion via endoplasmic reticulum stress induction. Combined with the fact that the serum levels of both CXCL5 and are significantly elevated in patients with metabolic syndrome, our results suggest a role for CXCL5 and endoplasmic reticulum stress in the increase of psoriasis severity of patients with concomitant metabolic syndrome.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

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Association between increased visceral fat area and alterations in plasma fatty profile in overweight subjects: a cross-sectional study.

Visceral fat accumulation in overweight status has been resulted in changes of fatty profiles. The fatty acids\xa0profiles can be altered by fatty desaturase; the activity\xa0of which is highly associated with obesity and other metabolic diseases. We hypothesized that fatty composition, desaturase activity, and accumulation of visceral fat are interrelated. Thus, the aim of this study was to investigate the association between increased visceral fat area and alterations in plasma fatty profile in overweight subjects with different amounts of visceral fat.Healthy overweight subjects (25.0\xa0kg/m\xa0≤\u2009BMI\u2009<\u200930\xa0kg/m,\xa0n=232) were classified into lower (T1), middle (T2), and upper tertiles\xa0(T3) according to L4 visceral fat area (T1: <71.8\xa0cm, T2: 71.8\xa0cm-99.6\xa0cm, T3: >99.6\xa0cm).The T3 group showed higher amounts of cis-10-heptadecenoic and activity of C16 Δ9-desaturase and C18 Δ9-desaturase and lower activity of Δ5-desaturase than the T1 group. Additionally, the T3 group showed higher amounts of saturated fatty acids, myristic , , stearic , monounsaturated fatty acids, palmitoleic , oleic , n-6 polyunsaturated fatty acids, linoleic , dihomo-γ-linolenic , arachidonic , n-3 PUFAs, and docosapentaenoic than the T1 and T2 groups.This study indicates that greater than a certain area (>99.6\xa0cm) of visceral fat is needed to observe altered levels of individual fatty species and desaturase activities. The results suggest that increased activity of C16 Δ9-desaturase and C18 Δ9-desaturase in parallel with decreased Δ5-desaturase activity may be a causative factor in disturbed fatty metabolism.

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A GC-MS untargeted metabolomics analysis in the plasma and liver of rats lacking dipeptidyl-peptidase type IV enzyme activity.

This study was achieved with the aim to find metabolic changes between Fischer rats with different dipeptidyl peptidase-type 4 (DPPIV) expression. The DPPIV is an enzyme expressed in several tissues and is critically involved in the regulation of meal-related insulin secretion in healthy individuals. The metabolic consequences of chronic DPPIV inhibition were analyzed in a surrogate animal model of genetic enzyme deficiency. Hyphenated gas chromatography-mass spectrometry (GC-MS) and multivariate data analysis techniques were used to study the metabolic aqueous fraction profile of 18 plasma and liver samples in two syngeneic rat strains differing in DPPIV activity (DPPIV vs. DPPIV). The hyperglycemic response following oral glucose administration was attenuated in DPPIV rats, as expected. Statistical significant differences between the two strains were observed among the low molecular polar metabolites analyzed from plasma and liver.These included a decrease in malic and glutamine and an increase in pyroglutamic , serine, and alanine in plasma of DPPIV rats. In addition, , L-proline, and ribitol were decreased in the liver of DPPIV strain. Such alterations were compatible with a normal phenotype. These results suggest that long-term exposure to DPPIV inhibitors looks compatible with an overall balanced metabolism.

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Increases Endothelin-1 Expression in Vascular Endothelial Cells through the Induction of Endoplasmic Reticulum Stress and Protein Kinase C Signaling.

We investigated the regulation of endothelin-1 (ET-1) expression in in vivo high-fat diet (HFD)-fed mice and in vitro cultured human aortic endothelial cells (HAECs).Male C57BL/6 mice were fed on standard chow, serum was prepared, and ET-1 levels were analyzed using an ELISA kit. Quantitative PCR was performed using iQ SYBR Green Supermix. Statistical significance was assessed using SPSS, with p < 0.05 considered significant.The serum ET-1 content and endothelial expression of ET-1 mRNA were increased in the HFD-fed mice compared to the chow-fed control mice. Moreover, the mRNA expression of ET-1 was significantly increased in cultured HAECs in response to acute (< 24 h) and chronic (12-16 days) treatments with (PA), one of the most abundant saturated fatty acids in obesity. We found that the induction of ET-1 expression by PA was abolished by pretreating the cells with the endoplasmic reticulum (ER) stress inhibitor 4-phenylbutyric or the protein kinase C (PKC) inhibitor Gö 6850.Our findings demonstrate for the first time that PA increases ET-1 expression in endothelial cells through the induction of ER stress and the activation of PKC, providing novel mechanistic insights into the pathogenesis of obesity-associated hypertension and cardiovascular diseases.© 2018 S. Karger AG, Basel.

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A novel PPARα/γ agonist, propane-2-sulfonic octadec-9-enyl-amide, ameliorates insulin resistance and gluconeogenesis in vivo and vitro.

Peroxisome proliferator-activated receptor alpha/gamma (PPARα/γ) agonists have emerged as important pharmacological agents for improving insulin action. Propane-2-sulfonic octadec-9-enyl-amide (N15) is a novel PPARα/γ dual agonist synthesized in our laboratory. The present study investigates the efficacy and safety of N15 on insulin resistance regulation in high fat diet (HFD)-and streptozotocin (STZ)-induced diabetic mice and in (PA)-induced HepG2 cells. Our results showed that N15 remarkably ameliorated insulin resistance and dyslipidemia in vivo, as well as rectified the glucose consumption and gluconeogenesis in vitro. Moreover, the glucose-lowering effect of N15 was associated with PPARγ mediated up-regulation of hepatic glucose consumption and down-regulation of gluconeogenesis. Meanwhile, N15 exerted advantageous effects on glucose and lipid metabolism without triggering gain and hepatotoxicity in mice. In conclusion, our data demonstrated that by alleviating glucose and lipid abnormalities, N15 could be used as a potential prophylactic and therapeutic agent against type 2 diabetes and related metabolic disorders.Copyright © 2018 Elsevier B.V. All rights reserved.

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Disruption of Lipid Uptake in Astroglia Exacerbates Diet-Induced Obesity.

Neuronal circuits in the brain help to control feeding behavior and systemic metabolism in response to afferent nutrient and hormonal signals. Although astrocytes have historically been assumed to have little relevance for such neuroendocrine control, we investigated whether lipid uptake via lipoprotein lipase (LPL) in astrocytes is required to centrally regulate energy homeostasis. Ex vivo studies with hypothalamus-derived astrocytes showed that LPL expression is upregulated by oleic , whereas it is decreased in response to or triglycerides. Likewise, astrocytic LPL deletion reduced the accumulation of lipid droplets in those glial cells. Consecutive in vivo studies showed that postnatal ablation of LPL in glial fibrillary acidic protein-expressing astrocytes induced exaggerated gain and glucose intolerance in mice exposed to a high-fat diet. Intriguingly, astrocytic LPL deficiency also triggered increased ceramide content in the hypothalamus, which may contribute to hypothalamic insulin resistance. We conclude that hypothalamic LPL functions in astrocytes to ensure appropriately balanced nutrient sensing, ceramide distribution, regulation, and glucose metabolism.© 2017 by the American Diabetes Association.

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Changes of albumin secondary structure after binding. FT-IR spectroscopic study.

Albumin is an universal transport protein. Plasma pool of free fatty acids arising from triglyceride hydrolysis, critical in energy metabolism and etiology of metabolic disorders is transported by albumin. According to various studies albumin has from seven to nine binding sites with diverse affinity to long chain fatty acids. X-ray diffraction crystallography measurements have provided data only for pure human serum albumin or albumin with fully saturated binding sites. These results have shown that amount of \uf061-helices is higher after fatty acids binding. Molecular mechanics simulations suggest that binding of fatty acids in two high-affinity sites leads to major conformational changes in albumin structure. The aim of this research was to investigate albumin secondary structure upon gradually increasing fatty acids to protein mole ratio.Fourier transform infrared spectroscopy was applied to study changes of bovine serum albumin (as an analogue of human serum albumin) \uf061-helical structures after binding in a range of 0-20 : albumin molar ratios representing pure protein, partial, full saturation and excess binding sites capacity.Amount of \uf061-helices was increasing along with the amount of : bovine serum albumin molar ratio and reached maximum value around 2 mol/mol.Our studies confirmed molecular mechanics simulations and crystallographic studies. binding in two high-affinity sites leads to major structural changes, filling another sites only slightly influenced bovine serum albumin secondary structure. The systematic study of fatty acids and albumin interactions, using an experimental model mimicking metabolic disorders, may results in new tools for personalized nanopharmacotherapy.

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Antihyperglycemic, antioxidant activities of tucumã oil (Astrocaryum vulgare) in alloxan-induced diabetic mice, and identification of fatty profile by gas chromatograph: New natural source to treat hyperglycemia.

The aim of the study was to investigate the effect of the oral administration of tucumã oil (Astrocaryum vulgare) on glucose and insulin levels, oxidative status, and pancreatic genotoxic parameters of alloxan-induced diabetic mice. The animals were divided into four groups (n\xa0=\xa06 each): control/water; control/tucumã oil; diabetic/water; diabetic/tucumã oil treated for 14 days with 5.0\xa0mL\xa0kg via oral gavage. Gas chromatograph characterization demonstrated that oleic/elaidic fatty is the most abundant component present in this oil, followed by and stearic fatty acids. Our results demonstrated an increase (p\xa0<\xa00.05) in water and food intake, blood glucose, thiobarbituric reactive species (TBARS) levels, damage index, and frequency of damage; conversely , insulin levels, catalase (CAT) and superoxide dismutase (SOD) activities, and cell viability were decreased in the diabetic/water group compared to the control/water group. The treatment with tucumã oil prevented these alterations in the diabetic/tucumã oil group compared to the diabetic/water group, and restored these parameters near to the control/water group. In summary, our findings demonstrated that treatment with tucumã oil causes a hypoglycemic effect improving insulin levels and antioxidant/oxidant status, and has a protector effect against pancreatic damage induced by oxidative stress in alloxan-induced diabetic mice.Copyright © 2017 Elsevier B.V. All rights reserved.

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-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased , hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid metabolism and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

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Factors affecting human colostrum fatty profile: A case study.

The role of maternal colostrum to infant development has been extensively studied and presented. Among the main factors which contribute to breast milk composition are maternal diet, age and mass index, parity, duration of pregnancy and stage of lactation. This study aims to investigate the potential impact of several factors including demographic (i.e. maternal age and nationality) on the colostrum fatty profile. Colostrum was collected the third day postpartum in a Greek maternity hospital. Certain lipid quality indices and fatty ratios were estimated and results were statistically processed. The main identified fatty acids were (C16:0), oleic (C18:1ω-9), and linoleic (C18:2ω-6) acids. Among fatty acids, saturated fatty acids predominated (47.61%), followed by monounsaturated fatty acids (39.26%), while polyunsaturated fatty acids had the lowest proportion (13.13%). Values of lipid quality indices were within the reported in the literature ranges. Maternal mass index, nationality, age, mode of delivery, gender and fetal percentile were studied in respect to their potential influence on the fatty profile of colostrum fat. Results suggest that colostrum fatty profile was mainly dependent on maternal nationality and age rather than mode of delivery and maternal BMI. Regarding the effect of maternal nationality, significant differences were found for saturated and monounsaturated fatty acids. Of the most interesting findings is that colostrum fat from older (≥35 years) mothers had less saturated fat and more appropriate LQIs values. Finally, a reversed correlation was observed between the customized centile of the infants and the colostrum fat content.

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Beta Palmitate Improves Bone Length and Quality during Catch-Up Growth in Young Rats.

(PA) is the most abundant saturated fatty in human milk, where it is heavily concentrated in the --position (termed beta palmitate, BPA) and as such is conserved in all women, regardless of their diet or ethnicity, indicating its physiological and metabolic importance. We hypothesized that BPA improves the efficiency of nutrition-induced catch up growth as compared to - PA, which is present in vegetable oil. Pre-pubertal male rats were subjected to a 17 days food restriction followed by re-feeding for nine days with PA or BPA-containing diets. We measured bone length, epiphyseal growth plate height (EGP, histology), bone quality (micro-CT and 3-point bending assay), and gene expression (Affymetrix). The BPA-containing diet improved most growth parameters: humeri length and EGP height were greater in the BPA-fed animals. Further analysis of the EGP revealed that the hypertrophic zone was significantly higher in the BPA group. In addition, Affymetrix analysis revealed that the diet affected the expression of several genes in the liver and EGP. Despite the very subtle difference between the diets and the short re-feeding period, we found a small but significant improvement in most growth parameters in the BPA-fed rats. This pre-clinical study may have important implications, especially for children with growth disorders and children with special nutritional needs.

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Bioconversion of cheese whey permeate into fungal oil by .

Oleaginous fungi are efficient tools to convert agricultural waste streams into valuable components. The filamentous fungus was cultivated in whey permeate, a byproduct from cheese production, to produce an oil-rich fungal biomass. Response surface methodology was used to optimize the fermentation conditions such as pH and temperature for increased biomass yield and lipid accumulation. Quantification and characterization of the fungal biomass oil was conducted.Upstream lactose hydrolysis of the whey permeate increased the biomass yield from 2.4 to 7.8 (g dry biomass/L) compared to that of non-hydrolyzed whey permeate. The combination of low pH (4.5) and pasteurization minimized microbial competition, thus favoring fungal growth. A central composite rotatable design was used to evaluate the effects of temperature (22.4-33.6\xa0°C) and a lower pH range (3.6-4.7) on biomass yield and composition. The highest biomass yield and oil content was observed at high temperature (33.6\xa0°C), while the pH range evaluated had a less pronounced effect. The predictive model was validated at the optimal conditions of 33.6\xa0°C and pH\xa04.5. The fungal biomass yield plateaued at 9\xa0g dry cell per liter, while the oil content and lipid yield reached a maximum of 24% dry biomass and 2.20\xa0g/L, respectively, at 168\xa0h. Triacylglycerides were the major lipid class (92%), which contained predominantly oleic (41%), (23%), linoleic (11%), and γ-linolenic (9%).This study provided an alternative way of valorization of cheese whey permeate by using it as a substrate for the production of value-added compounds by fungal fermentation. The fatty profile indicates the suitability of oil as a potential feedstock for biofuel production and nutraceutical applications.

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Fatty Profile, Total Carotenoids, and Free Radical-Scavenging from the Lipophilic Fractions of 12 Native Mexican Avocado Accessions.

Mexico is the world\'s largest producer, exporter, and consumer of avocados. "Hass" avocado is the most commercialized cultivar, while the native Mexican avocado varieties have been displaced. Thus, studies regarding their chemical and nutraceutical characterization are scarce. We studied the total lipid content, fatty profile, carotenoid content, and free radical-scavenging activity of the fruit pulp from 12 accessions of the native Mexican avocado (Persea americana var. drymifolia). The results show that the chemical and nutraceutical characteristics depend on the accession type. The total lipid content ranged from 13.22 to 23.41%. The major saturated fatty in all the avocado accessions was (15.54-22.68%). Monounsaturated fatty acids, like oleic (56.58-74.19%), linoleic (5.62-16.85%) and linolenic (0.63-2.85%) acids were the most abundant unsaturated fatty acids in all the native Mexican avocado accessions. The total carotenoid content (1.57 to 7.83\xa0mg\xa0eq. of β-carotene 100\xa0g- fresh ) and the free radical-scavenging activity (7.58-88.62 mMol trolox equivalent 100\xa0g fresh ) also varied significantly (p\u2009<\u20090.05) among accessions. Native Mexican avocados have a great nutraceutical potential due to their high content of omega-9, omega-6, and omega-3 fatty acids and carotenoids. These compounds have been reported to display antioxidant activities and protect against cardiovascular diseases.

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Isolation and Selection of Microalgal Strains from Natural Water Sources in Viet Nam with Potential for Edible Oil Production.

Industrial vegetable oil production in Viet Nam depends on oil seeds and crude plant oils that are currently more than 90% imported. As the first step in investigating the feasibility of using microalgae to provide Viet Nam with a domestic source of oil for food and edible oil industries, fifty lipid-producing microalgae were isolated and characterized. The microalgae were isolated from water sources ranging from freshwater to brackish and marine waters from a wide geographic distribution in Viet Nam. Initial analyses showed that 20 of the 50 strains had good growth rates, produced high biomass and had high lipid content, ranging up to 50% of dry biomass. 18S rRNA gene sequence analyses of the 50 strains showed a great diversity in this assemblage of microalgae, comprising at least 38 species and representatives of 25 genera, , , , , , , , , , , , , , , , , , , , , , , , and Some of the species are closely related to well-known lipid producers such as , but some other strains are not closely related to the strains found in public sequence databases and likely represent new species. Analysis of oil quality showed that fatty profiles of the microalgal strains were very diverse and strain-dependent. Fatty acids in the microalgal oils comprised saturated fatty acids (SFAs), poly-unsaturated fatty acids (PUFAs), and mono-unsaturated fatty acids (MUFAs). The main SFA was . MUFAs and PUFAs were dominated by oleic , and linoleic and linolenic acids, respectively. Some strains were especially rich in the essential fatty α-linolenic (ALA), which comprised more than 20% of the fatty acids in these strains. Other strains had fatty compositions similar to that of palm oil. Several strains have been selected on the basis of their suitable fatty profiles and high lipid content for further chemical and physical characterization, toxicity and organoleptic tests of their oils, and for scale-up.

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